muramidase and Inflammation

The gastrointestinal tract is a complex environment in which the host immune system interacts with a diverse array of microorganisms, both symbiotic and pathogenic. As such, mobilizing a rapid and appropriate antimicrobial response depending on the nature of each stimulus is crucial for maintaining the balance between homeostasis and inflammation in the gut. Here we focus on the mechanisms by which intestinal antimicrobial peptides regulate microbial communities during dysbiosis and infection. We also discuss classes of bacterial peptides that contribute to reducing enteric pathogen outgrowth. This review aims to provide a comprehensive overview on the interplay of diverse antimicrobial responses with enteric pathogens and the gut microbiota.

Topics: Animals; Bacteriocins; Cathelicidins; Defensins; Dysbiosis; Gastrointestinal Microbiome; Gastrointestinal Tract; Gene Expression; Humans; Immunity, Mucosal; Inflammation; Intestinal Mucosa; Lipocalin-2; Muramidase; Symbiosis

The mucosa of the esophagus, the stomach, the small intestine, the large intestine and rectum are unremittingly challenged by adverse micro-environmental factors, such as ingested pathogenic and non-pathogenic bacteria, and harsh secretions with digestive properties with disparate pH, as well as bacteria and secretions from upstream GI organs. Despite the apparently inauspicious mixture of secretions and bacteria, the normal GI mucosa retains a healthy state of cell renewal. To by-pass the tough microenvironment, the epithelia of the GI react by speeding-up cell exfoliation, by increasing peristalsis, eliminating bacteria through secretion of plasma cell-immunoglobulins and by increasing production of natural antibacterial enzymes (lysozyme) and host defense peptides (defensin-5). Lysozyme was recently found up-regulated in Barrett's esophagitis, in chronic gastritis, in gluten-induced atrophic duodenitis (celiac disease), in collagenous colitis, in lymphocytic colitis and in Crohn's colitis. This up-regulation is a response directed towards the special types of bacteria thriving in the microenvironment in each of the aforementioned clinical inflammatory maladies. The purpose of that up-regulation is to protect the mucosa affected by the ongoing chronic inflammation. Bacterial antibiotic resistance continues to exhaust our supply of effective antibiotics. The future challenge is how to solve the increasing menace of bacterial resistance to anti-bacterial drugs. Further research on natural anti-bacterial enzymes such as lysozyme, appears mandatory.

Topics: Barrett Esophagus; Celiac Disease; Colitis, Collagenous; Colitis, Lymphocytic; Colitis, Ulcerative; Crohn Disease; Gastritis; Gastrointestinal Diseases; Humans; Inflammation; Muramidase

Antimicrobial proteins (AMP) are endogenous, gene-encoded proteins, which are able to kill bacteria, fungi and viruses at micro- and nanomolar concentrations. The constitutive as well as inducible production of AMP provides a rapid first-line of defense against invading microorganisms. The significance of such ancient defense system is reflected by the wide distribution of AMP in the plant and animal kingdom. There is increasing evidence that AMP may play an important role in several infectious and inflammatory diseases such as atopic dermatitis, cystic fibrosis and Crohn's disease. In this review we aim to provide a short overview about the role of antimicrobial proteins in human diseases. In addition, the use and selective induction of AMP for the development of novel potential therapeutic strategies are addressed. The benefits and possible restrictions of AMP utilization as a new class of antibiotic compounds are discussed.

Topics: Animals; Anti-Infective Agents; Anti-Inflammatory Agents, Non-Steroidal; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Defensins; Gastrointestinal Diseases; Humans; Infections; Inflammation; Muramidase; Phagocytes; Proteins; Skin Diseases

Several natural components abundant in the fluid phase of human breast-milk have been shown to be inhibitors of complement activation in vitro, particularly the classical pathway. These include lysozyme, lactoferrin, lactalbumin alpha and other ligand chelators, complement regulator proteins and other specific soluble inhibitors of complement activation. Their physiological significance probably resides in their ability to restrict in vivo complement activation to specialized (compartmentalized) sites on the cellular membrane structures in human milk, represented by the abundant surface area of the milk fat globule membranes. This would serve to prevent inflammatory-induced tissue damage of the delicate immature gastrointestinal tract of the newborn as well as the mammary gland itself. A number of recognized and potential inhibitors of complement activity in human milk and other biological fluids are hereby reviewed, with a proposal of their physiological significance.

Topics: Animals; Complement Inactivator Proteins; Female; Humans; Immunoglobulins; Infant, Newborn; Inflammation; Lactalbumin; Lactoferrin; Mammary Glands, Animal; Milk, Human; Muramidase

Histo-monocytes are cells playing an important role in host defense reaction and purification of the organism. These cells belong to a new system of highly phagocytic mononuclear cells termed "mononuclear phagocyte system". This system includes the promonocytes and their precursors in the bone marrow, the monocytes in the peripheral blood and the different types of histiocytes and macrophages particularly Kupffer cells. Histiocytes can also transform into epithelioid cells and giant cells. The inclusion of all these cells in this system is based on similarities in the morphology, function, origin and kinetics of the phagocytes. The most important morphologic characteristics of these cells are described. It is also shown that the mononuclear phagocytic system is a secretory system. The different types of products are described. Recently, so-called accessory cells of the immunity were described, comprising dendritic reticular cells from the follicles and interdigitating reticular cells from the deep cortical zone of the lymph node. Those cells belong both probably to the system. The cells of this system are engaged in the clearance of multiple organs, in the inflammatory process, in the immune response and in the immune surveillance against tumor.

Topics: Animals; Antibody Formation; Blood Coagulation; Complement System Proteins; Hematopoiesis; Histiocytes; Humans; Hydrolases; Immunity, Cellular; Inflammation; Lipid Metabolism; Monocytes; Monokines; Muramidase; Peptide Hydrolases; Phagocytes; Phagocytosis; Proteins; Stem Cells

Topics: Antigen-Antibody Complex; B-Lymphocytes; Humans; Immunity, Innate; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Inflammation; Leukocyte Count; Muramidase; Opsonin Proteins; Phagocytosis; Postoperative Period; Suppuration; T-Lymphocytes

During the intrauterine period, the human immunologic system develops through a complex but orderly series of events. The functional capacity of the system remains incomplete, not only during prenatal life but also through much of infancy. Many of the factors not produced by the fetus or infant are provided by the mother. Systemic immunity is augmented by specific IgG antibodies from the placenta and mucosal immunity by a wide array of defense agents from human milk including sIgA antibodies, lactoferrin, lysozyme, other soluble factors with antimicrobial properties, and specifically adapted leukocytes. It appears that the defense of the infant and the maternal contribution to that defense are geared to protect principally by noninflammatory mechanisms. Although much has been discovered about the ontogeny of the human immunologic system and the maternal contributions to this immunity, much remains to be learned about the molecular controls of the system, the fate of the transported maternal factors, feedback mechanisms between the immunologic systems of the mother and infant and the precise effects of maternal factors upon the infant. Answers to these questions may lead to the development of immunizing agents which are better suited to the infant, mucosal immunogens fashioned to stimulate the production of protective SIgA antibodies in human milk, the provision of defense factors for serious infections in young infants, and ways to enhance the maturation of the immunologic system of the infant when that is desirable.

Topics: Antibody Formation; B-Lymphocytes; Complement System Proteins; Female; Fetus; Humans; Immune System; Immunity, Cellular; Immunity, Maternally-Acquired; Immunoglobulin A, Secretory; Infant, Newborn; Inflammation; Lactoferrin; Leukocytes; Milk, Human; Muramidase; Phagocytes; Pregnancy; T-Lymphocytes

Neutrophilic polymorphonuclear leukocytes (NPNL) are an important part of the protection system of higher animals against infection and related with specific immune response taking part in immediate and delayed type hypersensitivity reactions and affecting some parameters of mononuclear phagocytes and lymphocytes. Among bactericidal mechanisms of NPNL, neutral proteases and nonenzymatic cation proteins are particularly important. NPNL have fine autoregulation mechanisms among which of special importance is the synthesis of biologically active substances: leukotrienes, including a slow acting anaphylaxis substance, prostaglandins, and prostacyclins. These metabolites of arachidonic acid affect chemotropism of NPNL, phagocytosis activity, and secretion processes. The real activity of NPNL bactericidal mechanisms is determined by the following factors: (1) the state of genetically programmed structural-functional parameters common to NPNL, (2) the state of autoregulation mechanisms of NPNL function, (3) the stage of the infectious process, (4) NPNL--macrophage-lymphocyte system interaction, (5) regulatory effect of the neurohumoral and endocrine systems, (6) serum factors of nonspecific resistance, (7) characteristics of the causative agent of infection.

Topics: Animals; Chemotaxis, Leukocyte; Fatty Acids, Unsaturated; Guinea Pigs; Humans; Immunity, Innate; Inflammation; Macrophage Activation; Macrophages; Muramidase; Neutrophils; Peptide Hydrolases; Rabbits

Topics: Bacteria; Burns; Cell Wall; Colitis, Ulcerative; Crohn Disease; DNA, Bacterial; Humans; Inflammation; Leukemia; Muramidase; RNA, Bacterial

Phagocytosis begins with exocytosis--"extrarapid" discharge of bactericidal proteins and factors of permeability into the extracellular medium. A viewpoint was put forward on an "avalanch-like" character of the outcome of cationic proteins from leukocyte granules in inflammation and their participation in formation of a nonphagocytic type of resistance. In phagocytosis bacteria perish due to the myeloperoxidase system, lysozyme, lactoferin and nonenzymic cationic proteins. Hereditary deficit of the above-mentioned substances leads to intraleukocytic microbicidal insufficiency, a drastic decrease in the nonspecific resistance of the organism and to development of fatal granulomatous disease, and to other forms of pathology associated with genetic defects of the bactericidal systems of leukocytes.

Topics: Blood Bactericidal Activity; Blood Proteins; Cell Membrane Permeability; Cytoplasmic Granules; Exocytosis; Glucosephosphate Dehydrogenase Deficiency; Histones; Humans; Immunity; Immunologic Deficiency Syndromes; Inflammation; Lactoferrin; Microscopy, Phase-Contrast; Muramidase; Neutrophils; Peroxidase; Phagocytosis

Topics: Actins; Animals; Bone Marrow; Bone Marrow Cells; Cell Differentiation; Cell Membrane; Cell Nucleus; Cytoplasmic Granules; Endoplasmic Reticulum; Golgi Apparatus; Hematologic Diseases; Hematopoiesis; Histocytochemistry; Humans; Hydrolases; Inflammation; Leukemia; Muramidase; Myosins; Neutrophils; Peroxidase; Peroxidases

Topics: Animals; Antibody Formation; Classification; Colony-Stimulating Factors; Complement System Proteins; DNA Replication; Humans; Immunity; Inflammation; Interferons; Lymphocytes; Lysosomes; Macrophages; Microbial Collagenase; Monocytes; Muramidase; Pancreatic Elastase; Phagocytes; Plasminogen Activators; Pyrogens; Toxins, Biological; Wound Healing

Although it is clear that macrophages are always present at sites of chronic inflammation their contribution to the evolution of these lesions is not well understood. In vitro studies have shown that macrophages secrete a variety of products on exposure to different stimuli. These include hydrolytic enzymes, active at acid or neutral pH, with known capacity for degrading tissue constituents. Lysosomal acid hydrolases are released from viable cells over a prolonged period of time by various agents known to cause, or are associated with, chronic inflammation. These agents may be nonimmunogenic substances, such as carrageenan and asbestos, which interact directly with macrophages or alternatively the products of immune reactions involving either B or T lymphocytes. These lymphocyte products include immune complexes of certain composition and the secreted products of T lymphocytes stimulated by nonspecific mitogens or specific antigens. In marked contrast biologically inactive substances such as latex particles or digestible substrates such as erythrocytes do not induce the selective release of acid hydrolases from macrophages. It is clear that alghough macrophages secrete abundant amounts of neutral proteinases under certain conditions this release does not occur necessarily during the release of acid hydrolases induced by inflammatory agents. The role played by acid and neutral hydrolases secreted by macrophages during the various stages of chronic inflammatory responses remains to be clarified.

Topics: Animals; Antigen-Antibody Complex; Asbestos; Carrageenan; Cell Wall; Cells, Cultured; Chronic Disease; Guinea Pigs; Humans; Hydrolases; Inflammation; Lysosomes; Macrophages; Mice; Microbial Collagenase; Muramidase; Plasminogen Activators; Prostaglandins; Rats; Silicon Dioxide; Stimulation, Chemical; Streptococcus; T-Lymphocytes; Zymosan

Topics: Antibody Formation; Antibody Specificity; Antigen-Antibody Reactions; B-Lymphocytes; Cilia; Communicable Diseases; Complement System Proteins; Epithelium; Epitopes; Humans; Hypersensitivity, Delayed; Immunity; Immunity, Cellular; Immunoglobulin A; Immunologic Memory; Inflammation; Macrophages; Mononuclear Phagocyte System; Mucous Membrane; Muramidase; Neutrophils; Phagocytosis; T-Lymphocytes

Topics: Antibodies, Heterophile; Antibody Formation; Blood Coagulation; Cell Migration Inhibition; Cytotoxicity Tests, Immunologic; Graft Rejection; Humans; Immune Adherence Reaction; Immunity, Cellular; Inflammation; Kidney Transplantation; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Muramidase; Proteinuria; Transplantation, Homologous

Topics: Blood Proteins; Cytoplasm; Cytoplasmic Granules; Glucose; In Vitro Techniques; Inflammation; Lactates; Lactoferrin; Lysosomes; Muramidase; NADH, NADPH Oxidoreductases; Peroxidase; Phagocytes; Phagocytosis

Topics: Anti-Bacterial Agents; Bacteriuria; Blood; C-Reactive Protein; Cerebrospinal Fluid; Complement System Proteins; Delivery, Obstetric; Female; Fetal Diseases; Humans; Immunoglobulins; Infant Care; Infant, Newborn; Infant, Newborn, Diseases; Infection Control; Infections; Inflammation; Leukocyte Count; Maternal-Fetal Exchange; Muramidase; Phagocytosis; Pregnancy; Sex Factors; Sterilization

Topics: Alkaline Phosphatase; Antigen-Antibody Complex; Blood Bactericidal Activity; Chemotaxis; Complement System Proteins; Cytoplasmic Granules; Deoxyribonucleases; Erythrocytes; Escherichia coli; Glucuronidase; Granuloma; Humans; Inflammation; Leukocytes; Lysosomes; Macrophages; Monocytes; Muramidase; Neutrophils; Opsonin Proteins; Organoids; Phagocytosis; Ribonucleases

Topics: Animals; Anti-Bacterial Agents; Anti-Infective Agents; Anti-Inflammatory Agents; Bacteria; Drug Synergism; Female; Fever; Genital Diseases, Female; Guinea Pigs; Haplorhini; Humans; Infections; Inflammation; Mice; Muramidase; Penicillins; Peritoneal Diseases; Phagocytosis; Pneumococcal Infections; Rabbits; Rats; Staphylococcal Infections; Urologic Diseases; Virus Diseases; Viruses

Topics: Biomarkers; Humans; Immunoglobulin A, Secretory; Inflammation; Lasers, Semiconductor; Low-Level Light Therapy; Muramidase; X-Rays

To evaluate gingival inflammation from fixed-dose combinations of vitamin C, vitamin E, lysozyme and carbazochrome (CELC) in the treatment of chronic periodontitis following scaling and root planing.. One hundred patients were randomly assigned to receive CELC (test) or placebo (control) for the first 4 weeks at a 1:1 ratio, and both groups received CELC for the remaining 4 weeks. Primary outcome was the mean change in the gingival index (GI) after 4 weeks. Secondary outcomes included mean change in GI after 8 weeks and plaque index, probing depth, clinical attachment level, and VAS at 4 weeks and 8 weeks.. Ninety-three patients completed the study. The GI in the test group significantly decreased after 4 weeks (p < 0.001) and 8 weeks (p < 0.001). The mean change from baseline in GI significantly decreased in the test group compared to the control group after 4 weeks (p = 0.015). In the GEE model adjusting for age, gender and visits, the test group showed 2.5 times GI improvement compared to the control group (p = 0.022).. Within the study, CELC showed a significant reduction in gingival inflammation compared with a placebo. Other parameters, however, were similar between groups.. KCT0001366 (Clinical Research Information Service, Republic of Korea) and 29 Jan 2015, retrospectively registered.

Topics: Adrenochrome; Anti-Bacterial Agents; Ascorbic Acid; Chronic Periodontitis; Dental Plaque Index; Dental Scaling; Double-Blind Method; Drug Therapy, Combination; Gingival Crevicular Fluid; Humans; Inflammation; Muramidase; Republic of Korea; Retrospective Studies; Root Planing; Vitamin E

The host inflammatory response to particulate wear debris has been implicated as a principal cause of osteolysis and aseptic loosening following total joint arthroplasty. While it has long been assumed that this inflammatory response is mediated solely by a chronic process, there has been evidence to suggest that an acute response to particulate debris may be important in initiating the chronic response. We studied the in vitro and in vivo acute inflammatory responses mediated by polymorphonuclear leukocytes (PMNs) to both retrieved particulate from a catastrophically failed uncemented metal-backed acetabular component and to commercially pure particulate (polyethylene, cobalt-chrome, and titanium). Isolated, nonactivated human PMNs in vitro exhibited both a dose- and time-dependent degranulation response to opsonized particulate debris, as evidenced by release of both specific (increased lysozyme activity) and azurophilic (increased beta-glucuronidase activity) granule contents. In the rat subcutaneous pouch model in vivo, PMNs were recruited within 3-6 h after exposure to particulate debris and were noted to phagocytize particulate and subsequently degranulate, as evidenced by increased beta-glucuronidase and PMN-specific myeloperoxidase (azurophilic granule enzymes) activities. This response peaked within the first 6 h and gradually declined by 24 h. The results of this study demonstrate the presence of an acute inflammatory response mediated by PMNs both in vitro and in vivo to particulate debris, which may be important in the sequence of events that lead to the macrophage-dominated chronic inflammatory process culminating in osteolysis and aseptic loosening of total joint arthroplasties.

Topics: Animals; Arthroplasty, Replacement, Hip; Cell Degranulation; Centrifugation, Density Gradient; Chromium Alloys; Glucuronidase; Humans; Inflammation; L-Lactate Dehydrogenase; Microscopy, Electron, Scanning; Muramidase; Neutrophil Activation; Neutrophils; Opsonin Proteins; Polyethylene; Prosthesis Failure; Rats; Titanium

Chronic bronchitis is associated with airways obstruction and inflammation. In order to determine whether aerosolized beclomethasone can modulate airway inflammation and diminish airway obstruction, subjects with chronic bronchitis performed spirometry and underwent bronchoalveolar lavage (BAL) before and after receiving 6 wk of therapy (five puffs four times a day) with either aerosolized beclomethasone (n = 20) or placebo (n = 10) in a double-blinded, randomized fashion. All subjects received aerosolized albuterol before each use of the study medications. Before BAL, the airways were visually assessed for the appearance of inflammation and assigned a score, the bronchitis index. BAL was performed by instilling five 20-ml aliquots of saline into each of three sites and pooling and separately analyzing the returns from the first aliquots to yield a "bronchial sample." The bronchial lavages were repeated in an additional three sites to increase the volume of fluid available for analysis. The fluid was prepared for cytologic examination by cytocentrifugation. Albumin (as a measure of epithelium permeability) and lactoferrin and lysozyme (as measures of serous cell activity) were measured in unconcentrated BAL fluid by enzyme-linked immunosorbent assay, and concentrations in epithelial lining fluid were estimated using urea as an internal marker for dilution. After treatment, the beclomethasone group, but not the placebo group, showed improvement in FVC (p = 0.02), FEV1 (p = 0.002), and 25 to 75% forced expiratory flow (p = 0.006). Associated with the improvement in spirometry, the bronchitis index fell (13.5 +/- 1.0 versus 10.75 +/- 1.1, p = 0.02) in the beclomethasone-treated group, but not the placebo-treated group.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Administration, Inhalation; Adult; Aerosols; Airway Obstruction; Albumins; Beclomethasone; Blood Gas Analysis; Bronchitis; Bronchoalveolar Lavage Fluid; Bronchoscopy; Chronic Disease; Double-Blind Method; Female; Forced Expiratory Volume; Humans; Inflammation; Lactoferrin; Male; Middle Aged; Muramidase; Smoking; Transferrin; Vital Capacity

This study shows that the supramolecular arrangement of proteins in nanoparticle structures predicts nanoparticle accumulation in neutrophils in acute lung inflammation (ALI). We observed homing to inflamed lungs for a variety of nanoparticles with agglutinated protein (NAPs), defined by arrangement of protein in or on the nanoparticles via hydrophobic interactions, crosslinking and electrostatic interactions. Nanoparticles with symmetric protein arrangement (for example, viral capsids) had no selectivity for inflamed lungs. Flow cytometry and immunohistochemistry showed NAPs have tropism for pulmonary neutrophils. Protein-conjugated liposomes were engineered to recapitulate NAP tropism for pulmonary neutrophils. NAP uptake in neutrophils was shown to depend on complement opsonization. We demonstrate diagnostic imaging of ALI with NAPs; show NAP tropism for inflamed human donor lungs; and show that NAPs can remediate pulmonary oedema in ALI. This work demonstrates that structure-dependent tropism for neutrophils drives NAPs to inflamed lungs and shows NAPs can detect and treat ALI.

Topics: Acute Disease; Agglutination; Animals; Antibodies; Cross-Linking Reagents; Dextrans; Humans; Inflammation; Lipopolysaccharides; Liposomes; Lung; Male; Mice, Inbred C57BL; Muramidase; Nanoparticles; Neutrophils; Opsonin Proteins; Proteins; Static Electricity; Tissue Distribution; Tomography, Emission-Computed, Single-Photon; Tomography, X-Ray Computed

Protein aggregation can affect the stability and function of proteins, and may lead to developing diseases, but reports on the in vivo effect of aggregates are scarce. In the current study, the effect of phenylalanine (Phe) and indole presence was first investigated on the structure and stability of human lysozyme (HLZ) and its aggregation under in vitro condition. Tm measurements, circular dichroism and spectrofluorimetric spectra, as well as and transmission electron microscopy (TEM) were performed in this stage. In the next step, pathogenicity of HLZ amorphous aggregates formed in presence or absence of the additives was investigated in vivo, by subcutaneous injection to adult male Wistar rats. Resulting inflamed tissues were studied by hematoxylin and eosin (HE), Congo red and Sudan black staining. Serum levels of liver enzymes (Alanine Aminotransferase (ALT) and Aspartate Aminotransferase (AST)), specific inflammatory cytokines (Tumor Necrosis Factor Alpha (TNF-α) and Interleukin 6 (IL-6)) as well as glucose, cholesterol, and triglyceride levels were measured. Amorphous aggregates of HLZ caused inflammation and affected the number of fat cells, macrophages, cytokines, liver enzymes and glucose. Indole, that increases amorphous aggregates amount as shown with CD, fluorescence, and TEM experiments, leads into more severe inflammation. In presence of Phe, (which stabilizes HLZ structure) a markedly milder inflammatory state is observed in histological results and no increase could be detected in the inflammation-related parameters. In conclusion, amorphous aggregates of HLZ may be pathogenic in vivo, and presence of anti-aggregation compounds (such as Phe) can be effective in diminishing their deleterious manifestations.

Topics: Animals; Cytokines; Glucose; Humans; Indoles; Inflammation; Liver; Male; Muramidase; Phenylalanine; Rats; Rats, Wistar; Virulence

The purpose of this study was to investigate the effects of lysozyme, an antimicrobial enzyme found in tears that protects the eye against pathogens, on pseudotyped severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection through corneal epithelial cells.. The expression of the angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease (TMPRSS2) in human corneal epithelial cells (HCECs) was measured by RT-PCR and Western blotting. The altered expression of the pro-inflammatory molecules induced by spike protein and lysozyme was analyzed by RT-PCR. Cell toxicity was tested by CCK8 assay. The cell entry of SAR-CoV-2 in HCECs and primary rabbit corneal epithelial cells (RbCECs) was detected by luciferase assay.. ACE2 and TMPRSS2 were highly expressed in HCECs. The spike proteins of SARS-CoV-2 stimulated a robust inflammatory response in HCECs, characterized by increased secretion of pro-inflammatory molecules, including IL-6, TNF-α, iNOS, and MCP-1, and pretreatment with lysozyme in HCECs markedly decreased the production of proinflammatory molecules induced by spike proteins. In addition, the inflammatory cytokine TNF-α enhanced the entry of SARS-CoV-2 into HCECs, which can be mitigated by pretreatment with lysozyme.. In this study, we analyzed the susceptibility of human corneal epithelial cells to SARS-CoV-2 infection and suggested the protective effects of lysozyme on SARS-CoV-2 infection.

