interleukin-8 and Leukocyte-Adhesion-Deficiency-Syndrome

Topics: Apoptosis; Chediak-Higashi Syndrome; Familial Mediterranean Fever; Humans; Inflammation; Interleukin-1; Interleukin-8; Leukocyte-Adhesion Deficiency Syndrome; Neutropenia; Neutrophil Activation; Neutrophils; Phagocytosis

The role of neutrophils in the pathogenesis of Salmonella enterica Typhimurium-induced ruminant and human enteritis and diarrhea has yet to be characterized with in vivo models. To address this question, the in vivo bovine ligated ileal loop model of nontyphoidal salmonellosis was used in calves with the naturally occurring bovine leukocyte adhesion deficiency (BLAD) mutation whose neutrophils are unable to extravasate and infiltrate the extravascular matrix. Data obtained from 4 BLAD Holstein calves homozygous for BLAD (CD18-), 1 to 5 weeks of age, were compared with 4 controls, age-matched Holstein calves negative for BLAD (CD18+). Morphologic studies revealed that infection of CD18- calves with S Typhimurium resulted in no significant tissue infiltration by neutrophils, less tissue damage, reduced luminal fluid accumulation, and increased bacterial invasion, when compared with CD18+ calves. Ultrastructurally, lesions in enterocytes induced by S Typhimurium infection in CD18- calves--including attachment and disruption of the brush border, apical membrane ruffling formation, and cellular degeneration--were similar to the ones reported in the literature for CD18- calves. Study of cytokine gene expression by quantitative real-time polymerase chain reaction revealed that early stages of acute infection (4-8 hours postinfection) were associated with increased interleukin 8 gene expression in the absence of tissue influx of neutrophils in CD18- calves, whereas later stages of infection (12 hours postinfection) were associated with increased expression of growth-related oncogene alpha in the presence of neutrophil influx in CD18+ calves. In contrast, the proinflammatory cytokines interleukin 1beta and tumor necrosis factor alpha were poorly correlated with the presence or absence of tissue neutrophils.

Topics: Animals; Animals, Suckling; Cattle; Cattle Diseases; CD18 Antigens; Chemokine CXCL1; Female; Histocytochemistry; In Vitro Techniques; Interleukin-1beta; Interleukin-8; Leukocyte-Adhesion Deficiency Syndrome; Male; Microscopy, Electron, Scanning; Microscopy, Electron, Transmission; Peyer's Patches; Reverse Transcriptase Polymerase Chain Reaction; RNA; Salmonella Infections, Animal; Salmonella typhimurium; Tumor Necrosis Factor-alpha

Serum cytokine levels and their expression of mRNA on neutrophils from a bone marrow (BM) transplanted heifer with leukocyte adhesion deficiency were evaluated. The clinical condition of the affected heifer was relatively stable after BM-transplantation. Persistent hyper gamma-globulinemia and increased serum immunoglobulin G (IgG) concentrations were monitored longitudinally. The concentration of interleukin (IL)-1beta in serum from the affected heifer ranged from 15.8 to 321.7 ng/ml, and maximum concentration occurred at the time which coincided with peak IL-6. Serum levels of IL-6 ranged from 0.32 to 27.9 ng/m l, and they appeared to be associated with the increment of serum IgG in the affected heifer. mRNAs for IL-1beta, IL-6, IL-8 and granulocyte and macrophage colony stimulating factor (GM-CSF) were increased in neutrophils from the affected heifer compared to controls. Persistent hyper gamma-globulinemia of the affected heifer appeared to be associated with enhanced mRNA expression for IL-6 and its serum levels. These findings suggest that humoral immunity of the affected heifer is activated and the production of neutrophils appears to be enhanced