Topics: Angiotensin-Converting Enzyme 2; Animals; Antiviral Agents; COVID-19; Epithelial Cells; Humans; Inflammation; Muramidase; Peptidyl-Dipeptidase A; Rabbits; SARS-CoV-2; Spike Glycoprotein, Coronavirus; Tumor Necrosis Factor-alpha

Nuclear factor-κB (NF-κB) plays a role as a rheostatic transcription factor in regulating intestinal inflammation, and its disruption or constitutive activation leads to inflammation and injury. However, the molecular mechanisms of NF-κB regulation remain largely unknown. In this study, the NF-κB-regulated host defenses against pathogen infections and facilitation of IL17 expression during stimulation with different bacteria were investigated. Intestinal inflammation was induced by dextran sulfate sodium, and NF-κB activity was inhibited in an intestinal injury model. Mannose receptor C type, ABF1/2, serpin B13, lysozyme, and β-arrestin were significantly controlled by NF-κB in the inflamed intestinal tissue. High levels of NF-κB activation resulted in less pervasive intestinal damage and the maintenance of intestinal barrier integrity. Intestinal injury robustly increased the expression of IL17. NF-κB activation was enhanced by IL17 deficiency in the intestinal injury model. IL17 inhibition aggravated intestinal inflammation, leading to loss of epithelial architecture and the infiltration of inflammatory cells. These data suggest that NF-κB and IL17 play key mediator roles in the maintenance of gut epithelial integrity and immune homeostasis.

Topics: Animals; Artemia; beta-Arrestins; Dextran Sulfate; Inflammation; Intestinal Mucosa; Muramidase; NF-kappa B; Serpins

Several proteins of the innate immune system are known to be deregulated with insulin resistance. We here aimed to investigate the relationship among circulating lysozyme (both plasma concentration and activity) and obesity-associated metabolic disturbances.. Plasma lysozyme concentration was determined cross-sectionally in a discovery (Cohort 1, n = 137) and in a replication cohort (Cohort 2, n = 181), in which plasma lysozyme activity was also analyzed. Plasma lysozyme was also evaluated longitudinally in participants from the replication cohort (n = 93). Leukocyte lysozyme expression (LYZ mRNA) were also investigated in an independent cohort (Cohort 3, n = 76), and adipose tissue (AT) LYZ mRNA (n = 25) and plasma peptidoglycan levels (n = 61) in subcohorts from discovery cohort.. Translocation of peptidoglycan (as inferred from its increased circulating levels) was linked to plasma lysozyme, hyperinsulinemia and dyslipidemia in obese subjects. In both discovery and replication cohorts, plasma lysozyme levels and activity were significantly increased in obesity in direct association with obesity-associated metabolic disturbances and inflammatory parameters, being circulating lysozyme negatively correlated with fasting glucose, HbA1c and insulin resistance (HOMA-IR) in obese subjects. Of note, total cholesterol (p < 0.0001) and LDL cholesterol (p = 0.003) contributed independently to age-, gender- and BMI adjusted plasma lysozyme activity. Longitudinally, changes in HbA1c levels and serum LDL cholesterol were negatively associated with circulating lysozyme antimicrobial activity. On the contrary, the change in glucose infusion rate during the clamp (insulin sensitivity) was positively associated with lysozyme concentration.. Increased plasma lysozyme levels and activity are found in obese subjects. The longitudinal findings suggest that plasma lysozyme might be protective on the development of obesity-associated metabolic disturbances.

Topics: Adipose Tissue; Adult; Blood Glucose; Cohort Studies; Dyslipidemias; Female; Glucose Intolerance; Humans; Immune System; Inflammation; Insulin Resistance; Longitudinal Studies; Male; Middle Aged; Muramidase; Obesity; Peptidoglycan

Muramidases constitute a superfamily of enzymes that hydrolyse peptidoglycan (PGN) from bacterial cell walls. Recently, a fungal muramidase derived from Acremonium alcalophilum has been shown to increase broiler performance when added as a feed additive. However, the underlying mechanisms of action are not yet identified. Here, we investigated the hypothesis that this muramidase can cleave PGN to muramyl dipeptide (MDP), activating nucleotide-binding oligomerisation domain-containing protein 2 (NOD2) receptors in eukaryotic cells, potentially inducing anti-inflammatory host responses. Using Micrococcus luteus as a test bacterium, it was shown that muramidase from A. alcalophilum did not display antimicrobial activity, while it could cleave fluorescently labelled PGN. It was shown that the muramidase could degrade PGN down to its minimal bioactive structure MDP by using UPLC-MS/MS. Using HEK-Blue™-hNOD2 reporter cells, it was shown that the muramidase-treated PGN degradation mixture could activate NOD2. Muramidase supplementation to broiler feed increased the duodenal goblet cell and intraepithelial lymphocyte abundance while reducing duodenal wall CD3+ T lymphocyte levels. Muramidase supplementation to broiler feed only had moderate effects on the duodenal, ileal and caecal microbiome. It was shown that the newly discovered muramidase hydrolysed PGN, resulting in MDP that activates NOD2, potentially steering the host response for improved intestinal health.

Topics: Acetylmuramyl-Alanyl-Isoglutamine; Animal Nutritional Physiological Phenomena; Animals; Bacteria; Cell Wall; Cells, Cultured; Chickens; Chromatography, Liquid; Duodenum; Inflammation; Muramidase; Nod2 Signaling Adaptor Protein; Peptidoglycan; Tandem Mass Spectrometry

Chronic systemic low-level inflammation in metabolic disease is known to affect adipose tissue biology. Lysozyme (LYZ) is a major innate immune protein but its role in adipose tissue has not been investigated. Here, we aimed to investigate LYZ in human and rodents fat depots, and its possible role in obesity-associated adipose tissue dysfunction. LYZ mRNA and protein were identified to be highly expressed in adipose tissue from subjects with obesity and linked to systemic chronic-low grade inflammation, adipose tissue inflammation and metabolic disturbances, including hyperglycemia, dyslipidemia and decreased markers of adipose tissue adipogenesis. These findings were confirmed in experimental models after a high-fat diet in mice and rats and also in ob/ob mice. Importantly, specific inguinal and perigonadal white adipose tissue lysozyme (Lyz2) gene knockdown in high-fat diet-fed mice resulted in improved adipose tissue inflammation in parallel to reduced lysozyme activity. Of note, Lyz2 gene knockdown restored adipogenesis and reduced weight gain in this model. In conclusion, altogether these observations point to lysozyme as a new actor in obesity-associated adipose tissue dysfunction. The therapeutic targeting of lysozyme production might contribute to improve adipose tissue metabolic homeostasis.

Topics: Adipogenesis; Adipose Tissue; Animals; Diet, High-Fat; Gene Knockdown Techniques; Inflammation; Male; Mice; Mice, Inbred C57BL; Muramidase; Obesity; Rats, Wistar

In deep burns, wound contraction and hypertrophic scar formation can generate functional derangement and debilitation of the affected part. In order to improve the quality of healing in deep second-degree burns, we developed a new treatment in a preclinical model using nanostructured membranes seeded with mesenchymal stem cells (MSCs).. Membranes were obtained by reconstitution of bacterial cellulose (reconstituted membrane [RM]) and produced by a dry-cast process, then RM was incorporated with 10% tamarind xyloglucan plus gellan gum 1:1 and 10% lysozyme (RMGT-LZ) and with 10% gellan gum and 10% lysozyme (RMG-LZ). Membrane hydrophobic/hydrophilic characteristics were investigated by static/dynamic contact-angle measurements. They were cultivated with MSCs, and cell adhesion, proliferation, and migration capacity was analyzed with MTT assays. Morphological and topographic characteristics were analyzed by scanning electron microscopy. MSC patterns in flow cytometry and differentiation into adipocytes and osteocytes were checked. In vivo assays used RMG-LZ and RMGT-LZ (with and without MSCs) in. In vitro results demonstrated carboxyl and amine groups made the membranes moderately hydrophobic and xyloglucan inclusion decreased wettability, favoring MSC adhesion, proliferation, and differentiation. In vivo, we obtained 40% and 60% reduction in acute/chronic inflammatory infiltrates, 96% decrease in injury area, increased vascular proliferation and collagen deposition, and complete epithelialization after 30 days. MSCs were detected in burned tissue, confirming they had homed and proliferated in vivo.. Nanostructured cellulose-gellan-xyloglucan-lysozyme dressings, especially when seeded with MSCs, improved deep second-degree burn regeneration.

Topics: Animals; Bandages; Blood Vessels; Burns; Cell Adhesion; Cell Differentiation; Cell Proliferation; Cellulose; Collagen; Glucans; Inflammation; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Muramidase; Nanostructures; Polysaccharides, Bacterial; Rats, Wistar; Wound Healing; Xylans

Lysozyme (LZM) is a natural anti-bacterial protein that is found in the saliva, tears and milk of all mammals including humans. Its anti-bacterial properties result from the ability to cleave bacterial cell walls, causing bacterial death. The current study was conducted to investigate the effects of dietary LZM on fecal microbial composition and variation in metabolites in sow. The addition of LZM decreased the fecal short-chain fatty acids (SCFAs). Zonulin and endotoxin in the serum, and feces, were decreased with lysozyme supplementation. Furthermore, fecal concentrations of lipocalin-2 and the pro-inflammatory cytokine TNF-α were also decreased while the anti-inflammatory cytokine IL-10 was increased by lysozyme supplementation. 16S rRNA gene sequencing of the V3-V4 region suggested that fecal microbial levels changed at different taxonomic levels with the addition of LZM. Representative changes included the reduction of diversity between sows, decreased Bacteroidetes, Actinobacteria, Tenericutes and Spirochaetes during lactation as well as an increase in Lactobacillus. These findings suggest that dietary lysozyme supplementation from late gestation to lactation promote microbial changes, which would potentially be the mechanisms by which maternal metabolites and inflammatory status was altered after LZM supplementation.

Topics: Actinobacteria; Animal Feed; Animals; Bacteria; Bacteroidetes; Diet; Dietary Supplements; Fatty Acids, Volatile; Feces; Female; Inflammation; Interleukin-10; Lactation; Lipocalin-2; Muramidase; RNA, Ribosomal, 16S; Spirochaetales; Swine; Tenericutes; Time Factors; Tumor Necrosis Factor-alpha

Supplying immunostimulants to aquatic feed has been an effective way to enhance the health of aquatic animals and substitute for antibiotics. In the present study, the potential effects of Astragalus polysaccharides (APS) were evaluated in turbot, Scophthalmus maximus. Two levels of APS (50 and 150 mg/kg) were added to the basal diet (CON) and a 63-day growth trial (initial weight 10.13 ± 0.04 g) was conducted. As the results showed, significant improvement on growth performance in the APS groups were observed. In addition, dietary 150 mg/kg APS significantly increased the total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-PX) and lysozyme activities in liver. Meanwhile, APS diets induced the mRNA expression of toll-like receptors (TLRs) such as tlr5α, tlr5β, tlr8 and tlr21, while reduced the expression of tlr3 and tlr22. The expression of inflammatory genes myeloid differentiation factor 88 and nuclear factor kappa b p65 and pro-inflammatory cytokines tumor necrosis factor-α and interleukin-1β were up-regulated in APS groups while the expression of anti-inflammatory cytokine transforming growth factor beta was inhibited. Taken together, the present study indicated that Astragalus polysaccharides could remarkably enhance the growth performance, antioxidant activity and maintain an active immune response in turbot.

Topics: Animals; Antioxidants; Astragalus Plant; Body Weight; Dietary Carbohydrates; Dietary Supplements; Flatfishes; Inflammation; Liver; Muramidase; Polysaccharides; Signal Transduction

Although the molecular components of circadian rhythms oscillate in discrete cellular components of the vasculature and many aspects of vascular function display diurnal variation, the cellular connections between the molecular clock and inflammatory cardiovascular diseases remain to be elucidated. Previously we have shown that pre- versus postnatal deletion of Bmal1 (brain and muscle aryl hydrocarbon receptor nuclear translocator-like 1), the nonredundant core clock gene has contrasting effects on atherogenesis. Here we investigated the effect of myeloid cell Bmal1 deletion on atherogenesis and abdominal aortic aneurysm formation in mice. Approach and Results: Mice lacking Bmal1 in myeloid cells were generated by crossing Bmal1 flox/flox mice with lysozyme 2 promoter-driven Cre recombinase mice on a hyperlipidemic low-density lipoprotein receptor-deficient background and were fed on a high-fat diet to induce atherosclerosis. Atherogenesis was restrained, concomitant with a reduction of aortic proinflammatory gene expression in myeloid cell Bmal1 knockout mice. Body weight, blood pressure, blood glucose, triglycerides, and cholesterol were unaltered. Similarly, myeloid cell depletion of Bmal1 also restrained Ang II (angiotensin II) induced formation of abdominal aortic aneurysm in hyperlipidemic mice. In vitro, RNA-Seq analysis demonstrated a proinflammatory response in cultured macrophages in which there was overexpression of Bmal1.. Myeloid cell Bmal1 deletion retards atherogenesis and restrains the formation of abdominal aortic aneurysm and may represent a potential therapeutic target for inflammatory cardiovascular diseases.

Topics: Angiotensin II; Animals; Aortic Aneurysm, Abdominal; ARNTL Transcription Factors; Atherosclerosis; Cells, Cultured; Crosses, Genetic; Diet, High-Fat; Gene Deletion; Gene Expression; Hyperlipidemias; Inflammation; Integrases; Macrophages, Peritoneal; Mice; Mice, Knockout; Muramidase; Myeloid Cells; Promoter Regions, Genetic; Receptors, LDL

Lysozyme is one of the most important anti-bacterial effectors in the innate immune system of animals. Besides its direct antibacterial enzymatic activity, lysozyme displays other biological properties, pointing toward a significant anti-inflammatory effect, many aspects of which are still elusive. Here we investigate the perturbation of gene expression profiles induced by lysozyme in a monocyte cell line in vitro considering a perspective as broad as the whole transcriptome profiling. The results of the RNA-seq experiment show that lysozyme induces transcriptional modulation of the TNF-α/IL-1β pathway genes in U937 monocytes. The analysis of transcriptomic profiles with IPA

Topics: Cell Line; Humans; Inflammation; Interleukin-1beta; Monocytes; Muramidase; Signal Transduction; Transcriptional Activation; Transcriptome; Tumor Necrosis Factor-alpha

Lysozyme is an important component of the innate immune system and has roles in peptidoglycan cleavage of gram-positive organisms. Myeloid cells highly express the isoform, lysozyme M, and its promoter has been used to direct Cre recombinase expression to target deletion of floxed genes in myeloid cells. However, generation of the LysMCre mouse effectively disrupts the LysM gene, and mice homozygous for the Cre allele lack the LysM gene product. To test the contribution of LysM in sterile acute lung injury, we generated LysMCre mice homozygous for the Cre allele (+/+) or wild-type allele (-/-). These mice were challenged with LPS delivered via oropharygneal aspiration. Mice were monitored and weighed daily, and BAL cell counts, differential, protein, and cytokine levels were assessed at days 2 and 4. LysMCre+/+ and LysMCre-/- had similar weight loss and recovery, and similar inflammatory responses to LPS at days 2 and 4. These findings indicate that loss of LysM and expression of Cre recombinase are non-contributory in sterile acute lung injury.

Topics: Acute Lung Injury; Animals; Bronchoalveolar Lavage Fluid; Chemokine CCL2; Chemokine CXCL1; Cytokines; Gene Targeting; Homozygote; Inflammation; Integrases; Lipopolysaccharides; Mice; Muramidase; Myeloid Cells; Real-Time Polymerase Chain Reaction; Recovery of Function; Weight Loss

Topics: Animal Feed; Animals; Bass; Blood Bactericidal Activity; Blood Cell Count; Body Weight; Complement Pathway, Alternative; Disease Resistance; Dose-Response Relationship, Drug; Erythrocyte Indices; Fish Diseases; Gene Expression Profiling; Gram-Negative Bacterial Infections; Hemoglobins; Hydrocortisone; Immunity, Humoral; Inflammation; Muramidase; Neuroimmunomodulation; Nutritional Requirements; Peroxidases; Photobacterium; Tryptophan

Immune homeostasis in the gut associated lymphoid tissues (GALT) is critical to prevent the development of inadvertent pathologies. B cells as the producers of antibodies and cytokines plays an important role in maintaining the GALT homeostasis. However, the mechanism by which B cells specifically direct their responses towards non-self-antigens and become ignorant to self-antigens in the GALT is not known. Therefore, we developed a novel mouse model by expressing Duck Egg Lysozyme (DEL) in gut epithelial cells in presence of HEL reactive B cells. Notably, we observed a transient activation and rapid deletion of self-reactive B cells in Peyers Patches and Mesenteric lymph nodes upon self-antigen exposure. The survival of self-reactive B cells upon exposure to their self-antigen was partially rescued by blocking receptor editing but could be completely rescued by stronger survival signal like ectopic expression of BCL2. Importantly, rescuing the self-reactive B cells promoted production of auto-antibodies and gut inflammation. Mechanistically, we identify a specific activation of TGFβ signaling in self-reactive B cells in the gut and a critical role of this pathway in maintaining peripheral tolerance. Collectively, our studies describe functional consequences and fate of self-reactive B cells in GALT and provide novel mechanistic insights governing self-tolerance of B cells in the gut.

Topics: Animals; Autoantigens; B-Lymphocytes; Bone Marrow; Epithelial Cells; Gastrointestinal Tract; Homeostasis; Inflammation; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Knockout; Models, Animal; Muramidase; Proto-Oncogene Proteins c-bcl-2; Transforming Growth Factor beta

The potent immunomodulatory effect of the endocrine disruptor bisphenol A during development and consequences during life span are of increasing concern. Particular interests have been raised from animal studies regarding the risk of developing food intolerance and infection. We aimed to identify immune disorders in mice triggered by perinatal exposure to bisphenol A. Gravid mice were orally exposed to bisphenol (50 μg/kg body weight/day) from day 15 of pregnancy until weaning. Gut barrier function, local and systemic immunity were assessed in adult female offspring. Mice perinatally exposed to bisphenol showed a decrease in ileal lysozyme expression and a fall of fecal antimicrobial activity. In offspring mice exposed to bisphenol, an increase in colonic permeability was observed associated with an increase in interferon-γ level and a drop of colonic IgA

Topics: Aldehyde Dehydrogenase 1 Family; Animals; Benzhydryl Compounds; Dendritic Cells; Endocrine Disruptors; Feces; Female; Gastrointestinal Tract; Immunity, Humoral; Inflammation; Isoenzymes; Male; Mice, Inbred C3H; Muramidase; Phenols; Pregnancy; Prenatal Exposure Delayed Effects; Retinal Dehydrogenase; Spleen; T-Lymphocytes; Th17 Cells

Feeding small European sea bass, Dicentrarchus labrax, for 6 weeks with Tenebrio molitor larval meal showed significant anti-inflammatory responses (ceruloplasmin, myeloperoxidase and nitric oxide). Serum bacteriolytic activity against a Gram negative bacterium was not significantly affected by dietary Tenebrio, while both lysozyme antibacterial activity and serum trypsin inhibition usually linked to the anti-parasite activity of the fish, were significantly enhanced. The latter may be due to the similarities in the composition of the exoskeleton of parasites and insects that may therefore act as an immunostimulant potentially increasing the anti-parasitic activity. The addition of exogenous proteases significantly decreased both trypsin-inhibition and serum bacteriolytic activity probably through direct inhibition of the proteins responsible for these immune functions. Further investigation involving bacterial or parasitic challenges will be necessary to assess if the effects of dietary mealworm meal on the immune system observed in the present study are translated into an improved resistance to diseases.

Topics: Animals; Anti-Inflammatory Agents; Bass; Ceruloplasmin; Diet; Fish Proteins; Immune System; Immunity, Innate; Inflammation; Insecta; Muramidase; Nitric Oxide; Peroxidase; Tenebrio; Trypsin

Rheumatoid arthritis (RA), a disease that causes joint destruction and bone erosion, is related to osteoclast activity. RA is generally treated with methotrexate (MTX). In this study, a MTX-Alendronate (ALN) conjugate was synthesized and characterized. The conjugate dramatically inhibited osteoclast formation and bone resorption compared with MTX and ALN used alone or in combination. Due to the characteristics of ALN, the MTX-ALN conjugate can adhere to the exposed bone surface and enhance drug accumulation in the pathological region for targeted therapy against osteoclastogenesis. Additionally, MTX was rapidly released in the presence of lysozyme under mildly acidic conditions, similar to inflammatory tissue and osteoclast-surviving conditions, which contributes to inflammatory inhibition; this was confirmed by the presence of pro-inflammatory cytokines. Our study highlights the use of the MTX-ALN conjugate as a potential therapeutic approach for RA by targeting osteoclastogenesis.

Topics: Alendronate; Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Bone and Bones; Bone Resorption; Collagen; Cytokines; Drug Therapy, Combination; Female; Inflammation; Male; Methotrexate; Mice; Mice, Inbred C57BL; Muramidase; Osteoclasts; Osteogenesis; Rats; Rats, Wistar

Aberrant activation of innate immune receptors can cause a spectrum of immune disorders, such as Aicardi-Goutières syndrome (AGS). One such receptor is MDA5, a viral dsRNA sensor that induces antiviral immune response. Using a newly developed RNase-protection/RNA-seq approach, we demonstrate here that constitutive activation of MDA5 in AGS results from the loss of tolerance to cellular dsRNAs formed by Alu retroelements. While wild-type MDA5 cannot efficiently recognize Alu-dsRNAs because of its limited filament formation on imperfect duplexes, AGS variants of MDA5 display reduced sensitivity to duplex structural irregularities, assembling signaling-competent filaments on Alu-dsRNAs. Moreover, we identified an unexpected role of an RNA-rich cellular environment in suppressing aberrant MDA5 oligomerization, highlighting context dependence of self versus non-self discrimination. Overall, our work demonstrates that the increased efficiency of MDA5 in recognizing dsRNA comes at a cost of self-recognition and implicates a unique role of Alu-dsRNAs as virus-like elements that shape the primate immune system.

Topics: A549 Cells; Alu Elements; Autoimmune Diseases of the Nervous System; Humans; Inflammation; Interferon-Induced Helicase, IFIH1; Muramidase; Nervous System Malformations; Peptide Fragments; Protein Multimerization; RNA, Double-Stranded; Self Tolerance; THP-1 Cells

Obesity is an established risk factor for many diseases including intestinal cancer. One of the responsible mechanisms is the chronic inflammation driven by obesity. However, it remains to be defined whether diet-induced obesity exacerbates the intestinal inflammatory status by cytokines produced in adipose tissue or the high fat diet first alters the gut microbiota and then drives intestinal inflammation. To address this question, we fed C57BL/6 mice with a high fat diet (HF, 60%) and sacrificed them sequentially after 8, 12, and 16 weeks, and then compositions of gut microbiota and expressions of antimicrobial peptides were determined. The compositions of gut microbiota were altered at 8 wk HF feeding, followed with reduced Paneth antimicrobial peptides lysozyme and Reg III

Topics: Animals; Blotting, Western; Cytokines; Diet, High-Fat; Gastrointestinal Microbiome; Immunohistochemistry; Inflammation; Male; Mice; Mice, Inbred C57BL; Muramidase; Obesity; Paneth Cells; Peptides; Ribonuclease, Pancreatic

The goal of this investigation is to analyze the effect of therapeutic low-level laser therapy (LLLT) to have a possibility to check pain and inflammation connected with surgical removal of impacted lower third molars in general anesthesia or even phobia [not accompanied by pain or fear of dental treatment, using immunologic markers-secretory immunoglobulin A (sIgA) and lysozyme]. The healing process was also monitored by infrared thermography.. LLLT can accelerate the proliferation phase of healing and decrease the inflammatory reaction, but the effect is not really clear.. The treatment group comprised 213 impacted third molars (144 laser group and 74 placebo group). Laser radiation (diode laser 830 nm) was applied. The effectivity of laser therapy was evaluated based on immunological tests, that is, before and after treatment with sIgA and lysozyme in nonstimulated saliva. Thermographic examination was performed by infrared camera.. After laser irradiation, the sIgA decreases from 546.91 mg/L (SD 354.58) to 304.91 mg/L (SD 191.96), and in the control group from 602.25 mg/L (SD 343.62) to 425.62 mg/L (SD 220.51); the differences were significant, the lysozyme value being lower. After laser therapy, the laser and placebo sides in the area of the third molars were differed in 0.2°C.. The 830 nm wavelength penetrates to deep-seated tissues. A positive association was found between concentration of salivary sIgA and lysozyme in the saliva after LLLT application. The deep-seated wounds after wisdom teeth extraction had no effect on temperature rise in the face.

Topics: Adolescent; Adult; Female; Humans; Immunoglobulin A; Inflammation; Low-Level Light Therapy; Male; Molar, Third; Muramidase; Pain Measurement; Pain, Postoperative; Saliva; Thermography; Tooth Extraction; Tooth, Impacted; Treatment Outcome; Wound Healing

Ocular antigens are sequestered behind the blood-retina barrier and the ocular environment protects ocular tissues from autoimmune attack. The signals required to activate autoreactive T cells and allow them to cause disease in the eye remain in part unclear. In particular, the consequences of peripheral presentation of ocular antigens are not fully understood. We examined peripheral expression and presentation of ocular neo-self-antigen in transgenic mice expressing hen egg lysozyme (HEL) under a retina-specific promoter. High levels of HEL were expressed in the eye compared to low expression throughout the lymphoid system. Adoptively transferred naïve HEL-specific CD4

Topics: Animals; Antigen Presentation; Autoantigens; Autoimmunity; CD4-Positive T-Lymphocytes; Cell Proliferation; Cross Reactions; Inflammation; Mice; Muramidase; Retina; Risk

Topics: Diclofenac; Electrodes; Humans; Inflammation; Molecular Docking Simulation; Muramidase; Thermodynamics

Acne vulgaris is the most common skin disorder, and is caused by Propionibacterium acnes (P. acnes) and can induce inflammation. Antibiotic therapy often needs to be administered for long durations in acne therapy, which results in extensive antibiotic exposure. The present study investigated a new treatment model for evaluating the antibacterial effects of lysozyme (LY)-shelled microbubbles (MBs) and ultrasound (US)-mediated LY-shelled MBs cavitation against P. acnes both in vitro and in vivo, with the aims of reducing the dose and treatment duration and improving the prognosis of acne vulgaris. In terms of the in vitro treatment efficacy, the growth of P. acnes was inhibited by 86.08 ± 2.99% in the LY-shelled MBs group and by 57.74 ± 3.09% in the LY solution group. For US power densities of 1, 2, and 3 W/cm

Topics: Animals; Anti-Bacterial Agents; Chickens; Colony Count, Microbial; Inflammation; Mice, Inbred ICR; Microbial Sensitivity Tests; Microbubbles; Muramidase; Propionibacterium acnes; Skin Diseases; Ultrasonography

Polymorphisms in the PTPN11 gene encoding for the tyrosine phosphatase SHP-2 were described in patients with ulcerative colitis. We have recently demonstrated that mice with an intestinal epithelial cell-specific deletion of SHP-2 (SHP-2(IEC-KO) ) develop severe colitis 1 month after birth. However, the mechanisms by which SHP-2 deletion induces colonic inflammation remain to be elucidated. We generated SHP-2(IEC-KO) mice lacking Myd88 exclusively in the intestinal epithelium. The colonic phenotype was histologically analyzed and cell differentiation was determined by electron microscopy and lysozyme or Alcian blue staining. Microbiota composition was analyzed by 16S sequencing. Results show that innate defense genes including those specific to Paneth cells were strongly up-regulated in SHP-2-deficient colons. Expansion of intermediate cells (common progenitors of the Goblet and Paneth cell lineages) was found in the colon of SHP-2(IEC-KO) mice whereas Goblet cell number was clearly diminished. These alterations in Goblet/intermediate cell ratio were noticed 2 weeks after birth, before the onset of inflammation and were associated with significant alterations in microbiota composition. Indeed, an increase in Enterobacteriaceae and a decrease in Firmicutes were observed in the colon of these mice, indicating that dysbiosis also occurred prior to inflammation. Importantly, loss of epithelial Myd88 expression inhibited colitis development in SHP-2(IEC-KO) mice, rescued Goblet/intermediate cell ratio, and prevented NFκB hyperactivation and inflammation. These data indicate that SHP-2 is functionally important for the maintenance of appropriate barrier function and host-microbiota homeostasis in the large intestine. J. Cell. Physiol. 231: 2529-2540, 2016. © 2016 The Authors. Journal of Cellular Physiology published by Wiley Periodicals, Inc.

Topics: Animals; Animals, Newborn; Antimicrobial Cationic Peptides; Biomarkers; Cell Differentiation; Colon; Goblet Cells; Homeostasis; Inflammation; Mice, Inbred C57BL; Microbiota; Muramidase; Myeloid Differentiation Factor 88; Paneth Cells; Protein Tyrosine Phosphatase, Non-Receptor Type 11; Up-Regulation

Resident microglia and infiltrating myeloid cells play important roles in the onset, propagation, and resolution of inflammation in central nervous system (CNS) injury and disease. Identifying cell type-specific mechanisms will help to appropriately target interventions for tissue repair. Arginase-1 (Arg-1) is a well characterised modulator of tissue repair and its expression correlates with recovery after CNS injury. Here we assessed the cellular localisation of Arg-1 in two models of CNS damage. Using microglia specific antibodies, P2ry12 and Fc receptor-like S (FCRLS), we show the LysM-EGFP reporter mouse is an excellent model to distinguish infiltrating myeloid cells from resident microglia. We show that Arg-1 is expressed exclusively in infiltrating myeloid cells but not microglia in models of spinal cord injury (SCI) and experimental autoimmune encephalomyelitis (EAE). Our in vitro studies suggest that factors in the CNS environment prevent expression of Arg-1 in microglia in vivo. This work suggests different functional roles for these cells in CNS injury and repair and shows that such repair pathways can be switched on in infiltrating myeloid cells in pro-inflammatory environments.

Topics: Animals; Arginase; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Green Fluorescent Proteins; Inflammation; Mice; Mice, Inbred C57BL; Microglia; Muramidase; Myeloid Cells; Spinal Cord Injuries

Interleukin (IL)-8 is the key agent for initiating an inflammatory response to infection with Helicobacter pylori. Some strains of Lactobacillus spp. are known to colonize the stomach and suppress inflammation caused by H. pylori. In this study, we characterized two gastric-derived lactobacilli, Lactobacillus salivarius (LS) strains B37 and B60, capable of inhibiting H. pylori-induced IL-8 production by gastric epithelial cells.. Conditioned media from LS-B37 and LS-B60 suppressed H. pylori-induced IL-8 production and mRNA expression from AGS cells without inhibiting H. pylori growth. These conditioned media suppressed the activation of NF-κB but did not suppress c-Jun activation. IL-8 inhibitory substances in conditioned media of LS-B37 and LS-B60 are heat-stable and larger than 100 kDa in size. The inhibitory activity of LS-B37 was abolished when the conditioned medium was treated with α-amylase but still remained when treated with either proteinase K, trypsin, lipase or lysozyme. The activity of LS-B60 was abolished when the conditioned medium was treated with either amylase or proteinase K but still remained when treated with lysozyme. Treatment with lipase and trypsin also significantly affected the inhibitory activity of LS-B60 although the conditioned medium retained IL-8 suppression statistically different from media control.. These results suggest that L. salivarius strains B37 and B60 produce different immunomodulatory factors capable of suppressing H. pylori-induced IL-8 production from gastric epithelial cells. Our results suggest that the large, heat-stable immunomodulatory substance(s) present in the LCM of LS-B37 is a polysaccharide, while the one(s) of LS-B60 is either complex consisting of components of polysaccharide, lipid and protein or includes multiple components such as glycoprotein and lipoprotein.

Topics: alpha-Amylases; Anti-Inflammatory Agents; Cell Line; Culture Media, Conditioned; Endopeptidase K; Epithelial Cells; Gastric Mucosa; Gene Expression Regulation, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; Lactobacillus; Ligilactobacillus salivarius; Lipase; Muramidase; NF-kappa B; Probiotics; RNA, Messenger; Stomach; Trypsin

We hypothesized that if internalization of Staphylococcus aureus could be blocked by using cytochalasin D (an inhibitor of phagocytosis and phagolysosome fusion), then the intracellular entry and survival of the pathogen in host's phagocytic cells recruited to the inflammatory site can be restricted. At the same time, if we use antimicrobial agents (e.g., ciprofloxacin and azithromycin) having potent intracellular and extracellular microbicidal activity against the bacterium that have not entered into the phagosome and remains adhered to the phagocytic cell membrane, then they can be eradicated from the site of infection without compromising the host cell. To validate this, role of ciprofloxacin (CIP) and azithromycin (AZM) in eliminating S. aureus by suppressing the phagocytic activity of macrophages with cytochalasin D before infection was investigated. CIP and AZM were used either alone or in combination with cytochalasin D. Supernatant and lysate obtained from the culture of macrophages were used for quantification of reactive oxygen species, lysozymes, antioxidant enzymes, and cytokines produced. Azithromycin was better than ciprofloxacin in combination with cytochalasin D for eradicating S. aureus and regulating cytokine release. Further studies are required for ensuring proper delivery of this combination at the site of infection.

Topics: Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents; Antioxidants; Azithromycin; Ciprofloxacin; Cytochalasin D; Drug Therapy, Combination; Glutathione; Hydrogen Peroxide; Inflammation; Interferon-gamma; Interleukin-10; Interleukin-6; Macrophages; Male; Mice; Microbial Sensitivity Tests; Muramidase; Phagocytosis; Staphylococcal Infections; Staphylococcus aureus; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Chronic low-grade inflammation of adipose tissue plays a crucial role in the pathophysiology of obesity. Immunohistological microscopic analysis in obese fat tissue has demonstrated the infiltration of several immune cells such as macrophages, but dynamics of immune cells have not been fully elucidated and clarified. Here, by using intravital multiphoton imaging technique, to our knowledge for the first time, we analyzed and visualized the inflammatory processes in adipose tissue under high-fat and high-sucrose (HF/HS) diet with lysozyme M-EGFP transgenic (LysM(EGFP)) mice whose EGFP was specifically expressed in the myelomonocytic lineage. Mobility of LysM(EGFP)-positive macrophages was shown to be activated just 5 d after HF/HS diet, when the distinct hypertrophy of adipocytes and the accumulation of macrophages still have not become prominent. Significant increase of S100A8 was detected in mature adipocyte fraction just 5 d after HF/HS diet. Recombinant S100A8 protein stimulated chemotactic migration in vitro and in vivo, as well as induced proinflammatory molecules, both macrophages and adipocytes, such as TNF-α and chemokine (C-C motif) ligand 2. Finally, an antibody against S100A8 efficiently suppressed the HF/HS diet-induced initial inflammatory change, i.e., increased mobilization of adipose LysM(EGFP)-positive macrophages, and ameliorated HF/HS diet-induced insulin resistance. In conclusion, time-lapse intravital multiphoton imaging of adipose tissues identified the very early event exhibiting increased mobility of macrophages, which may be triggered by increased expression of adipose S100A8 and results in progression of chronic inflammation in situ.

Topics: 3T3-L1 Cells; Adipocytes; Adiposity; Animals; Antibodies; Calgranulin A; Chemotaxis; Diet, High-Fat; Epididymis; Green Fluorescent Proteins; Inflammation; Insulin; Lipopolysaccharides; Macrophages; Male; Mice; Microscopy, Fluorescence, Multiphoton; Muramidase; Obesity; RNA, Messenger; Up-Regulation

Intestinal mucosal immune components and mRNA levels of inflammatory cytokines, tight junction proteins, antioxidant enzymes and related signalling molecules in young grass carp (Ctenopharyngodon idellus) under dietary manganese (Mn) deficiency or excess were investigated. Fish were fed the diets containing graded levels of Mn [3.65-27.86 mg Mn kg(-1) diet] for 8 weeks. The results demonstrated that Mn deficiency significantly decreased the lysozyme and acid phosphatase (ACP) activities, up-regulated tumour necrosis factor α (TNF-α), interleukin 8 and the signalling factor nuclear factor-κB p65, and down-regulated interleukin 10 (IL-10), transforming growth factor β1, inhibitor of signalling factors κB-α and target of rapamycin mRNA levels in the proximal intestine (PI), mid intestine (MI) and distal intestine (DI). However, Mn deficiency did not change the C3 content in the PI, whereas it decreased the C3 contents in the MI and DI. Additionally, Mn depletion also resulted in significantly low mRNA levels for tight junction proteins (claudin-b, claudin-c, claudin-15, occludin and zonula occludens-1), antioxidant enzymes (MnSOD, GPx and CAT) and NF-E2-related factor-2 in the intestines of fish. Excessive Mn exhibited toxic effects similar to Mn deficiency, where optimal Mn contents reversed those indicators. In conclusion, Mn deficiency or excess causes the depression of intestinal immunity, induction of inflammation and dysfunction of the intestinal physical barrier relating to NF-κB, TOR and Nrf2 signalling in grass carp. Furthermore, quadratic regression analysis at 95% maximum response of lysozyme and acid phosphatase activities in the distal intestine of young grass carp revealed the optimum dietary Mn levels to be 8.90 and 8.99 mg kg(-1) diet, respectively.

Topics: Acid Phosphatase; Animals; Carps; Complement C3; Cytokines; Diet; Fish Proteins; Inflammation; Intestinal Mucosa; Manganese; Muramidase; NF-E2-Related Factor 2; NF-kappa B; RNA, Messenger; Signal Transduction; Tight Junction Proteins; TOR Serine-Threonine Kinases

Increasing evidence involves sustained pro-inflammatory microglia activation in the pathogenesis of different neurodegenerative diseases, particularly Parkinson's disease (PD). We recently uncovered a completely novel and unexpected role for caspase-8 and its downstream substrates caspase-3/7 in the control of microglia activation and associated neurotoxicity to dopaminergic cells. To demonstrate the genetic evidence, mice bearing a floxed allele ofCASP8 were crossed onto a transgenic line expressing Cre under the control of Lysozyme 2 gene. Analysis of caspase-8 gene deletion in brain microglia demonstrated a high efficiency in activated but not in resident microglia. Mice were challenged with lipopolysaccharide, a potent inducer of microglia activation, or with MPTP, which promotes specific dopaminergic cell damage and consequent reactive microgliosis. In neither of these models, CASP8 deletion appeared to affect the overall number of microglia expressing the pan specific microglia marker, Iba1. In contrast, CD16/CD32 expression, a microglial pro-inflammatory marker, was found to be negatively affected upon CASP8 deletion. Expression of additional proinflammatory markers were also found to be reduced in response to lipopolysaccharide. Of importance, reduced pro-inflammatory microglia activation was accompanied by a significant protection of the nigro-striatal dopaminergic system in the MPTP mouse model of PD.

Topics: Animals; Calcium-Binding Proteins; Caspase 8; Dopaminergic Neurons; Gliosis; Inflammation; Lipopolysaccharides; Mice; Mice, Knockout; Mice, Transgenic; Microfilament Proteins; Microglia; MPTP Poisoning; Muramidase; Myeloid Cells

Although caspase-8 is a well-established initiator of apoptosis and suppressor of necroptosis, recent evidence suggests that this enzyme maintains functions beyond its role in cell death. As cells of the innate immune system, and in particular macrophages, are now at the forefront of autoimmune disease pathogenesis, we examined the potential involvement of caspase-8 within this population.. Cre (LysM) Casp8 (fl/fl) mice were bred via a cross between Casp8 (fl/fl) mice and Cre (LysM) mice, and RIPK3 (-/-) Cre (LysM) Casp8 (fl/fl) mice were generated to assess the contribution of receptor-interacting serine-threonine kinase (RIPK)3. Immunohistochemical and immunofluorescence analyses were used to examine renal damage. Flow cytometric analysis was employed to characterize splenocyte distribution and activation. Cre (LysM) Casp8 (fl/fl) mice were treated with either Toll-like receptor (TLR) agonists or oral antibiotics to assess their response to TLR activation or TLR agonist removal. Luminex-based assays and enzyme-linked immunosorbent assays were used to measure cytokine/chemokine and immunoglobulin levels in serum and cytokine levels in cell culture studies. In vitro cell culture was used to assess macrophage response to cell death stimuli, TLR activation, and M1/M2 polarization. Data were compared using the Mann-Whitney U test.. Loss of caspase-8 expression in macrophages promotes onset of a mild systemic inflammatory disease, which is preventable by the deletion of RIPK3. In vitro cell culture studies reveal that caspase-8-deficient macrophages are prone to a caspase-independent death in response to death receptor ligation; yet, caspase-8-deficient macrophages are not predisposed to unchecked survival, as analysis of mixed bone marrow chimeric mice demonstrates that caspase-8 deficiency does not confer preferential expansion of myeloid populations. Loss of caspase-8 in macrophages dictates the response to TLR activation, as injection of TLR ligands upregulates expression of costimulatory CD86 on the Ly6C(high)CD11b(+)F4/80(+) splenic cells, and oral antibiotic treatment to remove microbiota prevents splenomegaly and lymphadenopathy in Cre (LysM) Casp8 (fl/fl) mice. Further, caspase-8-deficient macrophages are hyperresponsive to TLR activation and exhibit aberrant M1 macrophage polarization due to RIPK activity.. These data demonstrate that caspase-8 functions uniquely in macrophages by controlling the response to TLR activation and macrophage polarization in an RIPK-dependent manner.

Topics: Animals; Antigens, Differentiation; Antigens, Ly; B7-2 Antigen; Blotting, Western; Caspase 8; CD11b Antigen; Cells, Cultured; Female; Flow Cytometry; Inflammation; Macrophage Activation; Macrophages; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Muramidase; Myeloid Cells; Receptor-Interacting Protein Serine-Threonine Kinases; Spleen; Toll-Like Receptors

In our model system using the THP-1 monocytic cell line, whole heat-killed cells of Alloiococcus otitidis elicited several pro-inflammatory cytokines identified in ear effusions of children with otitis media (OM). Levels of these cytokines were equivalent to or greater than those elicited by a standard Gram-positive otopathogen, Streptococcus pneumoniae. The current study examined the hypothesis that extracellular material produced by A. otitidis might also contribute to the inflammatory responses in OM. Cell-free culture filtrates of recent A. otitidis isolates (n = 39) were tested for induction of pro-inflammatory cytokines from THP-1 cells primed with IFN-γ. The highest responses were from IL-8 followed by IL-1β, and the lowest from IL-6. Filtrates from nine isolates were treated with lysozyme or proteinase K to assess the nature of the extracellular stimulants. Peptidoglycan was not a major component eliciting the responses. There was no correlation between colony type or β-haemolysin production. Proteinase K treatment indicated extracellular proteins might induce the inflammatory responses, particularly the 70-75 ku band. Further studies on the role of the extracellular proteins of A. otitidis and cytokine responses in pathogenesis of ear infections are needed.

Topics: Bacterial Proteins; Carnobacteriaceae; Cell Line; Cytokines; Endopeptidase K; Hemolysin Proteins; Humans; Hydrolysis; Inflammation; Interferon-gamma; Monocytes; Muramidase; Otitis Media

Neutrophil lifespan and function are regulated by hypoxia via components of the hypoxia inducible factor (HIF)/von Hippel Lindau/hydroxylase pathway, including specific roles for HIF-1α and prolyl hydroxylase-3. HIF-2α has both distinct and overlapping biological roles with HIF-1α and has not previously been studied in the context of neutrophil biology. We investigated the role of HIF-2α in regulating key neutrophil functions. Human and murine peripheral blood neutrophils expressed HIF-2α, with expression up-regulated by acute and chronic inflammatory stimuli and in disease-associated inflammatory neutrophil. HIF2A gain-of-function mutations resulted in a reduction in neutrophil apoptosis both ex vivo, through the study of patient cells, and in vivo in a zebrafish tail injury model. In contrast, HIF-2α-deficient murine inflammatory neutrophils displayed increased sensitivity to nitrosative stress induced apoptosis ex vivo and increased neutrophil apoptosis in vivo, resulting in a reduction in neutrophilic inflammation and reduced tissue injury. Expression of HIF-2α was temporally dissociated from HIF-1α in vivo and predominated in the resolution phase of inflammation. These data support a critical and selective role for HIF-2α in persistence of neutrophilic inflammation and provide a platform to dissect the therapeutic utility of targeting HIF-2α in chronic inflammatory diseases.

Topics: Animals; Apoptosis; Basic Helix-Loop-Helix Transcription Factors; Cell Hypoxia; Gene Expression Regulation; Green Fluorescent Proteins; Humans; Immunohistochemistry; Inflammation; Mice; Mice, Inbred C57BL; Muramidase; Neutrophils; Phagocytosis; Phenotype; Respiratory Burst; RNA; Zebrafish

Early diagnosis of neurological disorders would greatly improve their management and treatment. A major hurdle is that inflammatory products of cerebral disease are not easily detected in blood. Inflammation in multiple organs and heterogeneity in disease present additional challenges in distinguishing the extent to which a blood-based marker reflects disease in brain or other afflicted organs. Murine models of the monogenetic disorder Niemann-Pick Type C present aggressive forms of cerebral and liver inflammatory disease. Microarray analyses previously revealed age-dependent changes in innate immunity transcripts in the mouse brain. We have now validated four putative secretory inflammatory markers that are also elevated in mouse liver. We include limited, first time analysis of human Niemann-Pick Type C liver and cerebellum. Furthermore, we utilized 2-hydroxypropyl-β-cyclodextrin (HPβCD, an emerging therapeutic) administered intraperitoneally in mice, which abrogates inflammatory pathology in the liver but has limited effect on the brain. By analyzing the corresponding effects on inflammatory plasma proteins, we identified cathepsin S as a lead indicator of liver disease. In contrast, lysozyme was a marker of both brain and liver disease. 2-Hydroxypropyl-β-cyclodextrin had no effect on transcripts of neuron-specific 24-hydroxylase, and its product 24(S)-hydroxycholesterol was not a useful indicator in mouse plasma. Our data suggest that dual analysis of levels of the inflammatory markers lysozyme and cathepsin S may enable detection of multiple distinct states of neurodegeneration in plasma.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Animals; beta-Cyclodextrins; Brain; Cathepsins; Disease Models, Animal; Female; Gene Deletion; Humans; Inflammation; Intracellular Signaling Peptides and Proteins; Liver; Male; Mice; Mice, Inbred BALB C; Muramidase; Niemann-Pick C1 Protein; Niemann-Pick Disease, Type C; Proteins

Frogs are a diverse group of vertebrates for which limited genomic resources are available. Natural frog populations face a multitude of threats, including habitat degradation, infectious disease, and environmental change. Characterizing the functional genomics of anuran tissues in general - and the immune system in particular - will enhance our knowledge of genetic and epigenetic responses to environmental threats and inform conservation and recovery efforts.. To increase the number of species with genomic datasets and characterize gene expression in immune-related tissues, we sequenced the transcriptomes of three tissues from two frogs (Espadarana prosoblepon and Lithobates yavapaiensis) on the Roche 454 GS FLX platform. Our sequencing produced 8881 E. prosoblepon and 5428 L. yavapaiensis annotated gene products after de novo assembly and Gene Ontology classification. Transcripts of the innate and acquired immune system were expressed in all three tissues. Inflammatory response and acquired immunity transcripts were significantly more diverged between E. prosoblepon and L. yavapaiensis compared to innate immunity and immune system development transcripts. Immune-related transcripts did not show an overall elevated rate of functional evolution, with the exception of glycosyl proteases, which include lysozymes, central bacterial and fungal-killing enzymes of the innate immune system.. The three frog transcriptomes provide more than 600 Mbp of new genomic data, and will serve as a valuable framework for future comparative studies of non-model anurans. Additionally, we show that immune gene divergence varies by functional group and that transcriptome studies can be useful in comparing rates of evolutionary change across gene families.

Topics: Animals; Anura; Costa Rica; Evolution, Molecular; Female; Gene Ontology; Genome; High-Throughput Nucleotide Sequencing; Immune System; Inflammation; Intestines; Male; Muramidase; Mycoses; Panama; Polymorphism, Single Nucleotide; Proteins; Ranidae; Skin; Spleen; Transcriptome

Understanding the interplay between adiposity, inflammation, and cardiovascular complications in type 2 diabetes mellitus (T2DM) remains a challenge. Signaling from adipocytes is considered important in this context. Adiponectin is the most abundant adipocytokine and has been associated with various measures of cardiovascular disease (CVD). This study examines the relationships between genetic variants in the adiponectin (ADIPOQ) and adiponectin-related signaling pathway genes and measures of subclinical CVD (vascular calcified plaque and carotid intima-media thickness), plasma lipids, and inflammation in T2DM.. Single-nucleotide polymorphisms (SNPs) in ADIPOQ (n = 45), SNPs tagging ADIPOR1 (n = 6), APIPOR2 (n = 8), APPL1 (n = 6) and known rare coding variants in KNG1 (n = 3) and LYZL1 (n = 3) were genotyped in 1220 European Americans from the family-based Diabetes Heart Study. Associations between SNPs and phenotypes of interest were assessed using a variance components analysis with adjustment for age, sex, T2DM-affected status, and body mass index.. There was minimal evidence of association between SNPs in the adiponectin signaling pathway genes and measures of calcified plaque; eight of the 71 SNPs showed evidence of association with subclinical CVD (P = 0.007-0.046) but not with other phenotypes examined. Nine additional SNPs were associated with at least one of the plasma lipid measures (P = 0.008-0.05).. Findings from this study do not support a significant role for variants in the adiponectin signaling pathway genes in contributing to risk for vascular calcification in T2DM. However, further understanding the interplay between adiposity, plasma lipids, and inflammation may prove important in the prediction and management of cardiovascular complications in T2DM.

Topics: Adaptor Proteins, Signal Transducing; Adiponectin; Aged; Cardiovascular Diseases; Diabetes Complications; Diabetes Mellitus, Type 2; Female; Genotype; Humans; Inflammation; Lipids; Male; Middle Aged; Muramidase; Obesity; Phenotype; Plaque, Atherosclerotic; Polymorphism, Single Nucleotide; Receptors, Adiponectin; Signal Transduction; Tunica Intima; Tunica Media; Vascular Calcification; White People

The spread of drug-resistant bacterial pathogens is a growing global concern and has prompted an effort to explore potential adjuvant and alternative therapies derived from nature's repertoire of bactericidal proteins and peptides. In humans, the airway surface liquid layer is a rich source of antibiotics, and lysozyme represents one of the most abundant and effective antimicrobial components of airway secretions. Human lysozyme is active against both Gram-positive and Gram-negative bacteria, acting through several mechanisms, including catalytic degradation of cell wall peptidoglycan and subsequent bacterial lysis. In the infected lung, however, lysozyme's dense cationic character can result in sequestration and inhibition by polyanions associated with airway inflammation. As a result, the efficacy of the native enzyme may be compromised in the infected and inflamed lung. To address this limitation, we previously constructed a charge-engineered variant of human lysozyme that was less prone to electrostatic-mediated inhibition in vitro. Here, we employ a murine model to show that this engineered enzyme is superior to wild-type human lysozyme as a treatment for mucoid Pseudomonas aeruginosa lung infections. The engineered enzyme effectively decreases the bacterial burden and reduces markers of inflammation and lung injury. Importantly, we found no evidence of acute toxicity or allergic hypersensitivity upon repeated administration of the engineered biotherapeutic. Thus, the charge-engineered lysozyme represents an interesting therapeutic candidate for P. aeruginosa lung infections.

Topics: Animals; Anti-Bacterial Agents; Bronchoalveolar Lavage Fluid; Colony Count, Microbial; Cytokines; Glycosaminoglycans; Humans; Inflammation; Lung; Male; Mice; Mice, Inbred C57BL; Muramidase; Protein Engineering; Pseudomonas aeruginosa; Pseudomonas Infections; Static Electricity

Sickness behaviors, such as fatigue, mood alterations, and cognitive dysfunction, which result from changes in central neurotransmission, are prevalent in systemic inflammatory diseases and greatly impact patient quality of life. Although, microglia (resident cerebral immune cells) and cytokines (e.g., TNFα) are associated with changes in central neurotransmission, the link between peripheral organ inflammation, circulating cytokine signaling, and microglial activation remains poorly understood. Here we demonstrate, using cerebral intravital microscopy, that in response to liver inflammation, there is increased monocyte specific rolling and adhesion along cerebral endothelial cells (CECs). Peripheral TNFα-TNFR1 signaling and the adhesion molecule P-selectin are central mediators of these monocyte-CEC adhesive interactions which were found to be closely associated with microglial activation, decreased central neural excitability and sickness behavior development. Similar monocyte-CEC adhesive interactions were also observed in another mouse model of peripheral organ inflammation (i.e., 2,4-dinitrobenzene sulfonic acid-induced colitis). Our observations provide a clear link between peripheral organ inflammation and cerebral changes that impact behavior, which can potentially allow for novel therapeutic interventions in patients with systemic inflammatory diseases.

Topics: Alanine Transaminase; Animals; Cell Adhesion; Cerebral Cortex; Cholestasis; Colitis; Cytokines; Dinitrofluorobenzene; Disease Models, Animal; Endothelial Cells; Female; Hippocampus; Illness Behavior; Inflammation; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Monocytes; Muramidase; P-Selectin; Pentylenetetrazole

Depression is accompanied by activation of immuno-inflammatory and oxidative and nitrosative stress (IO&NS) pathways, and increased IgM/IgA responses to lipopolysaccharide (LPS) of gram-negative commensal bacteria. The latter suggests that bacterial translocation has caused IgM/IgA responses directed against LPS. Bacterial translocation may drive IO&NS responses.. To examine the associations between IgM/IgA responses to LPS and IO&NS measurements, including plasma/serum interleukin-1 (IL-1), tumor necrosis factor (TNF)α, neopterin, lysozyme, oxidized LDL (oxLDL) antibodies, peroxides, and IgM (auto)immune responses against malondialdehyde (MDA), azelaic acid, phophatidyl inositol (Pi), NO-tryptophan and NO-tyrosine in depressed patients and controls.. We found significant positive associations between IgM/IgA responses to LPS and oxLDL antibodies, IgM responses against MDA, azelaic acid, Pi, NO-tryptophan, and NO-tyrosine. The IgA responses to LPS were correlated with lysozyme. There were no significant positive correlations between the IgM/IgA responses to LPS and IL-1 and neopterin.. The findings show that in depression there is an association between increased bacterial translocation and lysozyme production, an antibacterial compound, O&NS processes, and autoimmune responses directed against O&NS generated neoantigenic determinants. It is suggested that bacterial translocation may drive IO&NS pathways in depression and thus play a role in its pathophysiology.

Topics: Adult; Autoimmunity; Bacterial Translocation; Case-Control Studies; Cross-Sectional Studies; Depressive Disorder, Major; Epitopes; Female; Humans; Immunoglobulin A; Immunoglobulin M; Inflammation; Interleukin-1; Lipopolysaccharides; Male; Muramidase; Neopterin; Nervous System; Oxidative Stress; Tumor Necrosis Factor-alpha

Inflammation occurs in many amyloidoses, but its underlying mechanisms remain enigmatic. Here we show that amyloid fibrils of human lysozyme, which are associated with severe systemic amyloidoses, induce the secretion of pro-inflammatory cytokines through activation of the NLRP3 (NLR, pyrin domain containing 3) inflammasome and the Toll-like receptor 2, two innate immune receptors that may be involved in immune responses associated to amyloidoses. More importantly, our data clearly suggest that the induction of inflammatory responses by amyloid fibrils is linked to their intrinsic structure, because the monomeric form and a non-fibrillar type of lysozyme aggregates are both unable to trigger cytokine secretion. These lysozyme species lack the so-called cross-β structure, a characteristic structural motif common to all amyloid fibrils irrespective of their origin. Since fibrils of other bacterial and endogenous proteins have been shown to trigger immunological responses, our observations suggest that the cross-β structural signature might be recognized as a generic danger signal by the immune system.

Topics: Amyloid; Amyloidosis; Animals; Carrier Proteins; Cell Line; HEK293 Cells; Humans; Immunity, Innate; Inflammasomes; Inflammation; Interleukin-1beta; Mice; Mice, Inbred C57BL; Muramidase; NLR Family, Pyrin Domain-Containing 3 Protein; Protein Structure, Secondary; Toll-Like Receptor 2

Sleep-disordered breathing (SDB) is a risk factor for cardiovascular disease (CVD). The underlying pathogenesis is not clear. In patients with obstructive sleep apnoea syndrome (OSAS) elevated levels of inflammatory markers, such as C-reactive protein (CRP), interleukin-6 (IL-6) and tumour necrosis factor α (TNFα) have been found. These markers have also been shown as independent markers of CVD in other populations. The aim of the study was to investigate the association between SDB and systemic inflammation in a population-based cohort of women. From 6817 women who previously answered a questionnaire concerning snoring habits, 230 habitually snoring women and 170 women regardless of snoring status went through polysomnography, anthropometric measurements and blood sampling. Analyses were made for CRP, TNFα, IL-6, myeloperoxidase (MPO) and lysozyme. The levels of CRP, IL-6 and lysozyme were significantly higher in subjects with apnoea-hypopnoea index (AHI) ≥15 compared with women with lower AHI. All inflammatory markers except MPO correlated to AHI and oxygen desaturation measures, and to waist circumference. In multiple linear regressions adjusting for age, waist circumference and smoking, independent correlations between oxygen desaturation indices (ODI) and inflammation were found for IL-6 (P = 0.03 for % sleep time with saturation <90%) and TNFα (P = 0.03 for ODI 3%). No significant correlations were found between AHI and inflammation. Also, for women from the general population there is an independent correlation between SDB and inflammation, even after adjusting for obesity. The results indicate that intermittent hypoxia, and not the AHI, is related to systemic inflammation seen in OSAS.

Topics: Adult; Aged; Biomarkers; C-Reactive Protein; Female; Humans; Inflammation; Interleukin-6; Middle Aged; Muramidase; Peroxidase; Polysomnography; Sleep Apnea Syndromes; Snoring; Surveys and Questionnaires; Tumor Necrosis Factor-alpha; Young Adult

There is evidence that inflammatory pathways and cell-mediated immunity (CMI) play an important role in the pathophysiology of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS). Activation of inflammatory and CMI pathways, including increased levels of cytokines, is known to induce fatigue and somatic symptoms. Given the broad spectrum inflammatory state in ME/CFS, the aim of this study was to examine whether inflammatory and CMI biomarkers are increased in individuals with ME/CFS.. In this study we therefore measured plasma interleukin-(IL)1, tumor necrosis factor (TNF)α, and PMN-elastase, and serum neopterin and lysozyme in 107 patients with ME/CFS, 37 patients with chronic fatigue (CF), and 20 normal controls. The severity of ME/CFS was measured with the Fibromyalgia and Chronic Fatigue Syndrome (FF) Rating Scale.. Serum IL-1, TNFα, neopterin and lysozyme are significantly higher in patients with ME/CFS than in controls and CF patients. Plasma PMN-elastase is significantly higher in patients with ME/CFS than in controls and CF patients and higher in the latter than in controls. Increased IL-1 and TNFα are significantly correlated with fatigue, sadness, autonomic symptoms, and a flu-like malaise; neopterin is correlated with fatigue, autonomic symptoms, and a flu-like malaise; and increased PMN-elastase is correlated with concentration difficulties, failing memory and a subjective experience of infection.. The findings show that ME/CFS is characterized by low-grade inflammation and activation of CMI. The results suggest that characteristic symptoms of ME/CFS, such as fatigue, autonomic symptoms and a flu-like malaise, may be caused by inflammatory mediators, e.g. IL-1 and TNFα.

Topics: Adult; Biomarkers; Fatigue Syndrome, Chronic; Female; Humans; Immunity, Cellular; Inflammation; Interleukin-1; Leukocyte Elastase; Male; Middle Aged; Muramidase; Neopterin; Tumor Necrosis Factor-alpha

Most human transfusion recipients fail to make detectable alloantibodies to foreign RBC antigens ("nonresponders"). Herein, we use a murine model to test the hypothesis that nonresponders may be immunologically tolerant. FVB mice transfused with RBCs expressing transgenic human glycophorin A (hGPA) antigen in the absence of inflammation produced undetectable levels of anti-hGPA immunoglobulins, unlike those transfused in the presence of polyinosinic:polycytidylic acid-induced inflammation. Mice in the nonresponder group failed to produce anti-hGPA after subsequent transfusions in the presence of polyinosinic:polycytidylic acid, whereas anti-hGPA levels increased in the responder group. This tolerance was antigen specific, because nonresponders to hGPA produced alloantibodies to RBCs that expressed a different transgenic antigen. This tolerance was not an idiosyncrasy of the hGPA antigen nor of the recipient strain, because B10.BR mice transfused with membrane-bound hen egg lysozyme antigen-transgenic RBCs also demonstrated induced nonresponsiveness. These data demonstrate that RBCs transfused in the absence of inflammation can induce tolerance.

Topics: Analysis of Variance; Animals; Antibodies; Antigens; Enzyme-Linked Immunosorbent Assay; Erythrocyte Transfusion; Erythrocytes; Female; Glycophorins; Humans; Immune Tolerance; Inflammation; Isoantibodies; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Muramidase

Depression is characterized by inflammation and cell-mediated immune (CMI) activation and autoimmune reactions directed against a multitude of self-epitopes. There is evidence that the inflammatory response in depression causes dysfunctions in the metabolism of 5-HT, e.g. lowering the 5-HT precursor tryptophan, and upregulating 5-HT receptor mRNA. This study has been undertaken to examine autoimmune activity directed against 5-HT in relation to CMI activation and inflammation.. 5-HT antibodies were examined in major depressed patients (n=109) versus normal controls (n=35) in relation to serum neopterin and lysozyme, and plasma pro-inflammatory cytokines (PIC), i.e. interleukin-1 (IL-1) and tumor necrosis factor-α (TNFα). Severity of depression was assessed with the Hamilton Depression Rating Scale (HDRS) and severity of fatigue and somatic symptoms with the Fibromyalgia and Chronic Fatigue Syndrome (FF) Rating Scale.. The incidence of anti-5-HT antibody activity was significantly higher in depressed patients (54.1%), and in particular in those with melancholia (82.9%), than in controls (5.7%). Patients with positive 5-HT antibodies showed increased serum neopterin and lysozyme, and plasma TNFα and IL-1; higher scores on the HDRS and FF scales, and more somatic symptoms, including malaise and neurocognitive dysfunctions. There was a significant association between autoimmune activity to 5-HT and the number of previous depressive episodes.. The autoimmune reactions directed against 5-HT might play a role in the pathophysiology of depression and the onset of severe depression. The strong association between autoimmune activity against 5-HT and inflammation/CMI activation is explained by multiple, reciprocal pathways between these factors. Exposure to previous depressive episodes increases the incidence of autoimmune activity directed against 5-HT, which in turn may increase the likelihood to develop new depressive episodes. These findings suggest that sensitization (kindling) and staging of depression are in part based on progressive autoimmune responses.

Topics: Adult; Autoimmunity; Depressive Disorder; Depressive Disorder, Major; Female; Humans; Immunity, Cellular; Inflammation; Interleukin-1; Male; Middle Aged; Muramidase; Neopterin; Serotonin; Tumor Necrosis Factor-alpha

Subpopulations of pathogenic or nonpathogenic Th17 cells were reported to develop when presensitized CD4 cells were activated with their target Ag during polarization by either IL-23 or IL-6 and TGF-β, respectively. In this study, we generated two Th17 subpopulations by using a system in which naive CD4 cells from TCR transgenic mice specific to hen egg lysozyme (HEL) are polarized with IL-6/TGF-β and, concurrently, are activated either with HEL presented by APCs, or with anti-CD3/CD28 Abs. Only the former cells were pathogenic, inducing inflammation in eyes expressing HEL. Naive CD4 cells activated by the anti-CD3/CD28 Abs acquired pathogenicity, however, when cocultured with HEL/APC. Importantly, the naive CD4 cells did not acquire pathogenicity when cocultured with APCs stimulated with LPS or when separated from the HEL-presenting cells by a semipermeable membrane. Unlike with presensitized Th17, soluble IL-23 does not participate in pathogenicity acquisition by naive CD4 cells; no pathogenicity was induced by adding IL-23 to cultures activated with anti-CD3/CD28 Abs. Furthermore, Abs against IL-23 or IL-23R did not inhibit acquisition of pathogenicity in cultures of naive CD4 cells activated by HEL/APC. Our data thus show that, unlike presensitized CD4 cells, naive CD4 cells polarized toward Th17 phenotype acquire pathogenicity only by direct interaction with APCs presenting the Ag, with no apparent involvement of soluble IL-23. We suggest that the Th17 lymphocytes derived from naive CD4 cells participate in pathogenic and other immune processes, along with the IL-23-dependent Th17 cells.

Topics: Animals; Antigen-Presenting Cells; CD4-Positive T-Lymphocytes; Cell Communication; Cell Lineage; Cells, Cultured; Coculture Techniques; Dendritic Cells; Eye Diseases; Inflammation; Interleukin-23; Mice; Mice, Transgenic; Muramidase; Th17 Cells

Pathogenic bacteria colonize the airways of 30% to 40% of patients with COPD and cause approximately 50% of exacerbations. New strains of nontypeable Haemophilus influenzae (NTHI) and Moraxella catarrhalis are associated with exacerbations. Antimicrobial protein/peptides (AMPs) play important roles in innate lung defense against pathogens. To our knowledge, the changes in AMP baseline levels in respiratory secretions during bacterial colonization and exacerbation have not been described. The objective of this study was to elucidate the effects of the acquisition of a new strain of pathogenic bacteria on the airway levels of AMPs in patients with COPD.. One hundred fifty-three samples from 11 patients were selected from COPD sputum samples collected prospectively over 6 years. Samples were grouped as culture-negative (no pathogenic bacteria), colonization, and exacerbation due to new strains of NTHI and M catarrhalis. Levels of lysozyme, lactoferrin, LL-37, and secretory leukocyte protease inhibitor (SLPI) were measured by enzyme-linked immunosorbent assay and compared among groups by paired analysis.. Compared with baseline, sputum lysozyme levels were significantly lower during colonization and exacerbation by NTHI (P = .001 and P = .013, respectively) and M catarrhalis (P = .007 and P = .018, respectively); SLPI levels were lower with exacerbation due to NTHI and M catarrhalis (P = .002 and P = .004, respectively), and during colonization by M catarrhalis (P = 032). Lactoferrin levels did not change significantly; LL-37 levels were higher during exacerbation by NTHI and M catarrhalis (P = .001 and P = .018, respectively).. Acquisition of NTHI and M catarrhalis is associated with significant changes in airway levels of AMPs, with larger changes in exacerbation. Airway AMP levels are likely to be important in pathogen clearance and clinical outcomes of infection in COPD.

Topics: Aged; Aged, 80 and over; Antimicrobial Cationic Peptides; beta-Defensins; Cathelicidins; Disease Progression; Female; Haemophilus Infections; Haemophilus influenzae; Humans; Inflammation; Lactoferrin; Male; Middle Aged; Moraxella catarrhalis; Moraxellaceae Infections; Muramidase; Prospective Studies; Pulmonary Disease, Chronic Obstructive; Secretory Leukocyte Peptidase Inhibitor; Sputum

Nutritional fatty acids are known to have an impact on membrane lipid composition of body cells, including cells of the immune system, thus providing a link between dietary fatty acid uptake, inflammation and immunity. In this study we reveal the significance of macrophage membrane lipid composition on gene expression and cytokine synthesis thereby highlighting signal transduction processes, macrophage activation as well as macrophage defense mechanisms. Using RAW264.7 macrophages as a model system, we identified polyunsaturated fatty acids (PUFA) of both the n-3 and the n-6 family to down-regulate the synthesis of: (i) the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α; (ii) the co-stimulatory molecule CD86; as well as (iii) the antimicrobial polypeptide lysozyme. The action of the fatty acids partially depended on the activation status of the macrophages. It is particularly important to note that the anti-inflammatory action of the PUFA could also be seen in case of infection of RAW264.7 with viable microorganisms of the genera R. equi and P. aeruginosa. In summary, our data provide strong evidence that PUFA from both the n-3 and the n-6 family down-regulate inflammation processes in context of chronic infections caused by persistent pathogens.

Topics: Actinomycetales Infections; Animals; B7-2 Antigen; Cell Line; Dietary Fats; Down-Regulation; Fatty Acids, Unsaturated; Immunity, Innate; Inflammation; Interleukin-1beta; Interleukin-6; Macrophage Activation; Macrophages; Membrane Lipids; Mice; Muramidase; Myeloid Differentiation Factor 88; Pseudomonas aeruginosa; Pseudomonas Infections; Rhodococcus equi; Tumor Necrosis Factor-alpha

This paper reports the results of a study that aimed to assess whether liver functionality defined by liver activity index (LAI) is associated with inflammatory and immune parameters in blood and milk. LAI is an index including the average blood levels of albumin, lipoproteins and retinol-binding protein measured three times in the first month of lactation (at 5, 15 and 30 days in milk). The aim was to assess the relationship of this index with blood and udder immune and inflammatory status as a means of identifying as early as possible cows at risk of disease. The research was carried out using 10 multiparous Italian-Friesian dairy cows of average genetic merit. Cows were retrospectively ranked in three groups according the LAI level. Blood samplings were performed at different intervals before and after calving; quarter milk samples were taken only after calving with the same schedule as blood samples. Leucocytes, oxidative burst, blood lysozyme and N-acetyl-beta-d-glucosaminidase (NAGase) curves showed large overlapping among the three LAI group curves during the follow-up period. Four blood (complement, sialic acid, haptoglobin and reactive oxygen metabolites) and three milk (somatic cell count, lysozyme and NAGase) parameters showed larger and more consistent differences among LAI groups. Complement showed higher values and sialic acid showed lower values in high LAI group when compared with the other two LAI groups. Two other markers of inflammatory status (haptoglobin and reactive oxygen metabolites) showed the lowest values in high LAI cows. A consistent and significant reduction of milk NAGase and milk lysozyme in high LAI group was observed. The results suggest that cows with the highest liver functionality index have also the highest levels of some immune markers and the lowest levels for inflammatory markers at blood (already before calving) and mammary levels. Finally, cows with low LAI index, being more susceptible to metabolic and infectious diseases, should be carefully monitored to identify as early as possible the development of a disease.

Topics: Acetylglucosaminidase; Animals; Cattle; Cattle Diseases; Complement System Proteins; Female; Haptoglobins; Inflammation; Lactation; Leukocyte Count; Lipoproteins; Liver; Milk; Muramidase; N-Acetylneuraminic Acid; Reactive Oxygen Species; Retinol-Binding Proteins; Serum Albumin

One route to gain insight into the causes and consequences of ecological differentiation is to understand the underlying physiological mechanisms. We explored the relationships between immunological and oxidative status and investigated how birds cope physiologically with the effects of immune-derived oxidative damage. We successively implemented two experimental manipulations to alter physiological status in a model bird species: the homing pigeon (Columba livia). The first manipulation, an immune supplementation, was achieved by oral administration of lysozyme, a naturally occurring and non-specific antimicrobial enzyme. The second manipulation, an immune challenge, took the form of an injection with lipopolysaccharide, a bacterial endotoxin. Between groups of lysozyme-treated and control birds, we compared lipopolysaccharide-induced changes in reactive oxygen metabolites, total antioxidant capacity, haptoglobin, oxygen consumption, body mass and cloacal temperature. Lysozyme supplementation intensified the lipopolysaccharide-induced inflammatory response and generated short-term oxidative and metabolic costs. We identified significant interactions between immune supplementation and immune challenge in terms of reactive oxygen metabolites, haptoglobin and oxygen consumption. Our study provides alternative interpretations of differences in oxidative and immunological indices and demonstrates that these indices can also fluctuate and interact across very short time scales, reflecting something akin to current 'health status' or 'physiological condition'. These ephemeral effects highlight the need to broadly consider current physiological condition when drawing conclusions that relate physiology to ecology and evolution.

Topics: Animals; Antioxidants; Columbidae; Dietary Supplements; Ecological and Environmental Phenomena; Female; Inflammation; Lipopolysaccharides; Male; Models, Animal; Muramidase; Oxidants; Oxidative Stress; Oxygen Consumption; Reactive Oxygen Species

Recently reported lines of Th9 cells, producing IL-9 and IL-10, were generated by polarization with IL-4 and TGF-β and activation with Abs against CD3 and CD28. In this paper, we analyzed features of Th9 lines similarly polarized but activated by the "natural mode" (i.e., exposure of CD4 cells to their target Ag, hen egg lysozyme [HEL] and APCs). Main observations are the following: 1) both IL-9 and IL-10 were expressed by the line cells, but with strikingly different kinetics, with IL-9 being produced rapidly, reaching a peak on day 3 in culture and declining sharply thereafter, whereas IL-10 production increased gradually, resembling IL-4 and IL-17 production by their corresponding lineage cells; 2) reactivation of Th9, following expansion, triggered faster and higher production of both IL-9 and IL-10; 3) incubating Th9 cells in polarizing media specific for other phenotypes stimulated moderate levels of phenotype switching to Th1 or Th17 but a massive switching to Th2; 4) Th9 cells induced moderate inflammation in HEL-expressing recipient eyes but only when producing high levels of IL-9; and 5) IL-9-producing donor cells were detected in the blood of Th9 recipients but not in their inflamed eyes, suggesting that similar to findings in culture, exposure to HEL in these eyes arrested the IL-9 production in Th9 cells. Collectively, these data provide new information concerning Th9 cells and reveal their uniqueness, in particular with regard to the unusual production kinetics of IL-9 and the short retention of these cells in affected target tissues.

Topics: Animals; Cell Line; Chickens; Dose-Response Relationship, Immunologic; Epitopes, T-Lymphocyte; Eye Proteins; Inflammation; Inflammation Mediators; Interleukin-10; Interleukin-9; Kinetics; Mice; Mice, Inbred C57BL; Mice, Transgenic; Muramidase; Organ Specificity; Receptors, Antigen, T-Cell; T-Lymphocytes, Helper-Inducer; Time Factors

Vegetable oils (Vo) are an alternative to fish oil (Fo) in aquaculture feeds. This study aimed to evaluate the effect of dietary soybean oil (Vo diet), rich in linoleic acid, and of dietary fish oil (Fo diet) on the development of spinal deformities under bacterial lipopolysaccharide (LPS)-induced chronic inflammation conditions in Atlantic salmon, Salmo salar L. Fish [25 g body weight (BW)] were fed the experimental diets for 99 days. On day 47 of feeding (40 g BW), fish were subjected to four experimental regimes: (i) intramuscular injections with LPS, (ii) sham-injected phosphate-buffered saline (PBS), (iii) intraperitoneally injected commercial oil adjuvant vaccine, or (iv) no treatment. The fish continued under a common feeding regime in sea water for 165 more days. Body weight was temporarily higher in the Vo group than in the Fo group prior to immunization and was also affected by the type of immunization. At the end of the trial, no differences were seen between the dietary groups. The overall prevalence of spinal deformities was approximately 14% at the end of the experiment. The Vo diet affected vertebral shape but did not induce spinal deformities. In groups injected with LPS and PBS, spinal deformities ranged between 21% and 38%, diet independent. Deformed vertebrae were located at or in proximity to the injection point. Assessment of inflammatory markers revealed high levels of plasma prostaglandin E₂ (PGE₂) in the Vo-fed and LPS-injected groups, suggesting an inflammatory response to LPS. Cyclooxigenase 2 (COX-2) mRNA expression in bone was higher in fish fed Fo compared to Vo-fed fish. Gene expression of immunoglobulin M (IgM) was up-regulated in bone of all LPS-injected groups irrespective of dietary oil. In conclusion, the study suggests that Vo is not a risk factor for the development of inflammation-related spinal deformities. At the same time, we found evidence that localized injection-related processes could trigger the development of vertebral body malformations.

Topics: Animal Feed; Animals; Bone and Bones; Diet; Dietary Fats; Fatty Acids; Fish Diseases; Inflammation; Irritants; Lipopolysaccharides; Muramidase; Radiography; Salmo salar; Spinal Diseases; Spine

In spite of the many studies on protein modifications by reactive species, knowledge about the products resulting from the oxidation of protein-aromatic residues, including protein-derived radicals and their stable products, remains limited. Here, we compared the oxidative modifications promoted by peroxynitrite and myeloperoxidase/hydrogen peroxide/nitrite in two model proteins, ribonuclease (6Tyr) and lysozyme (3Tyr/6Trp). The formation of protein-derived radicals and products was higher at pH 5.4 and 7.4 for myeloperoxidase and peroxynitrite, respectively. The main product was 3-nitro-Tyr for both proteins and oxidants. Lysozyme rendered similar yields of nitro-Trp, particularly when oxidized by peroxynitrite. Hydroxylated and dimerized products of Trp and Tyr were also produced, but in lower yields. Localization of the main modified residues indicates that peroxynitrite decomposes to radicals within the proteins behaving less specifically than myeloperoxidase. Nitrogen dioxide is emphasized as an important protein modifier.

Topics: Animals; Cattle; Chickens; Chromatography, High Pressure Liquid; Egg White; Free Radicals; Hydrogen Peroxide; Inflammation; Muramidase; Peroxidase; Peroxynitrous Acid; Pronase; Proteins; Ribonucleases; Spectrometry, Mass, Electrospray Ionization; Spleen; Tyrosine

The study aimed to describe the patterns and density of early tracheal colonization among intubated patients and to correlate colonization status with levels of antimicrobial peptides and inflammatory cytokines.. The was a prospective cohort study.. The study was conducted in medical and cardiovascular intensive care units of a tertiary referral hospital.. Seventy-four adult patients admitted between March 2003 and May 2006 were recruited for the study.. Tracheal aspirates were collected daily for the first 4 days of intubation using standardized, sterile technique and sent for quantitative culture and cytokines, lactoferrin and lysozyme measurements.. The mean acute physiology and chronic health evaluation (APACHE II) score in this cohort was 24 +/- 7. Proportion of subjects colonized by any microorganism increased over the first 4 days of intubation (47%, 60%, 70%, 70%, P = .08), but density of colonization for bacteria or yeast did not change significantly. No known risk factors predicted tracheal colonization on day 1 of intubation. Several patterns of colonization were observed (persistent, transient, new colonization, and clearance of initial colonization).The most common organisms cultured were Candida albicans and coagulase-negative Staphylococcus. Levels of cytokines, lactoferrin, or lysozyme did not change over time and were not correlated with tracheal colonization status. Four subjects (6%) had ventilator-associated pneumonia.. The density of tracheal colonization did not change significantly over the first 4 days of intubation in medical intensive care unit patients. There was no correlation between tracheal colonization and the levels of antimicrobial peptides or cytokines. Several different patterns of colonization may have to be considered while planning interventions to reduce airway colonization.

Topics: Adult; APACHE; Candidiasis; Case-Control Studies; Colony Count, Microbial; Cross Infection; Cytokines; Female; Humans; Inflammation; Intensive Care Units; Intubation, Intratracheal; Lactoferrin; Logistic Models; Male; Middle Aged; Multivariate Analysis; Muramidase; Pneumonia, Ventilator-Associated; Prospective Studies; Respiration, Artificial; Respiratory Mucosa; Risk Factors; Staphylococcal Infections; Statistics, Nonparametric; Suction; Time Factors; Trachea

Whether the wound macrophage is a key regulatory inflammatory cell type in skin repair has been a matter of debate. A transgenic mouse model mediating inducible macrophage depletion during skin repair has not been used to date to address this question. Here, we specifically rendered the monocyte/macrophage leukocyte lineage sensitive to diphtheria toxin by expressing the lysozyme M promoter-driven, Cre-mediated excision of a transcriptional STOP cassette from the simian DT receptor gene in mice (lysM-Cre/DTR). Application of diphtheria toxin to lysM-Cre/DTR mice led to a rapid reduction in both skin tissue and wound macrophage numbers at sites of injury. Macrophage-depleted mice revealed a severely impaired wound morphology and delayed healing. In the absence of macrophages, wounds were re-populated by large numbers of neutrophils. Accordingly, macrophage-reduced wound tissues exhibited the increased and prolonged persistence of macrophage inflammatory protein-2, macrophage chemoattractant protein-1, interleukin-1beta, and cyclooxygenase-2, paralleled by unaltered levels of bioactive transforming growth factor-beta1. Altered expression patterns of vascular endothelial growth factor on macrophage reduction were associated with a disturbed neo-vascularization at the wound site. Impaired wounds revealed a loss of myofibroblast differentiation and wound contraction. Our data in the use of lysM-Cre/DTR mice emphasize the pivotal function of wound macrophages in the integration of inflammation and cellular movements at the wound site to enable efficient skin repair.

Topics: Animals; Blotting, Western; Cell Line; Cell Lineage; Diphtheria Toxin; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Humans; Immunohistochemistry; Inflammation; Keratinocytes; Macrophages; Mice; Mice, Transgenic; Muramidase; Neovascularization, Physiologic; Poisons; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor beta; Wound Healing

Degradable hydrogels have been extensively used in biomedical applications such as drug delivery, and recent interest has grown in hydrogels that degrade in recognition of a cellular response. This contribution describes a poly(ethylene glycol) (PEG) hydrogel platform with human neutrophil elastase (HNE) sensitive peptide cross-links formed using thiol-ene photopolymerization rendering the gel degradable at sites of inflammation. Further, protein therapeutics can be physically entrapped within the network and selectively released upon exposure to HNE. HNE-responsive hydrogels exhibited surface erosion where the degradation kinetics was influenced by changes in peptide k(cat), concentration of HNE, and concentration of peptide within the gel. Using this platform, we were able to achieve controlled, zero-order release of bovine serum albumin (BSA) in the presence of HNE, and release was arrested in the absence of HNE. To further exploit the advantages of surface eroding delivery systems, a smaller protein (carbonic anhydrase) was delivered at the same rate as BSA and only dependent on gel formulation and environmental conditions. Also, protein release was predicted from a 3-layered hydrogel device using mass loss data. Lastly, the bioactivity of lysozyme was maintained above 90% following the exposure to thiol-ene photopolymerization conditions.

Topics: Animals; Biocompatible Materials; Cattle; Drug Delivery Systems; Enzymes; Humans; Hydrogels; Inflammation; Kinetics; Leukocyte Elastase; Light; Muramidase; Polyethylene Glycols; Proteins; Serum Albumin; Sulfhydryl Compounds; Surface Properties

Both Th1 and Th17 T cell subsets can mediate inflammation, but the kinetics of the pathogenic processes mediated by these two subsets have not been investigated. Using an experimental system in which TCR-transgenic Th1 or Th17 cells specific for hen egg lysozyme induce ocular inflammation in recipient mice expressing eye-restricted hen egg lysozyme, we found important differences in the in vivo behavior of these two subsets. Th1 cells initially proliferated considerably faster and invaded the eye more quickly than their Th17 counterparts, but then disappeared rapidly. By contrast, Th17 cells accumulated and remained the majority of the infiltrating CD4(+) cells in the eye for as long as 25 days after transfer, mediating more long-lasting pathological changes. Unlike Th1, Th17 cells were highly resistant to restimulation-induced apoptosis, a major pathway by which autoimmune and chronically restimulated Th1 cells are eliminated. Th17 cells had reduced Fas ligand production and resistance to Fas-induced apoptosis, relative to Th1 cells, despite similar surface expression of Fas. Th17-induced ocular inflammation also differed from Th1-induced inflammation by consisting of more neutrophils, whereas Th1-induced disease had higher proportions of CD8 cells. Taken together, our data show that pathogenic processes triggered by Th17 lag behind those induced by Th1, but then persist remarkably longer, apparently due to the relative resistance of Th17 cells to restimulation-induced cell death. The long-lasting inflammation induced by Th17 cells is in accord with these cells being involved in chronic conditions in humans.

Topics: Adoptive Transfer; Animals; Apoptosis; Autoimmune Diseases; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Division; Eye; Fas Ligand Protein; fas Receptor; Flow Cytometry; Inflammation; Interleukin-17; Mice; Mice, Transgenic; Muramidase; T-Lymphocyte Subsets; Th1 Cells

Th1 and Th17 cells are characterized by their expression of IFN-gamma or IL-17, respectively. The finding of Th cells producing both IL-17 and IFN-gamma suggested, however, that certain Th cells may modify their selective cytokine expression. In this study, we examined changes in cytokine expression in an experimental system in which polarized Th1 or Th17 cells specific against hen egg lysozyme induce ocular inflammation in recipient mice expressing hen egg lysozyme in their eyes. Whereas only IFN-gamma was expressed in eyes of Th1 recipient mice, substantial proportions of donor cells expressed IFN-gamma or both IFN-gamma and IL-17 in Th17 recipient eyes. The possibility that nonpolarized cells in Th17 preparations were responsible for expression of IFN-gamma or IFN-gamma/IL-17 in Th17 recipient eyes was contradicted by the finding that the proportions of such cells were larger in recipients of Th17 preparations with 20-25% nonpolarized cells than in recipients of 35-40% preparations. Moreover, whereas incubation in vitro of Th1 cells with Th17-polarizing mixture had no effect on their phenotype, incubation of Th17 with Th1-polarizing mixture, or in the absence of cytokines, converted most of these cells into IFN-gamma or IFN-gamma/IL-17-expressing cells. In addition, Th17 incubated with the Th1 mixture expressed T-bet, whereas no ROR-gamma t was detected in Th1 incubated with Th17 mixture. Thus, polarized Th1 cells retain their phenotype in the tested systems, whereas Th17 may switch to express IFN-gamma or IFN-gamma/IL-17 following activation in the absence of cytokines, or exposure to certain cytokine milieus at the inflammation site or in culture.

Topics: Animals; Eye Diseases; Flow Cytometry; Immunophenotyping; Inflammation; Interferon-gamma; Interleukin-17; Lymphocyte Activation; Mice; Mice, Transgenic; Muramidase; Phenotype; Receptors, Antigen, T-Cell; Reverse Transcriptase Polymerase Chain Reaction; T-Lymphocyte Subsets; Th1 Cells

Observations that dendritic cells (DCs) constitutively enter afferent lymphatic vessels in many organs and that DCs in some tissues, such as the lung, turnover rapidly in the steady state have led to the concept that a major fraction of lymph node DCs are derived from migratory DCs that enter the lymph node through upstream afferent lymphatic vessels. We used the lysozyme M-Cre reporter mouse strain to assess the relationship of lymph node and nonlymphoid organ DCs. Our findings challenge the idea that a substantial proportion of lymph node DCs derive from the upstream tissue during homeostasis. Instead, our analysis suggests that nonlymphoid organ DCs comprise a major population of DCs within lymph nodes only after introduction of an inflammatory stimulus.

Topics: Animals; Cell Movement; Dendritic Cells; Genes, Reporter; Green Fluorescent Proteins; Homeostasis; Inflammation; Lung; Lymph Nodes; Lymphatic Vessels; Macrophages; Mice; Mice, Inbred C57BL; Mice, Transgenic; Muramidase; Spleen

There is increasing evidence of systemic inflammation in patients with chronic obstructive pulmonary disease (COPD), but there is very little information on the development of systemic inflammation in smokers without severe airway symptoms. In this longitudinal study, we examined whether smokers with mild or no airway symptoms develop signs of systemic inflammation by assessing inflammatory markers in blood over a 6-year period.. Forty smokers and 28 male never-smokers were investigated in 1995 (year 0) and 6 years later (year 6). At year 6, 11 smokers had stopped smoking (quitters); these subjects were analysed as a separate group. At year 0 and 6, we measured serum levels of myeloperoxidase (MPO), lysozyme and human neutrophil lipocalin (HNL), regarded as markers of activity in neutrophils plus monocyte-lineage cells, monocyte-lineage cells only and neutrophils only.. All systemic markers of inflammation (MPO, HNL and lysozyme) were significantly higher in smokers than in never smokers at year 6. For MPO alone, smokers only displayed a unique pattern compared with the other groups; the concentration of MPO in blood increased among smokers during the 6-year period, and this increase was statistically significant compared with that observed in never-smokers. Even though quitters did not display any clear change in MPO, we observed a statistically significant negative correlation between the change in blood MPO and the duration of smoking cessation in this group. For HNL and lysozyme, the changes over time were similar in smokers and never-smokers, with no statistically significant difference compared with quitters.. This study provides evidence that male smokers without severe airway symptoms develop an increasing systemic inflammation during a 6-year period. The study forwards both direct and indirect evidence that MPO may be an early marker of this systemic inflammation. However, our study also forwards indirect evidence that ongoing tobacco smoking may "drive" the level of systemic HNL and lysozyme. The origin of the increased MPO and its value as an easily measured predictor for future COPD deserves to be further evaluated.

Topics: Acute-Phase Proteins; Aged; Biomarkers; Body Height; Body Mass Index; Follow-Up Studies; Forced Expiratory Volume; Humans; Inflammation; Lipocalin-2; Lipocalins; Male; Muramidase; Peroxidase; Proto-Oncogene Proteins; Pulmonary Diffusing Capacity; Respiratory Function Tests; Smoking; Smoking Cessation; Time Factors

Aging may be accompanied by a low grade chronic up-regulation of inflammatory mediators. A variety of endogenous locally released mediators as well as inflammatory cells have been reported in the human oral cavity. The aim of this investigation was to determine the presence of different classes of inflammatory mediators in human saliva and correlate the levels with age.. Unstimulated whole buccal salivary samples were obtained in the morning from 94 healthy volunteers within 30 minutes after waking. None of the participants had taken aspirin in the week prior to the saliva collection. Lysozyme activity, eicosanoid levels (prostaglandin E(2) and leukotriene B(4)) and MMP-9 activity were measured. The antimicrobial activity (lysozyme activity) was not correlated with age whereas PGE(2) levels were markedly correlated with age (r = 0.29; P<0.05; n = 56). Saliva from healthy subjects (< or =40 years) compared with data derived from older volunteers (>40 years) demonstrated a significant increase in the mean values for PGE(2) and MMP-9 activity with age. In addition, significant correlations were observed between LTB(4) and PGE(2) (r = 0.28; P<0.05; n = 56) and between LTB(4) levels and MMP-9 activity in smokers (r = 0.78; P<0.001; n = 15).. The presence of significant levels and activity of inflammatory mediators in saliva suggests that the oral cavity of healthy subjects may be in a constant low state of inflammation associated with age.

Topics: Adolescent; Adult; Aged; Aging; Dinoprostone; Female; Humans; Inflammation; Inflammation Mediators; Leukotriene B4; Matrix Metalloproteinase 9; Middle Aged; Mouth; Muramidase; Saliva

The effect of somatic cell count (SCC) and milk fraction on milk composition, distribution of cell populations, and mRNA expression of various inflammatory parameters was studied. Therefore, quarter milk samples were defined as cisternal (C), first 400 g of alveolar (A1), and remaining alveolar milk (A2) during the course of milking. Quarters were assigned to 4 groups according to their total SCC: 1) <12 x 10(3)/mL, 2) 12 to 100 x 10(3)/mL, 3) 100 to 350 x 10(3)/mL, and 4) >350 x 10(3)/mL. Milk constituents of interest were SCC, fat, protein, lactose sodium, and chloride ions as well as electrical conductivity. Cell populations were classified into lymphocytes, macrophages, and neutrophils (PMN). The mRNA expression of the inflammatory factors tumor necrosis factor-alpha, interleukin-1beta, cyclooxygenase-2, lactoferrin, and lysozyme was measured via real-time, quantitative reverse transcription PCR. Somatic cell count decreased from highest levels in C to lowest levels in A1 and increased thereafter to A2 in all groups. Fat content increased from C to A2 and with increasing SCC level. Lactose decreased with increasing SCC level but remained unchanged during milking. Concentrations of sodium and chloride, and electrical conductivity increased with increasing SCC but were higher in C than in A1 and A2. Protein was not affected by milk fraction or SCC level. The distribution of leukocytes was dramatically influenced by milk fraction and SCC. Lymphocytes were the dominating cell population in group 1, but the proportion of lymphocytes was low in groups 2, 3, and 4. Macrophage proportion was highest in group 2 and decreased in groups 3 and 4, whereas that of PMN increased from group 2 to 4. The content of macrophages decreased during milking in all SCC groups whereas that of PMN increased. The proportion of lymphocytes was not affected by milk fraction. The mRNA expression of all inflammatory factors showed an increase with increasing SCC but minor changes occurred during milking. In conclusion, milk fraction and SCC level have a crucial influence on the distribution of leukocyte populations and several milk constituents. The surprisingly high content of lymphocytes and concomitantly low mRNA expression of inflammatory factors in quarters with SCC <12 x 10(3)/mL indicates a different and possibly reduced readiness of the immune system to respond to invading pathogens.

Topics: Animals; Cattle; Cattle Diseases; Cell Count; Cyclooxygenase 2; Female; Inflammation; Interleukin-1; Lactoferrin; Leukocyte Count; Lymphocyte Count; Macrophages; Milk; Muramidase; Neutrophils; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Necrosis Factor-alpha

The anti-inflammatory effect of total phenolics from Laggera alata (TPLA) was evaluated with various in vivo models of both acute and chronic inflammations. In the acute inflammation tests, TPLA inhibited significantly xylene-induced mouse ear oedema, carrageenan-induced rat paw oedema and acetic acid-induced mouse vascular permeability. In the carrageenan-induced rat pleurisy model, TPLA significantly suppressed inflammatory exudate and leukocyte migration, reduced the serum levels of lysozyme (LZM) and malondialdehyde (MDA), increased the serum levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX), and also decreased the contents of total protein, nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in the pleural exudates. In the chronic inflammation experiment, TPLA inhibited significantly cotton pellet-induced rat granuloma. These results indicated that TPLA possesses potent anti-inflammatory activity on acute and chronic inflammation models. Its anti-inflammatory mechanisms are probably associated with the inhibition of prostaglandin formation, the influence on the antioxidant systems, and the suppression of LZM release. Furthermore, the total phenolic content of Laggera alata and its main component type was quantified, and its principle components were isolated and authenticated. Acute toxicity studies revealed that TPLA up to an oral dose of 8.5 g/kg body weight was almost nontoxic in mice.

Topics: Acute Disease; Animals; Asteraceae; Capillary Permeability; Carrageenan; Chronic Disease; Dexamethasone; Ear, External; Edema; Glutathione Peroxidase; Inflammation; Male; Malondialdehyde; Mice; Mice, Inbred ICR; Muramidase; Nitric Oxide; Plant Extracts; Pleurisy; Quinic Acid; Rats; Rats, Sprague-Dawley; Superoxide Dismutase; Xylenes

Most alloantigens on transfused red blood cells (RBCs) are weakly immunogenic, with only a 2 to 6 percent overall immunization rate even in patients receiving multiple transfusions. Although recipient genetics may contribute to responder and/or nonresponder status, in most cases HLA type does not predict humoral response to RBC antigens. In contrast, rates of alloimmunization do correspond to the underlying disease status of transfusion recipients, suggesting that acquired host factors may play an important role. In this context, it was hypothesized that the inflammatory status of a transfusion recipient would influence immunization to transfused RBCs.. A novel murine model for alloimmunization to RBC antigens was developed with the mHEL mouse, which expresses hen egg lysozyme (HEL) as a model blood group antigen. Leukoreduced mHEL RBCs were transfused into wild-type recipient mice, and anti-HEL responses were monitored. To test the stated hypothesis, some recipient animals were injected with polyinosinic polycytidylic acid (poly(I:C)), a synthetic double-stranded RNA molecule that induces viral-like inflammation.. Similar to the immunogenicity of most RBC antigens in humans, transfusion of mHEL RBCs into uninflamed mice was only a weak immunogen. In contrast, poly(I:C)-treated mice had a significant increase in both the frequency and the magnitude of alloimmunization to the mHEL antigen.. These findings demonstrate that recipient inflammation with poly(I:C) significantly enhances humoral immunization to transfused alloantigens in a murine model. Moreover, these data suggest that the inflammatory status of human transfusion recipients may regulate the immunogenicity of transfused RBCs.

Topics: Animals; Antibody Formation; Crosses, Genetic; Erythrocyte Transfusion; Erythrocytes; Immunization; Immunoglobulin G; Inflammation; Isoantigens; Leukocyte Reduction Procedures; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Mice, Transgenic; Models, Animal; Muramidase; Poly I-C

Recent studies claim a central role for Toll-like receptor (TLR) ligands in stimulating autoimmune disease by activation of antigen-presenting cells in the target organ, but it is unclear if and how TLR ligands reach target organs. Most evidence comes from rodent models, and it is uncertain whether this principle holds in primates. Here we identify which cells contain peptidoglycan (PGN) in multiple sclerosis brain and in two nonhuman primate experimental autoimmune encephalomyelitis (EAE) models with different disease courses: acute (rhesus monkey) versus chronic disease (marmoset). Because persistence of TLR ligands in the central nervous system might be consequential for disease progression, we also determined the expression of two major PGN-degrading enzymes, ie, lysozyme and N-acetylmuramyl-l-alanine amidase. Distinct phagocyte subsets, including granulocytes, macrophages, and dendritic cells, contained PGN in the brain and coexpressed the inflammatory cytokine interleukin-12. The number of phagocytes carrying PGN increased in acute and chronic EAE compared with control animals, with the highest number of PGN-containing cells in acute EAE brain. Lytic enzymes were scarcely expressed in monkey and multiple sclerosis brain, favoring PGN persistence. PGN stimulated interleukin-12p70 release by leukocytes from all three primate species. The presence of PGN in the inflamed brain may have major implications because TLR2/Nod ligation potentially promotes inflammation and disease progression.

Topics: Adult; Aged; Aged, 80 and over; Animals; Brain; Callithrix; Demyelinating Diseases; Encephalomyelitis, Autoimmune, Experimental; Female; Humans; Inflammation; Leukocytes, Mononuclear; Ligands; Macaca mulatta; Male; Middle Aged; Muramidase; N-Acetylmuramoyl-L-alanine Amidase; Peptidoglycan; Phagocytes; Solubility; Staphylococcus aureus; Toll-Like Receptors

Virgin olive oil (VOO) compared with fish oil (FO) and evening primrose oil (PO) on the ability of stimulated leukocytes to produce inflammatory mediators was investigated in rats. Weaned Wistar rats were fed a basal diet (BD) (2% by weight of corn oil) or diets containing 15% by weight of VOO, PO, or FO. After 8 weeks, glycogen-elicited peritoneal polymorphonuclear leukocytes, mainly neutrophils, were isolated. The calcium-ionophore stimulated neutrophils (2.5 x 10(6) cells/mL) obtained from rats fed the different oils produced a higher release of lysosomal enzymes (beta-glucuronidase, lysozyme, and myeloperoxidase [MPO]) compared with those fed BD. The production of reactive oxygen species (ROS) in response to the stimulant, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), by neutrophils from the VOO group (15.44 nmol of O(2)(-) and 6.56 nmol of H(2)O(2)) was similar to the BD group (12.01 nmol O(2)(-) and 8.49 nmol H(2)O(2)) and significantly lower than the PO (20.90 nmol O(2)(-) and 10.84 nmol H(2)O(2)) and FO (20.93 nmol O(2)(-) and 12.79 nmol H(2)O(2)) groups. The cyclooxygenase-derived eicosanoid production was reduced by the lipid enrichment of the diets. Whereas the generation of prostaglandin E(2) (PGE(2)) was significantly decreased in VOO (5.40 ng/mL), PO (4.95 ng/mL), and FO (1.44 ng/mL) groups compared with BD (8.19 ng/mL), thromboxane B(2) (TXB(2)) reduction was especially significant in neutrophils from the FO diet group (14.67 ng/mL compared with 26.69 ng/mL from BD). These experimental data suggest that FO and PO, as well as VOO, could be considered a valuable strategy in preventing the generation of some inflammatory mediators.

Topics: Animals; Arachidonic Acid; Calcimycin; Dietary Fats, Unsaturated; Dinoprostone; Eicosanoids; Fatty Acids; Fatty Acids, Essential; Fish Oils; gamma-Linolenic Acid; Glucuronidase; Glycogen; Hydrogen Peroxide; Inflammation; Linoleic Acid; Linoleic Acids; Lysosomes; Male; Muramidase; Neutrophils; Oenothera biennis; Oleic Acid; Olive Oil; Peritoneum; Peroxidase; Plant Oils; Rats; Rats, Wistar; Reactive Oxygen Species; Superoxides; Tetradecanoylphorbol Acetate; Thromboxane B2

Incomplete deletion of KRN T cells that recognize the ubiquitously expressed self-antigen glucose-6-phosphate-isomerase (GPI) initiates an anti-GPI autoimmune cascade in K/BxN mice resulting in a humorally mediated arthritis. Transgenic (Tg) expression of a KRN T cell receptor (TCR) agonist under the major histocompatibility complex class II promoter resulted in thymic deletion with loss of anti-GPI T and B cell responses and attenuated arthritis course. However, double Tg mice succumbed to systemic autoimmunity with multiorgan inflammation and autoantibody production. Extensive thymic deletion resulted in lymphopenia and elimination of CD4+ CD25+ regulatory T cells (Tregs), but spared some CD4+ T cells expressing endogenous TCR, which oligoclonally expanded in the periphery. Disease was transferred by these T cells and prevented by cotransfer of CD4+ CD25+ Tregs. Moreover, we extended our findings to another TCR system (anti-hen egg lysozyme [HEL] TCR/HEL mice) where similarly extensive thymic deletion also resulted in disease. Thus, our studies demonstrated that central tolerance can paradoxically result in systemic autoimmunity through differential susceptibility of Tregs and autoreactive T cells to thymic deletion. Therefore, too little or too much negative selection to a self-antigen can result in systemic autoimmunity and disease.

Topics: Animals; Antigens, CD; Base Sequence; CD4-Positive T-Lymphocytes; Chickens; DNA Primers; Enzyme-Linked Immunosorbent Assay; Erythrocytes; Inflammation; Lymphocyte Depletion; Mice; Mice, Transgenic; Muramidase; Mutagenesis; Polymerase Chain Reaction; Receptors, Interleukin-2; Spleen; T-Lymphocytes; Thymectomy

More than 70 years ago, Alexander Fleming discovered lysozyme and proposed that nonpathogenic bacteria fail to cause disease because they are very susceptible to destruction by lysozyme, an enzyme that is one of the principal proteins of phagocytes. Although much has been learned about the effects of lysozyme in vitro, its biological role in vivo has not been determined. We examined transgenic mice deficient in lysozyme M after challenge by the normally nonpathogenic and highly lysozyme-sensitive bacterium Micrococcus luteus. Despite partial compensation by newly expressed lysozyme P in macrophages, lysozyme M-deficient mice developed much more severe lesions than wild-type mice. The tissue injury was due to the failure of lysozyme M-deficient mice to inactivate peptidoglycan, resulting in an intense and prolonged inflammatory response. Our data indicate that tissue injury is normally limited by prompt degradation of bacterial macromolecules that trigger innate immunity and inflammation.

Topics: Animals; Bacterial Infections; Disease Susceptibility; Gene Expression; Green Fluorescent Proteins; Inflammation; Luminescent Proteins; Macrophages; Mice; Mice, Inbred C57BL; Mice, Transgenic; Micrococcus luteus; Muramidase; Neutrophils; Peptidoglycan; Recombinant Fusion Proteins

We have previously shown that transgenic (Tg) mice expressing either soluble or membrane-bound hen egg lysozyme (sHEL or mHEL, respectively) under control of the alphaA-crystallin promoter develop tolerance due to thymic expression of minuscule amounts of HEL. To further address the mechanisms by which this tolerance develops, we mated these two lines of Tg mice with the 3A9 line of HEL-specific TCR Tg mice, to produce double-Tg mice. Both lines of double-Tg mice showed deletion of HEL-specific T cells, demonstrated by reduction in numbers of these cells in the thymus and periphery, as well as by reduced proliferative response to HEL in vitro. In addition, the actual deletional process in thymi of the double-Tg mice was visualized in situ by the TUNEL assay and measured by binding of Annexin V. Notably, the apoptosis localized mainly in the thymic medulla, in line with the finding that the populations showing deletion and increased Annexin V binding consisted mainly of single- and double-positive thymocytes. Interestingly, the thymic deletional effect of sHEL was superior to that of mHEL in contrast to the opposite differential tolerogenic effects of these HEL forms on B cells specific to this Ag. Analysis of bone marrow chimeras indicates that both forms of HEL are produced by irradiation-resistant thymic stromal cells and the data suggest that sHEL is more effective in deleting 3A9 T cells due mainly to its higher accessibility to cross-presentation by dendritic APC.

Topics: Animals; Antigen Presentation; Antigens; Apoptosis; Autoantigens; Clonal Deletion; Dose-Response Relationship, Immunologic; Epithelial Cells; Eye; Eye Proteins; Immunophenotyping; In Situ Nick-End Labeling; Inflammation; Lymphocyte Activation; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Transgenic; Muramidase; Self Tolerance; Solubility; Stromal Cells; T-Lymphocyte Subsets; Thymus Gland

Class A scavenger receptors (SR-A) have several proposed functions that could impact atherosclerosis and inflammatory processes. To define the function of SR-A in vivo, we created C57BL/6 transgenic mice that expressed bovine SR-A under the control of the restricted macrophage promoter, lysozyme (lyso-bSR-A). bSR-A mRNA was present in cultured peritoneal macrophages of transgenic mice and tissues that contain significant macrophages including spleen, lung, and ileum. Functional overexpression of SR-A was demonstrated in peritoneal macrophages both by augmented cholesterol ester deposition in response to AcLDL and enhanced adhesion in transgenic mice compared with nontransgenic littermates. To determine whether macrophage-specific expression of bSR-A regulated inflammatory responses, granulomas were generated by subcutaneous injection of carrageenan. Granuloma size was significantly increased in lyso-bSR-A transgenic mice compared with wild-type littermates [421 +/- 51 mg (n = 11) vs. 127 +/- 22 mg (n = 10), P < 0.001]. However, the larger granulomas in lyso-bSR-A transgenic mice were only associated with an increase in unesterified cholesterol, and not cholesterol esters. Furthermore, granulomas from transgenic mice had an increase in the number of macrophages within the tissue.Therefore, macrophage expression of bSR-A increased presence of this cell type in granulomas without enhancing the deposition of cholesterol esters, consistent with a role of the adhesive property of the protein.

Topics: Animals; Carrageenan; Cholesterol; Disease Models, Animal; Gene Expression; Granuloma; Inflammation; Lipoproteins, LDL; Macrophages, Peritoneal; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Transgenic; Muramidase; Promoter Regions, Genetic; Receptors, Immunologic; Receptors, Lipoprotein; Receptors, Scavenger; Scavenger Receptors, Class A; Scavenger Receptors, Class B

The anti-inflammatory activities of the isolated flavonoids, quercetin 3-O-methyl ether (1), kaempferol (2), and quercetin (3), of Rhamnus nakaharai, and anthraquinone, frangulin B (4), of Rhamnus formosana, were assessed in vitro by determining their inhibitory effects on the chemical mediators released from mast cells, neutrophils, macrophages, and microglial cells. Compounds 1 - 3 strongly inhibited the release of beta-glucuronidase and lysozyme from rat neutrophils stimulated with formyl-Met-Leu-Phe/cytochalasin B (fMLP/CB). Compound 1 strongly inhibited superoxide anion formation in fMLP/CB or phorbol 12-myristate 13-acetate (PMA)-stimulated rat neutrophils. Compound 1 exhibited potent inhibitory effect on tumor-necrosis factor-alpha ( TNF-alpha) formation in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells while 1 and 4 showed potent inhibitory effects on TNF-alpha formation in LPS/IFN-gamma (interferon-gamma)-stimulated murine microglial cell lines N9.

Topics: Animals; Anthraquinones; Anti-Inflammatory Agents, Non-Steroidal; Cell Degranulation; Flavonoids; Glucuronidase; Inflammation; Kaempferols; Macrophages; Male; Microglia; Molecular Structure; Muramidase; Neutrophils; Phytotherapy; Plant Bark; Plant Preparations; Quercetin; Rats; Rhamnus; Structure-Activity Relationship

During their final differentiation or maturation, dendritic cells (DCs) redistribute their major histocompatibility complex (MHC) class II products from intracellular compartments to the plasma membrane. Using cells arrested in the immature state, we now find that DCs also regulate the initial intracellular formation of immunogenic MHC class II-peptide complexes. Immature DCs internalize the protein antigen, hen egg lysozyme (HEL), into late endosomes and lysosomes rich in MHC class II molecules. There, despite extensive colocalization of HEL protein and MHC class II products, MHC class II-peptide complexes do not form unless the DCs are exposed to inflammatory mediators such as tumor necrosis factor alpha, CD40 ligand, or lipoplolysaccharide. The control of T cell receptor (TCR) ligand formation was observed using the C4H3 monoclonal antibody to detect MHC class II-HEL peptide complexes by flow cytometry and confocal microscopy, and with HEL-specific 3A9 transgenic T cells to detect downregulation of the TCR upon MHC-peptide encounter. Even the binding of preprocessed HEL peptide to MHC class II is blocked in immature DCs, including the formation of C4H3 epitope in MHC class II compartments, suggesting an arrest to antigen presentation at the peptide-loading step, rather than an enhanced degradation of MHC class II-peptide complexes at the cell surface, as described in previous work. Therefore, the capacity of late endosomes and lysosomes to produce MHC class II-peptide complexes can be strictly controlled during DC differentiation, helping to coordinate antigen acquisition and inflammatory stimuli with formation of TCR ligands. The increased ability of maturing DCs to load MHC class II molecules with antigenic cargo contributes to the >100-fold enhancement of the subsequent primary immune response observed when immature and mature DCs are compared as immune adjuvants in culture and in mice.

Topics: Animals; Antigen Presentation; CD40 Antigens; CD40 Ligand; Cell Differentiation; Dendritic Cells; Female; Histocompatibility Antigens Class II; Inflammation; Injections, Subcutaneous; Ligands; Lysosomes; Membrane Glycoproteins; Mice; Mice, Inbred C3H; Mice, Inbred CBA; Muramidase; Peptides

Transgenic (Tg) mice expressing a foreign Ag, hen egg lysozyme (HEL), under control of the alphaA-crystallin promoter ("HEL-Tg" mice) develop immunotolerance to HEL attributed to the expression of HEL in their thymus. In this paper we analyzed the immune response in double (Dbl)-Tg mice generated by mating the HEL-Tg mice with Tg mice that express HEL Abs on their B cells ("Ig-Tg" mice). The B cell compartment of the Dbl-Tg mice was unaffected by the HEL presence and was essentially identical to that of the Ig-Tg mice. A partial breakdown of tolerance was seen in the T cell response to HEL of the Dbl-Tg mice, i.e., their lymphocyte proliferative response against HEL was remarkably higher than that of the HEL-Tg mice. T-lymphocytes of both Dbl-Tg and Ig-Tg mice responded to HEL at concentrations drastically lower than those found stimulatory to lymphocytes of the wild-type controls. Cell mixing experiments demonstrated that 1) the lymphocyte response against low concentrations of HEL is due to the exceedingly efficient Ag presenting capacity of the Ab expressing B cells and 2) breakdown of tolerance in Dbl-Tg mice can also be attributed to the APC capacity of B cells, that sensitize in vivo and stimulate in vitro populations of T cells with low affinity toward HEL, assumed to be escapees of thymic deletion. These results thus indicate that T cell tolerance can be partially overcome by the highly potent Ag presenting capacity of Ab expressing B cells.

Topics: Animals; Antigen Presentation; Antigen-Presenting Cells; Antigens, Surface; Autoantigens; B-Lymphocyte Subsets; Cytokines; Immunoglobulins; Inflammation; Lens, Crystalline; Mice; Mice, Inbred C57BL; Mice, Transgenic; Muramidase; Receptors, Antigen, B-Cell; Self Tolerance; T-Lymphocyte Subsets

The mechanism was studied of the anti-inflammatory effect of oral zinc (114 mg/kg/day of elemental metal, given for 14 days) on the development of the carrageenan-induced paw oedema of the rat, and the impact of in vivo treatment on the activity of neutrophils isolated from the blood of inflamed and non-inflamed animals. The effects of the in vitro incubation with the metal on either non-inflamed or inflamed neutrophils coming from zinc-untreated rats were also examined. It was found that the administration of oral zinc inhibited markedly the process of ex vivo adhesion of the cells obtained from the inflamed rats (an observation confirmed by the in vitro experiments). In vitro release of lysozyme and superoxide anion productions were measured: in the absence of zinc, the 30' of pre-incubation carried out before stimulating with PMA did not influence the cell's reactivity of the non-inflamed neutrophils. It was, on the contrary, capable of significantly reducing that of the inflamed ones. As a consequence, it is quite difficult to properly interpret the data obtained studying the activity of the cells exposed to the metal in vitro.

Topics: Animals; Anti-Inflammatory Agents; Copper; Female; In Vitro Techniques; Inflammation; Liver; Muramidase; Neutrophils; Platelet Adhesiveness; Rats; Rats, Sprague-Dawley; Superoxides; Tetradecanoylphorbol Acetate; Zinc

The aim of the present study was to examine the relationships between the ventilation rate and the type of ventilation system, on the one hand, and objective nasal measures, on the other.. A standardized investigation, including acoustic rhinometry and nasal lavage, was performed in the school environment. All 279 school personnel working in the main buildings of 12 randomly selected primary schools in the municipality of Uppsala were invited, and 234 (84%) participated. The dimensions of the nasal cavity were measured with acoustic rhinometry. Eosinophil cationic protein (ECP), myeloperoxidase (MPO), lysozyme, and albumin were analyzed in the lavage fluid. The air exchange rate and the room temperature were measured in the classrooms. Relationships between nasal symptoms, nasal patency, and the concentration of biomarkers, on the one hand, and the type of ventilation system, the air exchange rate, and the temperature, on the other, were analyzed by both crude bivariate analysis and multiple regression models, controlling for the type of ventilation, the air exchange rate, room temperature, age, gender, smoking, atopy, and the urban vicinity of the school.. A lower degree of nasal patency as measured by acoustic rhinometry and increased levels of ECP and lysozyme in nasal lavage were associated with a lower air exchange rate in the schools. Although mechanically ventilated schools had higher air exchange rates, they were associated with more nasal symptoms, and nasal mucosal swelling and with increased lavage levels of ECP and lysozyme as compared with schools with natural ventilation only. In contrast, 12 subjects working in a school with mechanical displacement ventilation had more patent noses and lower levels of inflammatory markers as compared with the personnel in schools with natural ventilation only.. Our results indicate that both a low air exchange rate and mechanical ventilation systems based on dilution can be associated with reduced nasal patency and an inflammatory biomarker response of the nasal mucosa among school personnel. The only school with sufficient ventilation according to the current Swedish recommendations had a displacement system and the fewest signs of nasal reactions among the personnel.

Topics: Albumins; Analysis of Variance; Biomarkers; Blood Proteins; Cross-Sectional Studies; Eosinophil Granule Proteins; Female; Follow-Up Studies; Humans; Inflammation; Linear Models; Male; Middle Aged; Muramidase; Nasal Lavage Fluid; Nasal Obstruction; Occupational Diseases; Peroxidase; Ribonucleases; Schools; Sweden; Ventilation

Lysozyme is an ubiquitous enzyme found in most biological secretions and leukocytes. This study was aimed at investigating its interaction with other inflammatory mediators on mucosa surfaces, particularly the complement system. Lysozyme has been shown in our present study, to inhibit the haemolytic activity of serum complement in a dose-dependent fashion, when tested within the levels present in normal and inflamed breast-milk samples, and other mucosal secretions. This represents a new anti-inflammatory action of lysozyme in relation to the serum complement, and the exact mode of the interaction need further studies.

Topics: Animals; Complement Activation; Complement Hemolytic Activity Assay; Erythrocytes; Female; Hemolysis; Humans; Immunity, Mucosal; In Vitro Techniques; Inflammation; Inflammation Mediators; Milk, Human; Muramidase; Sheep

We found that granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) had different effects on the apoptosis of human mature neutrophils induced by anti-Fas antibody. GM-CSF suppressed this process and neutrophils retained their functions of superoxide production and enzyme release against invasion of microorganisms. In contrast, G-CSF had a weaker effect on the anti-Fas antibody-induced neutrophil apoptosis than GM-CSF, with neutrophil function suppressed in proportion to the apoptosis. GM-CSF produced by satellite cells at the inflammatory site may inhibit apoptosis and the neutrophils may maintain their primed functions to kill the invading microorganisms. G-CSF produced by fibroblasts or endothelial cells may work in another fashion by increasing the number of neutrophils more effectively than GM-CSF. GM-CSF and G-CSF may exert different effects on neutrophils at the inflammatory sites in the human body.

Topics: Antibodies, Monoclonal; Apoptosis; fas Receptor; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation; Muramidase; Neutrophils; Recombinant Proteins; Superoxides

T cell receptor (TCR) transgenic mice specific for hen egg lysozyme (HEL) were crossed with mice expressing HEL on the thyroid epithelium, on pancreatic islet beta cells, or systemically. Depending on the pattern of HEL expression, deletion of double-positive thymocytes ranged from minimal to complete, and peripheral CD4 cells exhibited graded reduction in TCR expression, in vitro responsiveness, and in vivo helper ability. CD4 cells were least tolerant in TCR/thyroid-HEL and TCR/islet-HEL mice, which developed an extensive lymphocytic thyroiditis or insulitis that nevertheless did not eliminate HEL-expressing endocrine cells. Autoreactive CD4 clones thus escape the thymus under a range of circumstances, retain sufficient function to initiate subclinical autoimmune inflammation when self-antigens are concentrated in the thyroid or pancreas, and may regulate progression of subclinical inflammation to destructive autoimmune disease.

Topics: Animals; CD4-Positive T-Lymphocytes; Cell Movement; Chickens; Enzyme Activation; Epitopes, T-Lymphocyte; Immune Tolerance; Immunophenotyping; Inflammation; Islets of Langerhans; Lymphocyte Count; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Muramidase; Necrosis; Organ Specificity; Receptors, Antigen, T-Cell, alpha-beta; Thyroid Gland; Thyroiditis, Autoimmune

The CD19 cell surface molecule regulates signal transduction events critical for B lymphocyte development and humoral immunity. Increasing the density of CD19 expression renders B lymphocytes hyper-responsive to transmembrane signals, and transgenic mice that overexpress CD19 have increased levels of autoantibodies. The role of CD19 in tolerance regulation and autoantibody generation was therefore examined by crossing mice that overexpress a human CD19 transgene with transgenic mice expressing a model autoantigen (soluble hen egg lysozyme, sHEL) and high-affinity HEL-specific IgMa and IgDa (IgHEL) antigen receptors. In this model of peripheral tolerance, B cells in sHEL/IgHEL double-transgenic mice are functionally anergic and do not produce autoantibodies. However, it was found that overexpression of CD19 in sHEL/IgHEL double-transgenic mice resulted in a breakdown of peripheral tolerance and the production of anti-HEL antibodies at levels similar to those observed in IgHEL mice lacking the sHEL autoantigen. Therefore, altered signaling thresholds due to CD19 overexpression resulted in the breakdown of peripheral tolerance. Thus, CD19 overexpression shifts the balance between tolerance and immunity to autoimmunity by augmenting antigen receptor signaling.

Topics: Animals; Antibody Specificity; Antigens, CD19; Autoantibodies; Autoantigens; Autoimmune Diseases; Autoimmunity; B-Lymphocyte Subsets; Chickens; Clonal Anergy; Complement C3d; Crosses, Genetic; Humans; Immune Tolerance; Immunoglobulin D; Immunoglobulin M; Inflammation; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Transgenic; Muramidase; Receptors, Antigen, B-Cell; Receptors, Complement 3d; Signal Transduction

Human N-acetylmuramyl-L-alanine amidase (EC 3.5.1.28) degrades peptidoglycan, a major component of bacterial cell walls with potent pro-inflammatory cytokine-inducing properties. We postulate that degradation of peptidoglycan by N-acetylmuramyl-L-alanine amidase is important for the inactivation of inflammatory peptidoglycan products in human tissues. The inflammatory activities of peptidoglycan digested by lysozyme and/or amidase were investigated using two properties of peptidoglycan: its capacity to induce the release of the inflammatory cytokines IL-1, IL-6 and TNF-alpha in vivo and in vitro and its capacity to induce arthritis in Lewis rats. The results show that after subsequent treatment with both lysozyme and amidase, the peptidoglycan products were unable to induce arthritis in Lewis rats. The production of pro-inflammatory cytokines in mice after intravenous injection of cell wall fragments was lower after in vitro degradation of the cell wall fragments by amidase. These in vivo results were confirmed with whole blood assays in which the production of pro-inflammatory cytokines was measured after stimulation with lysozyme- and amidase-treated peptidoglycan. The results show that human N-acetylmuramyl-L-alanine amidase possesses an enzymatic activity capable of inactivating inflammatory peptidoglycan by lowering its cytokine-inducing properties.

Topics: Animals; Carbohydrate Sequence; Female; Humans; Inflammation; Interleukin-1; Interleukin-6; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Muramidase; N-Acetylmuramoyl-L-alanine Amidase; Peptidoglycan; Rats; Rats, Inbred Lew; Tumor Necrosis Factor-alpha

Dendritic cells (DCs) have emerged as the dominant antigen-presenting cells (APCs) of the lung, playing a vital role in the induction of cell-mediated immunity to inhaled antigens. We have previously demonstrated that an airway challenge with the soluble antigen hen egg lysozyme yields rapid acquisition of specific antigen-presenting cell activity by purified pulmonary DCs and a cell-mediated immune response in the lung upon secondary challenge. To examine how a particulate antigen leads to a cell-mediated response in vivo, graded concentrations of heat-killed Listeria (HKL) were injected intratracheally into Lewis rats. The bacteria were rapidly ingested by lung macrophages and polymorphonuclear leukocytes. The ability of purified pulmonary DCs pulsed in vivo by an airway challenge with HKL to subsequently stimulate HKL-specific responses ex vivo showed a threshold response, requiring a dose in excess of 10(9) organisms/rat. By contrast, all dosages of HKL yielded specific sensitization of lymphocytes in the draining bilar nodes. Pulmonary DCs purified from rats after a secondary in vivo airway challenge with HKL at day 14 were ineffective antigen-presenting cells except at high dosages of antigen. The generation of cell-mediated pulmonary inflammation paralleled the antigen-presenting cell activity of pulmonary DCs and was observed only at high antigen dosages. Hen egg lysozyme immobilized onto polystyrene beads and injected intratracheally yielded comparable results to those observed with HKL. We suggest that a pulmonary cellular immune response is generated to an inhaled particulate antigen when the protective phagocytic capacities of the lung are exceeded and antigen is able to interact directly with interstitial DCs. The diversion of particulate antigens by pulmonary phagocytes may help to limit undesirable pulmonary inflammation while allowing the generation of antigen-specific immune lymphocytes in vivo.

Topics: Animals; Antigens, Bacterial; Bronchoalveolar Lavage Fluid; Dendritic Cells; Female; Immunity, Cellular; Inflammation; Listeria monocytogenes; Lung; Macrophages, Alveolar; Muramidase; Neutrophils; Phagocytes; Phagocytosis; Rats; Rats, Inbred Lew; T-Lymphocytes

Although many cytokines have been implicated in the development and persistence of inflammatory immune responses, it is unknown if any of these are important in inflammatory acne. This study investigated the production of the proinflammatory cytokines interleukin-8 (IL-8), IL-1 beta, and tumor necrosis factor alpha (TNF-alpha) by human monocytic cell lines, ThP-1 and U937, and by freshly isolated peripheral blood mononuclear cells from acne patients. Both Propionibacterium acnes and supernatants obtained from 72-h P. acnes cultures could induce significant concentrations of IL-1 beta, TNF-alpha, and IL-8 by both cell lines and by peripheral blood mononuclear cells as determined by enzyme-linked immunosorbent assay. There was no significant difference between acne and non-acne subjects. Endotoxin quantification and addition of polymyxin B to assays indicated no lipopolysaccharide (LPS) contamination. P. acnes supernatant was fractionated into components with molecular weights of < 3,000, < 10,000, and < 30,000 and assayed for the ability to induce IL-8 and TNF production in ThP-1 cells. Nearly 90% of the original activity was found in the < 30,000-molecular-weight fraction, 50% was in the < 10,000-molecular-weight fraction, and only 15% remained in the < 3,000-molecular-weight fraction. The effluent from the < 3,000-molecular-weight fraction contained about 70% activity, indicating that the inducing factor was not retained in the membrane. Incubation of P. acnes supernatant with various concentrations of mutanolysin or lysozyme resulted in a loss of 60% of the original activity. The addition of jimson lectin, which binds peptidoglycan, resulted in a loss of 70% of the activity in a dose-response manner, whereas peanut lectin had little or no effect on the activity. Heating of the P. acnes supernatant to 65 degrees C also had no effect on the activity. Blocking of CD14, a receptor for both LPS and peptidoglycan, reduced cytokine production by > 50%, suggesting that the soluble stimulating factor may be a secreted form of peptidoglycan-polysaccharide.

Topics: Acne Vulgaris; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Cell Wall; Cells, Cultured; Chronic Disease; Endopeptidases; Endotoxins; Humans; In Vitro Techniques; Inflammation; Interleukin-1; Interleukin-8; Keratinocytes; Lipopolysaccharide Receptors; Monocytes; Muramidase; Propionibacterium acnes; Tumor Necrosis Factor-alpha

The aggregation of non-serotypable Haemophilus influenzae (NTHI) by whole saliva from patients with chronic obstructive lung disease (COLD) was investigated. Significant differences were observed between salivary aggregating activity of a control and COLD population (P < 0.001). Saliva from patients less prone to acute exacerbations had a greater capacity to aggregate bacteria compared with saliva from patients with a predilection to infection. The mechanism of saliva-mediated aggregation of NTHI was investigated and shown to be related to lysozyme content. Lysozyme activity in saliva was measured by the turbidimetric technique and results showed that patients with chronic bronchitis had increased levels of salivary lysozyme, with a subpopulation within the non-infection-prone group having greater amounts. A significant difference was observed in salivary lysozyme between controls and non-infection-prone (P < 0.005) and infection-prone (P < 0.05) patients, respectively: the non-infection-prone patients having significantly (P < 0.005) more than the infection-prone patients. There was significant correlation (r = 0.742, P < 0.001) between salivary aggregation of NTHI and lysozyme activity. Chromatographically purified human lysozyme had a similar aggregation profile to that of saliva. There was no difference in serum and saliva lactoferrin concentrations between groups, but there was a significant increase (P < 0.05) in serum lysozyme concentration in the non-infection-prone group. This study suggests that the level of salivary lysozyme derived from macrophages may play an important role in determining resistance or susceptibility to acute bronchitis.

Topics: Acute Disease; Adult; Aged; Bronchitis; Chronic Disease; Communicable Diseases; Female; Haemophilus influenzae; Humans; Inflammation; Lactoferrin; Lung Diseases, Obstructive; Macrophages; Male; Middle Aged; Monocytes; Muramidase; Neutrophils; Saliva; Salivation

The hypothesis that intermittent claudication initiates a systemic inflammatory response was investigated by studying the effect of exercise on markers of neutrophil activation and vascular permeability in 25 claudicants and 10 controls. Urinary albumin excretion, previously demonstrated to reflect vascular permeability, increased significantly after exercise in claudicants and was associated with decreased neutrophil filterability and increased serum lysozyme activity. No similar exercise-induced changes were seen in controls or in claudicants after successful arterial bypass surgery. These results suggest that intermittent claudication is associated with potentially deleterious systemic manifestations that are surgically reversible.

Topics: Aged; Aged, 80 and over; Albuminuria; Capillary Permeability; Creatinine; Exercise; Female; Hemofiltration; Humans; Inflammation; Intermittent Claudication; Male; Middle Aged; Muramidase; Neutrophils

Topics: Amyloidosis; Animals; Glycoproteins; Inflammation; Mice; Muramidase

Exudate and uterine flushings were collected at either 30, 60, 120 or 240 mins after intrauterine infusions of Streptococcus zooepidemicus in genitally normal mares during oestrus. Uteri were also flushed without prior induction of endometritis. Protein concentrations in exudate and flushings increased with time and exudate pH decreased with time; the pH of flushings did not alter. Lysozyme and lactate dehydrogenase were present in flushings from non-infected uteri, but concentrations increased with time after infection. Immunoreactive prostaglandin E2 was undetectable before infection, but concentrations rose after infection. No neutrophils were present in non-infected flushings but, by 30 mins, there were significant (P less than 0.01) neutrophil numbers in exudate and flushings; thereafter numbers increased, particularly in exudate. Acute endometritis resembled acute inflammation at other sites in the horse and a significant response had occurred by 30 mins after experimental infection.

Topics: Animals; Chemotaxis, Leukocyte; Endometritis; Female; Horse Diseases; Horses; Hydrogen-Ion Concentration; Inflammation; L-Lactate Dehydrogenase; Leukocyte Count; Muramidase; Neutrophils; Prostaglandins E; Proteins; Uterus

Association of platelets and neutrophils is frequently observed within thrombi or inflammatory sites. Interactions between these two cell populations have been reported for the production of several mediators of inflammation such as hydrogen peroxides or leukotrienes. Another potential mediator of thrombosis and inflammation is paf-acether, which is synthesized by activated platelets and neutrophils. Since platelets form and release large amounts of the paf-acether precursor lyso paf-acether, platelet and neutrophil cooperation for paf-acether biosynthesis was investigated. Purified human neutrophils (4 x 10(6)/ml) stimulated by opsonized zymosan (ZC, 1 mg/ml) formed 4.5 +/- 2.5 ng/ml paf-acether. Human washed platelets (3 x 10(8)/ml) stimulated with thrombin (1 IU/ml) formed 0.60 +/- 0.43 ng/ml paf-acether. Platelets and neutrophils, incubated together and both stimulated by their specific agonist, formed more than twice as much paf-acether as did platelets and neutrophils separately (10.90 +/- 4.25 ng/ml, n = 6, P less than .001). The formation of lyso paf-acether and the release of lysozyme and LDH were unchanged under the cooperation conditions. The formation of paf-acether almost doubled (10.24 +/- 3.81 ng/ml paf-acether vs. 5.30 +/- 2.23, P less than .05, n = 4) when ZC-stimulated neutrophils were incubated with supernatants from thrombin-stimulated platelets as well as with synthetic lyso paf-acether. Extracted and purified lyso paf-acether from thrombin-stimulated platelets led to an increase of biosynthesis of paf-acether by neutrophils (13.86 +/- 2.26 ng/ml paf-acether vs. 5.76 +/- 0.38, P less than .05, n = 3). These results indicate that a cooperation between platelets and neutrophils exists for paf-acether formation. The phenomenon depends on a platelet-derived soluble factor, possibly lyso paf-acether. This cell-to-cell interaction is of interest since paf-acether is formed by and acting on platelets and neutrophils and represents a molecular basis for potent amplification of inflammatory reactions.

Topics: Animals; Blood Platelets; Cell Communication; Female; Humans; Inflammation; L-Lactate Dehydrogenase; Male; Muramidase; Neutrophils; Platelet Activating Factor; Rabbits; Thrombin; Zymosan

Topics: Humans; Inflammation; Muramidase; Neoplasms; Neutrophils

During the process of extravasation, monocytes transiently adhere to capillary endothelium and subsequently with a variety of extracellular matrix components. These early interactions are likely to serve as modifiers of transcriptional activity and serve to prime monocytes for rapid synthesis of mediators. We have previously reported that adherence to plastic rapidly induced or down-regulated steady-state mRNA levels of a number of monocyte inflammatory mediator genes. We now report that adherence to surfaces pretreated with fibronectin resulted in TNF-alpha and CSF-1 mRNA levels approximating adherence to plastic, whereas adherence to fibronectin, fibronectin/anti-fibronectin complexes, or collagen resulted in markedly decreased levels of CSF-1 induction and lysozyme down-regulation. In contrast, monocyte adherence to collagen induced the highest sustained levels of TNF-alpha expression. PMA, but not the chemotactic factor FMLP, stimulated non-adherent monocytes to express c-fos and TNF-alpha and down-regulate lysozyme mRNA. Although all donors responded to adherence, several failed to produce CSF-1 mRNA after PMA stimulation in the non-adherent state. These data demonstrate that monocyte mediator expression may be more dependent on the selectivity of signals induced by adherence to different substrates than the initial chemotactic response.

Topics: Antigens, Surface; Cell Adhesion; Cell Adhesion Molecules; Cells, Cultured; Colony-Stimulating Factors; Culture Media; Extracellular Matrix; Gene Expression Regulation; Humans; Inflammation; Monocytes; Muramidase; Protein Synthesis Inhibitors; Tetradecanoylphorbol Acetate; Tumor Necrosis Factor-alpha

We have examined the effects of carrageenan inflammation activated by silica particles on implanted femoral-head cartilage. The injection of silica particles modifies the inflammatory response, inducing a prolonged polymorphonuclear leucocyte infiltration and low exudate volume. This modification in the inflammatory response resulted in an accelerated loss of proteoglycan by the implanted femoral heads. Examination of exudate interleukin 1 (IL-1) concentrations directly by a lymphocyte-activating assay and indirectly by measuring the acute-phase response indicated that the increased loss was not due to increased levels of IL-1. High levels of lactate dehydrogenase would suggest that the increase cartilage breakdown is due to dying polymorphonuclear leucocytes releasing their intracellular enzymes. It is concluded that injection of silica together with carrageenan produces a pathological environment similar to that seen in human septic arthritis.

Topics: Animals; Carrageenan; Cartilage; Inflammation; Interleukin-1; L-Lactate Dehydrogenase; Leukocytes, Mononuclear; Male; Muramidase; Neutrophils; Osteochondritis; Proteoglycans; Rats; Silicon Dioxide

beta-Adrenergic blockade has been clinically associated with vascular occlusion. Because neutrophil activation, either alone or in concert with other blood cells, can induce vaso-occlusion by aggregation, leukoembolization, and inflammatory damage to vascular endothelium, we studied the effect of beta-adrenergic blockade on neutrophil activation in vitro and on microvascular integrity in vivo. beta-Adrenergic antagonists, propranolol and alprenolol, could induce dose-related and stereospecific activation of human neutrophils resulting in granulocyte aggregation and migration. Propranolol produced de novo aggregation and also enhanced responses to chemotaxins, N-formyl-methionyl-leucyl-phenylalanine and C5a. Lysosomal exocytosis was unaffected by beta blockers. In partial explanation of the enhancement of granulocyte activation by beta blockade, we observed that propranolol produced enhanced expression and affinity of granulocyte surface receptors for tritiated N-formyl-methionyl-leucyl-phenylalanine. This enhanced activation of human granulocytes by beta blockers was manifest in vivo as augmented inflammatory dermal edema. Enhanced granulocyte activation by beta blockade may induce microvascular disruption and subsequent tissue inflammation.

Topics: Alprenolol; Animals; Cell Adhesion; Cell Aggregation; Cell Movement; Chemotaxis, Leukocyte; Glucuronidase; Humans; In Vitro Techniques; Inflammation; Lysosomes; Muramidase; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Propranolol; Rabbits; Receptors, Formyl Peptide; Receptors, Immunologic

We have examined sixty-seven surgically removed allograft kidneys to identify the different leukocyte subsets of interstitial infiltration and the early vascular lesions which characterize renal allograft rejection. Histochemical and immunohistochemical results (mouse monoclonal antibodies anti: Leu 1, Leu 3a-3b, Leu 7, Leu 2a, OK Ia-Dr, OKB2, Leu M1, Leu M3; rabbit heteroclonal antibodies anti -: IgA, IgG, IgM, C3, fibrinogen, lysozyme; lectins-ABC: RCA, WGA, UEA) and routine histochemical staining have shown an increase of T-helper and T-activated lymphocyte subsets in acute rejection. Neutrophilic leucocytes were present in hyperacute rejection; macrophages were also noted. In chronic rejection, several lymphocyte subsets, in different ratios, were identified. Monocyte/macrophage leukocytes were the most abundant cell population. IgA deposits were noted on tubular epithelia in hyperacute and chronic rejections. IgM deposits were observed in vascular walls in chronic rejection. C3 and fibrinogen deposits were seen in glomerular capillaries and in arterial walls, although in different ratios, in all cases of renal allograft rejections. We have generally seen weak reactions to IgG deposits. Histochemical analysis of lectin receptors has given different results according to the type of rejection considered. In hyperacute rejection, receptors for WGA were found both on glomerular endothelial cells and on the tubular brush border. In the latter, receptors for RCA were also found. In acute rejection, receptors for UEA and WGA were found in a lower number of cases of acute vascular rejection. In acute cellular rejection, receptors for RCA, UEA and WGA were recognized in tubular epithelia. In acute vascular rejection, as well as in chronic rejection, only receptors for WGA were present on tubular epithelia and on capillary loop endothelium. The use of anti-human lysozyme antibodies has yielded the following results: in acute and hyperacute rejection, when renal failure occurred, we saw a high ratio of lysozyme, either coarsely granular or diffuse in the proximal tubular epithelia. Lysozyme was found in myelocyte/macrophage cells within capillary loops and arterial walls, when acute necrotizing vasculitis was present. In acute rejection, proximal tubular cells were lysozyme-negative or lysozyme-positive only segmentally, especially when obliterative vasculitis by fibrointimal proliferation was present and renal function progressively failed. In most of the chron

Topics: Acute Disease; Graft Rejection; Humans; Immunoglobulins; Inflammation; Kidney; Kidney Transplantation; Lectins; Lymphocytes; Muramidase; Plant Lectins; Receptors, Mitogen; Staining and Labeling; Vascular Diseases; Wheat Germ Agglutinins

An 11-year-old patient with inflammatory fibrous histiocytoma of the iliac bone is presented. In addition to routine histopathological procedures the lesion was studied by enzyme-histochemical and immuno-histochemical methods. The results of acid phosphatase, alpha-naphthyl-acetate esterase and intracytoplasmic muramidase demonstration in tumour cells supported the histiocytic nature of the presented case.

Topics: Bone Neoplasms; Child; Diagnosis, Differential; Female; Histiocytoma, Benign Fibrous; Histocytochemistry; Humans; Ilium; Immunoenzyme Techniques; Inflammation; Muramidase

Streptococci and streptococcal cell wall fragments induce arthritis in rats, with the severity and duration depending on the capacity of the cells or cell fragments to resist degradation by tissue enzymes. Their phlogogenic effects are apparently related to their ability to activate the alternate complement pathway (ACP). The in vitro activation of the ACP by lysozyme-treated cells and cell walls of group A, B, and D streptococci suggests that both rat and human lysozyme can modulate this activity, i.e., increasing it, decreasing it, or doing both in that order. The effects of the lysozymes also correlated with the degree to which they can unmask the aminosugar-reducing groups detectable in a given amount of cell wall, which suggests that partial depolymerization of the cell wall is critical for ACP activation. The effects of mutanolysin and C phage lysin on ACP activation were found to be correlated with their action on streptococcal cell walls. Neuraminidase had relatively little effect on ACP activation by most streptococcal strains tested. We conclude that the participation of tissue enzymes, including but not necessarily limited to lysozyme, is an important determinant for the clinical arthritis induced by group A, B, or D streptococci. Experimental arthritis induced in rats with whole (or disrupted) streptococci may depend both on the capacities of the cell walls to activate the ACP and on the capacities of the host tissue enzymes to modulate this activation. Great severity and long durations of the disease were determined by the capacity of the enzymes to degrade cell wall antigens to a degree sufficient to ensure efficient activation of the ACP without completely degrading the material so that it no longer activates complement. In this model, the limited resistance of group B peptidoglycan to lysozyme was a critical pathogenic factor.

Topics: Animals; Cell Wall; Chemotactic Factors; Complement Activation; Complement Fixation Tests; Complement Pathway, Alternative; Endopeptidases; Enterococcus faecalis; Humans; Inflammation; Muramidase; Neuraminidase; Polysaccharides, Bacterial; Rats; Streptococcus; Streptococcus agalactiae; Streptococcus pyogenes

After biosynthetic radiolabeling, polypeptides secreted by macrophages were analyzed by electrophoresis on polyacrylamide gels in the presence of sodium dodecyl sulfate and fluorography. Comparative studies of various macrophage populations (inflammatory macrophages, primed and activated macrophages, resident macrophages) showed that secretory activity is elaborately controlled according to macrophage developmental and functional state. When young macrophages activated to cytotoxicity against tumor cells by an appropriate eliciting agent and culture in the presence of lipopolysaccharide were compared to mature resident macrophages devoid of antitumor activity, the following changes were observed in the protein secretion phenotype: (1) New polypeptides are expressed: the intensity of four bands was increased more than five-fold on one-dimensional gels and 17 new spots were detected on two-dimensional gels. (2) The production of several major polypeptides is repressed: six bands were strongly decreased on one-dimensional gels, and 11 major spots (or groups of spots) were lost on two-dimensional gels. (3) The charge microheterogeneity pattern exhibited by several polypeptides on two-dimensional gels is shifted towards less acidic species.

Topics: Animals; Apolipoproteins E; Cytotoxicity, Immunologic; Female; Glycoproteins; Inflammation; Isoelectric Point; Macrophage Activation; Macrophages; Mice; Molecular Weight; Muramidase; Proteins; Time Factors

Tissue macrophages produce several proteins of the complement system. The mechanisms that regulate this process are poorly understood. The established ability of certain prostaglandins to influence macrophage secretory activity suggests that these lipid mediators may also modulate complement production (CP). Using the guinea pig peritoneal macrophages, we determined the effects of selected prostaglandins on in vitro CP and found that PGE2 inhibited production of complement proteins but not lysozyme; the response of elicited and resident peritoneal cells to PGE2 was identical; and PGE1, PGF2 alpha, and PGI2 had no detectable effect. PGE2 may contribute to regulation of CP in vivo.

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Complement System Proteins; Dose-Response Relationship, Drug; Female; Guinea Pigs; Inflammation; Kinetics; Macrophages; Male; Muramidase; Prostaglandins

Bovine phagocytic cells (polymorphonuclear granulocytes, blood monocytes and alveolar macrophages) were treated in vitro with homogeneous, recombinant DNA produced bovine alpha-1 interferon (IFN-alpha 1). The effects seen comprised of enhanced bacterial uptake by all three cell types and increased Fc receptor activity in alveolar macrophages, inhibition of both directed and random migration of monocytes and polymorphs, increased enzyme release or inactivation, increased hydrogen peroxide generation, and decreased superoxide anion release by alveolar macrophages and polymorphonuclear leukocytes. These effects were dose- and time-dependent, the kinetics varying for the different cell types.

Topics: Animals; Cattle; Cells, Cultured; DNA, Recombinant; Glucuronidase; Hydrogen Peroxide; Inflammation; Interferon Type I; Macrophages; Monocytes; Muramidase; Neutrophils; Phagocytosis; Staphylococcus aureus; Superoxides

Indomethacin (0.5 mg/100 g b.w./day) and chloramphenicol (0.5 mg or 15 mg/100 g b.w./day) were tested for their anti-inflammatory effects on 7th day carrageenan-induced granuloma formation. Neither of the drugs modified granuloma or pouch wall weight but they decreased the exudate and the cluster of dead cells. Indomethacin and chloramphenicol decreased glucosamine in the dead cell granuloma fraction and increased the level of collagen in the pouch wall. The drugs differed in their inhibitory effect on lysozyme and prostaglandin E2 accumulation in the exudate. The increase in collagen was related to a drop in the level of prostaglandin E2 which seems to regulate collagen deposition in the granuloma. However, the prostaglandin E2-lysozyme correlation--which was only significant with chloramphenicol--suggests a mode of action for chloramphenicol different from that of indomethacin. Chloramphenicol could act by a myelodepressive and/or chemotactic effect. The effects of chloramphenicol on the macrophages are discussed.

Topics: Animals; Body Weight; Carrageenan; Chloramphenicol; Collagen; Glycosaminoglycans; Granuloma; Indomethacin; Inflammation; Leukocyte Count; Male; Muramidase; Organ Size; Prostaglandins E; Rats; Rats, Inbred Strains

Using the peroxidase antiperoxidase (PAP) method, lysozyme (LZM) was shown to exist in normal, reactive and neoplastic cells belonging to the mononuclear phagocyte system (MPS), but was not detected in histiocytosis X cells. Immunostaining for cytoplasmic LZM by the PAP method is useful for identification of mononuclear phagocytes and for diagnosis of the diseases in which these cells participate.

Topics: Granulomatosis with Polyangiitis; Histiocytes; Hodgkin Disease; Humans; Immunoenzyme Techniques; Inflammation; Leukemia, Myeloid; Lymphadenitis; Lymphatic Diseases; Lymphoma, Large B-Cell, Diffuse; Mononuclear Phagocyte System; Muramidase; Neoplasms

Mouse inflammatory macrophages from C57BL/6N mice were fused with BALB/c mouse-derived myeloma cells (the CANS series). The hybrids in the early period after cell fusion (8 weeks) showed no macrophage functions (chemotaxis, EA and EAC rosette-forming abilities, phagocytosis or lysozyme production). EA rosette-forming ability was observed when these hybrids were treated with trypsin, whereas other macrophage functions were not. After prolonged culture, the hybrids (12 clones of 13 randomly selected) showed all the macrophage functions along with chromosome loss. Myeloma cell functions (kappa light chain production) were found in the young hybrids soon after cell fusion but were absent in the aged hybrids. These results indicated that reexpression of macrophage properties, except for EA rosette-forming abilities, takes place after the loss of chromosomes or genes repressing the expression of macrophage functions.

Topics: Animals; Cell Line; Chemotactic Factors; Chemotaxis; Hybrid Cells; Inflammation; Karyotyping; Macrophages; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Muramidase; Plasmacytoma; Rosette Formation

The CSF levels of lactoferrin, lysozyme, and beta 2-microglobulin (beta 2 mu) were measured in patients with evident, probable, or possible inflammatory CNS reactions and compared to those found in neurologically apparently healthy patients. Patients with viral CNS infections had significantly raised beta 2 mu and lysozyme levels but normal lactoferrin levels, indicating a local activation of lymphocytes and monocytes but not of granulocytes. Patients with bacterial CNS infections had significantly raised levels of all three cell markers, but the increase of lysozyme and lactoferrin was relatively more pronounced than that of beta 2 mu, indicating that the inflammatory response to bacterial agents is dominated by monocytes and granulocytes. Patients with primary or secondary malignant brain tumors were characterized by a moderate increase of beta 2 mu and a considerable increase in both lysozyme and lactoferrin, i.e., the same protein pattern as observed in bacterial CNS infection. The lysozyme levels were moderately increased in half the patients with benign cerebral tumors while the levels of beta 2 mu and lactoferrin were normal, indicating that benign and malignant brain tumors induce different local inflammatory CNS reactions. Half the patients with pituitary gland adenoma had elevated beta 2 mu and lysozyme levels but normal lactoferrin levels, suggesting that immunological mechanisms are associated with the adenoma development. Patients with MS had moderately but significantly raised CSF levels of beta 2 mu and lysozyme and a third of them also had raised levels of lactoferrin, a protein pattern suggesting a low-active inflammatory process in CNS involving mononuclears and granulocytes. A similar protein pattern was found in Guillain-Barré syndrome. In cerebrosarcoidosis we noted considerably increased lysozyme and beta 2 mu but normal lactoferrin levels, consistent with the idea that the sarcoid granuloma mass is dominated by monocytic inflammatory cells. The data obtained indicate a clinical value of lactoferrin, lysozyme, and beta 2 mu as differential indices of inflammatory cell reactions taking place in various CNS processes.

Topics: Adult; Albumins; Bacterial Infections; beta 2-Microglobulin; Beta-Globulins; Central Nervous System Diseases; Cerebrospinal Fluid Proteins; Female; Humans; Inflammation; Lactoferrin; Lactoglobulins; Male; Muramidase; Virus Diseases

Topics: Acetylglucosaminidase; Animals; Cells, Cultured; Chronic Disease; Glucuronidase; Hydrolases; Inflammation; Kinetics; Lysosomes; Macrophages; Mice; Muramidase; Peptide Hydrolases; Phagocytosis

The present study was carried out to compare radiological and physiological changes in sarcoidosis with biochemical markers for inflammatory cell populations. Of 53 patients with sarcoidosis, 28 had respiratory symptoms and 30 past or present bilateral hilar adenopathy without symptoms. A clinical score based on lung function tests and radiological findings correlated well with elevations of lysozyme and beta2-microglobulin in serum, indicating increased inflammatory cell activity in patients with more severe lung affection. A covariation between beta2-microglobulin and lysozyme was found, suggesting concomitant activation of macrophages and lymphocytes in sarcoidosis. Serum levels of lactoferrin were elevated in patients with a disease of short duration but did not correlate with the severity of the lung affection. The closing volume also seems to be abnormal in the early course of the disease, while elevated lysozyme and beta2-microglobulin levels rather seem to reflect the extent of the pulmonary affection.

Topics: Adult; Aged; beta 2-Microglobulin; Female; Humans; Inflammation; Lactoferrin; Lung; Lung Diseases; Male; Middle Aged; Muramidase; Radiography; Respiratory Function Tests; Sarcoidosis

The aim of the present study was to characterize leukocytes present in a local inflammatory reaction with respect to production of prostaglandin E (PGE) and the release of factors affecting lymphocyte function, which are produced by macrophages (interleukin-1) or stimulated lymphocytes (interleukin-2). Lewis rats immunized against bovine serum albumin (BSA) were challenged by intraperitoneal injection of antigen. The PGE release by peritoneal cells (PEC) was tested in vitro and found to be enhanced in immunized rats before BSA challenge. However, PEC harvested after the injection of antigen showed a marked reduction in prostaglandin production during the first 24 hr. When these cells were tested for the secretion of lymphokines (IL-1, IL-2) the same depression was found.

Topics: Animals; Arthritis, Experimental; Exudates and Transudates; Inflammation; Interleukin-1; Leukocytes; Muramidase; Prostaglandins E; Proteins; Rats

Comparative evaluation of immunotropic and inflammatory activity of thymus factor (TFX) and levamisole was performed. In numerous experimental systems it was shown that both preparations exert stimulatory effect on the course and intensity of the specific immune reactions. Many tests proved their stimulatory effect on the nonspecifically induced inflammatory processes. TFX as well as levamisole, beside their influence on the course of immune reactions, are also active in the differentiation of T lymphocytes. Differences and similarities in the effect of both evaluated compounds were examined in the in vivo and in vitro systems and it was observed that levamisole possesses the components of activity atypical for thymus hormones.

Topics: Animals; Cathepsins; Graft vs Host Reaction; Guinea Pigs; Hemolysis; Immunity, Cellular; Inflammation; Levamisole; Mice; Mice, Inbred Strains; Muramidase; Phagocytosis; Rabbits; Rats; Rats, Inbred Strains; Thymus Extracts; Tuberculin Test

Propionibacterium acnes cells were tested for the ability to trigger lysosomal hydrolase release from human polymorphonuclear leukocytes. Representative strains of P. acnes serotype I and II failed to stimulate lysosomal release in the absence of serum. P. acnes growth culture supernatants failed to trigger release under any test condition. Addition of fresh or heat-inactivated human serum resulted in lysosomal hydrolase release directly proportional to the number of P. acnes/PMN. Pooled sera from acne patients, with a high anti-P. acnes titer stimulated release to P. acnes. Preabsorption of this reagent with P. acnes cells reduced the anti-P. acnes titer and produced 93.37 +/- 11.49% inhibition of lysosomal enzyme release compared to unabsorbed anti-serum. Electron microscopy indicated that P. acnes was readily phagocytosed by PMNs when fresh or heated serum was present.

Topics: Acne Vulgaris; Humans; In Vitro Techniques; Inflammation; Muramidase; Neutrophils; Phagocytosis; Propionibacterium acnes

Mononuclear phagocytes participate in various stages of chronic inflammatory responses and associated diseases. Such participation is mediated by (a) direct interaction with pericellular interstitial tissue components as well as with other cell types present at sites of inflammation and (b) by secretion of soluble mediators. Several of these mediators are synthesized and secreted in increased amounts after macrophages interact with inflammatory stimuli. In this paper we pay particular attention to neutral proteinases and prostaglandins. It is shown that these 2 classes of mediators are released in significant amounts under different conditions. Prostaglandins are synthesized most readily by resident populations of mouse peritoneal macrophages responding to various model inflammatory stimuli. Mouse peritoneal macrophage populations elicited in vivo by inflammatory stimuli are less responsive in this respect. In contrast neutral proteinase secretion does not occur in resident cell populations but is observed on a continuous basis in elicited populations. Such secretion can be increased further by addition of phagocytic stimuli and initiated in resident populations by model inflammatory stimuli such as phorbol myritate acetate. Other secretory products of macrophages with possible relevance to inflammation are discussed briefly. Finally some of the effects of antiinflammatory glucocorticoids, cyclooxygenase inhibitors and dapsone on the secretory activity of macrophages are briefly summarized.

Topics: Animals; Anti-Inflammatory Agents; Arachidonic Acids; Cell Division; Complement System Proteins; Dapsone; Endopeptidases; Glucocorticoids; Guinea Pigs; Hydrolases; Inflammation; Macrophages; Mice; Muramidase

Host responses to infectious organisms should be modulated so that tissue-damaging products of inflammatory cells do not produce excessive destruction of normal tissue. Lysozyme, which is continuously secreted by monocytes, which, in turn, migrate relatively late to inflammatory areas, was found to significantly dampen several responses of neutrophils to inflammatory stimulants. Thus, human lysozyme obtained and purified from the urine of patients with monocytic leukemia (but not its structurally similar and comparably cationic analogue, eggwhite lysozyme) depresses chemotaxis of normal neutrophils to activated complement, bacterial supernate, and N-formylmethionyl-phenylalanine. In addition, human (but not eggwhite) lysozyme depresses oxidative metabolism (hexose monophosphate shunt activity) and superoxide generation of neutrophils. The specificity of the suppressive effects was indicated by inhibition studies with rabbit antihuman lysozyme antibody, and with the trisaccharide of N-acetylglucosamine, a specific inhibitor of lysozyme. The results suggest that lysozyme, a product of inflammatory cells themselves, may function in a negative feedback system to modulate the inflammatory response.

Topics: Cell Migration Inhibition; Cell Survival; Chemotaxis, Leukocyte; Cyclic AMP; Feedback; Humans; Immune Sera; In Vitro Techniques; Inflammation; Muramidase; Neutrophils; Oxygen Consumption

The concentrations of several polymorphonuclear neutrophilic lysosomal constituents were quantitated by immunochemical and enzymatic assays in 28 inflammatory and 9 noninflammatory synovial fluids. The quantities of lactoferrin, myeloperoxidase, and enzymatically determined lysozyme were covariate with the neutrophil count. Enzymatic activities measured with synthetic substrates developed for the assay of chymotryptic-like cationic protein (cathepsin G) and elastase, along with immunochemically determined lysozyme, were independent of the neutrophil count. Although the latter assays were developed and standardized with human neutrophilic lysosomal constituents, they measure different activities in inflammatory synovial effusions. No elastase was detected if elastin was used as the substrate. Regardless of the source of the enzymes, there was a negative correlation between their concentration and the degree of radiographic destruction of the joint from which the fluid was obtained. Lysosomal enzymes in solution in synovial fluid are not likely to be primarily involved in cartilage destruction.

Topics: Cathepsins; Complement C3; Humans; Inflammation; Lactoferrin; Lysosomes; Muramidase; Neutrophils; Pancreatic Elastase; Peroxidase; Synovial Fluid; X-Rays

Topics: Cell Adhesion; Cell Movement; Cytoplasmic Granules; Exudates and Transudates; Filtration; Humans; Inflammation; Muramidase; Neutrophils; Time Factors

Topics: Animals; Arthritis; Arthritis, Experimental; Complement System Proteins; Dermatitis, Atopic; Diphtheria Antitoxin; Diphtheria Toxin; Hippocampus; Hypersensitivity, Delayed; Inflammation; Muramidase; Rabbits; Rats; Tuberculin Test; Turpentine

Human leukocyte extracts, egg white lysozyme, cationic proteins, polymyxin, colimycin, and phenol are capable of releasing lipoteichoic acids (LTA) from group A streptococci and Strep. mutans. While the extraction of LTA by phenol is optimal at pH 4.7, the release of LTA from streptococci by the other agents is optimal at pH 7.4. LTA released by all agents was found to have the same sensitizing abilities, as determined by passive hemagglutination, and to have a similar chemical composition, as shown by thin-layer chromatography and radioactive scanning. The LTA-releasing capacity of all the agents is strongly inhibited by normal human serum. The possible role played by LTA released by leukocyte factors in the pathogenesis of tissue damage during bacterial infections is discussed.

Topics: Cell Extracts; Chemical Phenomena; Chemistry; Hydrogen-Ion Concentration; Inflammation; Leukocytes; Muramidase; Phosphatidic Acids; Streptococcus; Streptococcus mutans; Streptococcus pyogenes; Teichoic Acids; Tissue Extracts

Topics: Adolescent; Adult; Eye Diseases; Female; Humans; Immunization, Passive; Inflammation; Male; Middle Aged; Muramidase; Tears; Typhoid-Paratyphoid Vaccines

Topics: Animals; Arthritis; Arthritis, Experimental; Blood Platelets; Cell Extracts; Cytarabine; Glucuronidase; Granulocytes; Inflammation; Leukocyte Count; Leukocytes; Muramidase; Mycobacterium; Rats; Time Factors; Tritium; Zymosan

Topics: Antigen-Antibody Complex; B-Lymphocytes; Capillary Permeability; Cell Migration Inhibition; Chemotaxis, Leukocyte; Eosinophils; Granuloma; Humans; Immunologic Deficiency Syndromes; Inflammation; Lymphocyte Activation; Macrophages; Monocytes; Muramidase; NAD; NADP; Necrosis; Neutrophils; Opsonin Proteins; Phagocytes; Phagocytosis; T-Lymphocytes

In order to elucidate the biochemical anti-inflammatory properties of naproxen, the effects of this compound on activities of mucopolysaccharase [beta-glucuronidase (beta-Gase) and lysozyme (LZ)], acid protease (APase) and collagenolytic enzyme (CL) in inflamed tissues were investigated by means of a proliferative inflammatory model in filter-paper-implanted rats. In the preventive test, naproxen strongly inhibited granuloma formation and exudate accumulation as did indomethacin and prednisolone. Although the inhibitory effects of naproxen on all these enzymes were quite evident, indomethacin failed to inhibit APase activity. Prednisolone did not significantly inhibit LZ and APase activities in granuloma. In the curative test, prednisolone caused a marked decrease in the weight of the granuloma already formed and in the volume of the exudate, but with naproxen and indomethacin there was only a slight decrease. Naproxen and indomethacin induced slight but significant inhibition of LZ and CL activities, while prednisolone showing a weak inhibition of CL activity only. From these results, it may be concluded that anti-inflammatory and anti-rheumatic effects of naproxen are partly attributable to its inhibitory actions on these lysosomal enzymes.

Topics: Adenosine Triphosphatases; Animals; Anti-Inflammatory Agents; Exudates and Transudates; Female; Glucuronidase; Glycosaminoglycans; Granuloma; Inflammation; Lyases; Microbial Collagenase; Muramidase; Naphthaleneacetic Acids; Naproxen; Protease Inhibitors; Rats; Time Factors

The studies reviewed in this chapter provide further evidence that the secretion of lysosomal enzymes and other hydrolases is a constitutive function of certain cells whereas in other cells is an inducible process probably contributing to the pathology of a variety of diseases. Little is known of the mechanisms mediating the secretion of lysosomal enzymes. We have summarized evidence suggesting a role of microfilaments and microtubules in controlling enzyme release, but further studies of the biochemical mechanisms which control the activity of these subcellular structures are required. The fusion of lysosomes with the plasma membrane has been observed in several situations and the mechanisms underlying processes of this nature have been studied in lower organisms (Satir et al. 1973; Plattner 1974). Agents, such as concanavalin A, which interfere with the fusion of endosomes with lysosomes (Goldman 1974; Edelson and Cohn 1974a, b) should also be useful in determining the chemical nature of membrane components involved in the fusion process. New information on the fate of secreted acid hydrolases has been obtained from studies of the uptake of lysosomal enzymes by fibroblasts. Clearly, the mechanisms by which these cells endocytose secreted lysosomal enzymes will be a subject for detailed study in view of the important of directing enzymes and drugs into lysosomes (De Duve et al. 1974). The mechanisms by which extracellular inhibitors inactivate hydrolytic enzymes, particularly proteinases, is also being clarified (for review see Davies 1975) and this should aid in finding new ways for preventing tissue damage caused by the excessive secretion of these enzymes. Further investigation concerning the secretion of lysosomal enzymes should establish the essential physiological role which these enzymes play at both extracellular and intracellular sites. Also, a close examination of the interaction of both endogenous and exogenous stimuli of inflammation with cells resulting in the secretion of hydrolytic enzymes, will clarify the mechanisms underlying the initiation and progression of the inflammatory process in its diverse forms.

Topics: Animals; Anti-Inflammatory Agents; Antigen-Antibody Complex; Bone Resorption; Carrageenan; Cell Transformation, Neoplastic; Cell Wall; Complement System Proteins; Cytochalasin B; Dental Plaque; Glucuronidase; Humans; Hydrolases; Inflammation; Lysosomes; Macrophages; Microbial Collagenase; Muramidase; Neutrophils; Nucleotides, Cyclic; Plasminogen Activators; Pneumoconiosis; Streptococcus pyogenes

The effect of tetracycline and oleandomycin on the complement titer, lysozyme content, serum bactericidal properties and presence of specific antibiotic antibodies in the blood serum was studied. The latter were shown with the Hoigné reaction under conditions of aseptic inflammation caused against the background of latent tetanus intoxication. It was shown that tetracycline and oleandomycin used in treatment of the animals with aseptic inflammation developed at the background of latent tatanus intoxication induced an increased in the complement titer, lysozyme content and bactericidal properties of the serum. Reduction of the above indices was observed by the 15th-20th day after discontinuation of the drug use. The increase in the factors of non-specific immunity under the effect of tetracycline and oleandomycin in the animals with aseptic inflammation caused against the background of latent tetanus intoxication was accompanied by appearance in the blood serum on non-specific antibodies revealed with the Hoigné reaction. Changed reactivity because of latent tetanus intoxication was accompanied by a delay in the formation of the non-specific antibodies in the blood serum. However, later the rate of their accumulation became higher and as a result the maximum titers of the antibodies were 2-3 times higher than those in the control animals.

Topics: Animals; Antibody Specificity; Antigen-Antibody Reactions; Blood Bactericidal Activity; Complement Fixation Tests; Immunity; Inflammation; Muramidase; Oleandomycin; Rabbits; Tetanus; Tetracycline; Time Factors

Topics: Alkaline Phosphatase; Angiogenesis Inducing Agents; Animals; Catalase; Delayed-Action Preparations; Inflammation; Molecular Weight; Muramidase; Polymers; Rabbits; Structure-Activity Relationship; Trypsin Inhibitors

A new methodological approach to the study of the organism resistance to infection with the use of experimental animals with controlled microflora (germfree and other categories of gnotobiotic animals) is considered. Characteristics of the state of natural resistance of germfree animals, revealing considerable defects of cellular and humoral protection mechanisms, are given. Findings of experimental studies into inflammation, phagocytosis and other reactions of nonspecific resistance in gnotobiotic animals, disclosing complex mechanisms of formation of these reactions under the influence of microflora, are presented. The etiological and pathogenetic role of the microbial factor in the development of infectious diseases and in the formation of mechanisms of protective reactions at various levels of integration of the organism are discussed. Conclusions concerning prospects of the gnotobiological approach in investigating the role of the microbic factor in pathology are set forth.

Topics: Animals; Antibody Formation; Bacterial Infections; Burns; Cell Movement; Complement System Proteins; Germ-Free Life; Growth; Guinea Pigs; Immunity, Cellular; Infections; Inflammation; Intestines; Leukocytes; Mice; Muramidase; Phagocytosis; Properdin; Rats; Virus Diseases

This review deals with those biological activities of peptidoglycan that are not directly analogous to the properties of gram-negative bacterial endotoxin. The report is divided into 3 major parts: 1. A survey of peptidoglycan activities such as the induction of inflammatory skin reactions, lesion-enhancing activity (virulence factor), inhibition of phagocytosis of bacteria, inhibition of cell migration, cytotoxicity to mammalian cells, potentiation of the humoral and cellular immune response (adjuvant activity) and enhancement of tumor defense in experimental animals. 2. A presentation of factors which may influence these biological activities of peptidoglycan. 3. A brief discussion of the potential mechanisms of action of peptidoglycan.

Topics: Adjuvants, Immunologic; Animals; Antibodies, Bacterial; Bacillus megaterium; Cell Migration Inhibition; Chemotaxis; Inflammation; Muramidase; Necrosis; Neutralization Tests; Peptidoglycan; Phagocytosis; Skin Manifestations; Staphylococcal Infections; Staphylococcus; Streptococcus; Virulence

Topics: Adjuvants, Immunologic; Bacteria; Cell Wall; Inflammation; Muramidase; Peptidoglycan

Topics: Animals; Anti-Inflammatory Agents; Glucuronidase; Inflammation; Microbial Collagenase; Muramidase; Naphthalenes; Peptide Hydrolases; Propionates; Rats

Topics: Animals; Antigen-Antibody Complex; Arthritis, Rheumatoid; Complement System Proteins; Female; Guinea Pigs; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Inflammation; Knee Joint; Leukocyte Count; Male; Middle Aged; Muramidase; Neutrophils; Rheumatoid Factor; Solubility; Synovial Fluid

Topics: Aminopyrine; Animals; Anti-Inflammatory Agents; Aspirin; Dexamethasone; Enzyme Activation; Female; Flufenamic Acid; Glucuronidase; Glycoside Hydrolases; Hexosaminidases; Indomethacin; Inflammation; Muramidase; Phenylbutazone; Prednisolone; Rats; Time Factors

Topics: Animals; Arthritis; Arthus Reaction; Cell Adhesion; Cell Movement; Complement System Proteins; Connective Tissue; Cytoplasmic Granules; Glucuronidase; Humans; In Vitro Techniques; Inflammation; Microscopy, Electron; Muramidase; Nephritis; Neutrophils; Peroxidases; Phagocytosis; Rabbits

Topics: Adult; Blood; Cathepsins; Exudates and Transudates; Female; Humans; Immunity, Cellular; Inflammation; Leukocytes; Male; Middle Aged; Muramidase; Phagocytosis; Skin Window Technique; Staphylococcus

Topics: Adrenocorticotropic Hormone; Aminobenzoates; Anti-Inflammatory Agents; Dental Pulp Capping; Edema; Enzyme Therapy; Female; Glucocorticoids; Humans; Indomethacin; Inflammation; Male; Mouth Diseases; Muramidase; Nicotinic Acids; Oxyphenbutazone; Postoperative Complications; Pyrazoles; Salicylates; Stomatitis

Topics: Blood Platelets; Clinical Enzyme Tests; Erythrocytes; Exudates and Transudates; Hemolysis; Humans; Inflammation; Leukocytes; Lymphocytes; Monocytes; Muramidase; Skin Window Technique; Staphylococcal Infections

Topics: Animals; Chlorides; Chymotrypsin; Drug Synergism; Dry Ice; Guinea Pigs; Histamine; Inflammation; Injections, Intramuscular; Injections, Intraperitoneal; Lithium; Muramidase; Prednisolone; Rats; Salicylates

Topics: Adenoids; Adolescent; Anabolic Agents; Asthma; Child; Child, Preschool; Chronic Disease; Humans; Immune Sera; Inflammation; Muramidase; Pneumonia; Saliva; Skin Tests; Tonsillitis

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Cell Biology; Cell Movement; Chemotaxis; Culture Techniques; Glucuronidase; Inflammation; Leukocytes; Lysosomes; Macrophages; Muramidase; Neutrophils; Rabbits

Topics: Administration, Oral; Anti-Infective Agents; Bacterial Infections; Case-Control Studies; Female; Histiocytes; Humans; Inflammation; Male; Muramidase; Phagocytosis; Skin Window Technique; Wound Healing

Topics: Animals; Antibodies; Body Fluids; Extracellular Space; Exudates and Transudates; Inflammation; Muramidase; Rabbits

A chronic infiltration of polymorphonuclear leukocytes was invariably found in the infertile regions of uteri containing foreign bodies in conventional rats, germfree rats, mice, and rabbits. Polymorphonuclear leukocytes were never found in the fertile regions of these uteri. A foreign body in the uterus of the rat, and probably also the mouse, was associated with a bacterial infection which spread the inflammatory response throughout the horn containing the foreign body, and in the mouse occasionally into the control horn as well. No bacteria could be cultured from the rabbit uterine horn containing a foreign body. In the germfree rat, both the infiltration of polymorphonuclear leukocytes into the uterus and fertility were significantly different from that observed in the conventional rat. Whereas in the conventional rat the inflammation and infertility extended along the entire length of the uterine horn containing a small foreign body, in the germfree rat the inflammation and infertility were closely correlated to the position of the foreign body. As judged by measurements of lysozyme in the uterine lumens of rats and rabbits, polymorphonuclear leukocytes released their contents into solution in the uterine lumen. It is concluded that some substance derived from polymorphonuclear leukocytes may exert toxic effects on fertilized ova or on spermatozoa and thus be responsible for the infertility of uteri containing foreign bodies.

Topics: Animals; Culture Techniques; Female; Foreign Bodies; Germ-Free Life; Infertility, Female; Inflammation; Intrauterine Devices; Mice; Muramidase; Neutrophils; Rabbits; Rats; Uterus

The roles of uterine inflammation and infection in creating an environment hostile to fertilized eggs or spermatozoa which would explain the contraceptive action of IUDs were investigated in the rat, rabbit, and mouse. Inflammation, as evidenced by the appearance of polymorphonuclear leucocytes, was always present along with a foreign body, and inflamed areas corresponded to regions of known infertility: the entire length of the horn containing the foreign body in the rat uterus, the corresponding horn as well as part of the control horn in the mouse, and only the tissue in contact with the foreign body in the rabbit. Acute inflammatory response (polymorphonuclear stage) persisted indefinitely after insertion of the IUD. Lysozyme measurements in rat and rabbit uteri indicated that polymorphonuclear leucocytes released their contents into the lumen in infertile regions. Cultured rat uteri containing foreign bodies were found to contain over 100 million bacteria of mixed species (compared to none in controls), while rabbit uteri appeared bacteria free. The effects of inflammation were separable from those of infection, however, with the use of germ-free rats. The presence of a foreign body in these rats was associated with a very localized inflammation, with lysozyme only in the segment in contact with the foreign body. Fertilized eggs entered and implanted in the noninflamed region when the foreign body was in the cervical end of the horn; when it was in the ovarian end, all ova were killed, indicating that the antifertility effect of the IUD was possible in absence of bacteria. It is concluded that some component of the polymorphonuclear leucocyte may kill fertilized eggs and thus be responsible for the infertility of uteri containing IUDs.

Topics: Animals; Female; Germ-Free Life; Inflammation; Intrauterine Devices; Leukocytes; Mice; Muramidase; Pregnancy; Rabbits; Rats; Uterine Diseases; Uterus

Topics: Adrenalectomy; Animals; Bradykinin; Chymotrypsin; Deoxyribonucleases; Edema; Fibrinolysin; Granuloma; Inflammation; Male; Muramidase; Pancreatin; Peptide Hydrolases; Rats; Serratia; Trypsin

Lysosomal granules of rabbit exudate polymorphonuclear (PMN) leucocytes were isolated and then lysed by freezing-thawing. Topical application of this material to rat and rabbit mesentery produced sticking and emigration of leucocytes, stasis of blood flow, and petechial hemorrhage. The granule-free, supernatant fraction of the homogenized leucocytes failed to produce any of these reactions. Cationic proteins extracted from these granules by weak acid and precipitated by ethanol at concentrations of 20 and 45 per cent, were also tested on heterologous, homologous, and autologous mesenteric vessels. The 20 per cent ethanol-precipitated fraction produced all of the aforementioned injury reactions, whereas the 45 per cent fraction was inactive. The intensity of inflammatory changes produced by the active cationic protein fraction was greater than that produced by lysed whole granules. Both the 20 per cent and 45 per cent ethanol fractions of cationic protein induced clumping of rabbit platelets, in vitro. The 20 per cent ethanol fraction also caused a slight acceleration in rate of swelling of isolated rabbit liver mitochondria. The active material proved to be non-pyrogenic in rabbits. This material exhibited no kinin-like effects when tested on isolated smooth muscle preparations (rabbit aorta and guinea pig ileum). In the rat, the protein produced a transient vasodepression which was inhibited by pretreatment of the animal with an antihistamine. Ultraviolet absorption data and ribose assays showed that the 20 per cent ethanol fraction contained only 4 per cent or less of ribonucleic acid. Upon electrophoresis in starch gel, using acid buffer, this fraction separated into at least three major components which migrated towards the cathode. Precipitation of one of the slowly migrating components by titration of the fraction to pH 10.5 greatly increased the inflammatory activity of the material. The inflammatory basic protein fraction was essentially devoid of acid phosphatase, beta glucuronidase, acid ribonuclease, lysozyme, and catalase activity. The non-inflammatory basic protein fraction contained appreciable quantities of acid ribonuclease and lysozyme. The foregoing data demonstrate that certain of the cationic proteins present in lysosomes of rabbit exudate PMN leucocytes can reproduce one of the cardinal features of the inflammatory response; namely, adhesion and emigration of leucocytes in the microcirculation. These findings offer fresh support for t

Topics: Acid Phosphatase; Animals; Bradykinin; Capillary Permeability; Cytoplasmic Granules; Exudates and Transudates; Glucuronidase; Guinea Pigs; Histamine; Inflammation; Leukocytes; Lysosomes; Mesentery; Microcirculation; Mitochondria; Muramidase; Muscle, Smooth; Neutrophils; Norepinephrine; Pharmacology; Promethazine; Proteins; Rabbits; Rats; Research; Ribonucleases; RNA; Toxicology

Topics: Anti-Infective Agents, Local; Antiviral Agents; C-Reactive Protein; Dermatologic Agents; Disease; Eubacterium; Female; Humans; Inflammation; Muramidase; Pelvic Inflammatory Disease; Pelvis

Topics: Anti-Infective Agents, Local; Dermatologic Agents; Humans; Inflammation; Muramidase; Palatine Tonsil; Tonsillitis

Topics: Anti-Infective Agents, Local; Blood; Dermatologic Agents; Humans; Inflammation; Muramidase; Palatine Tonsil; Tonsillitis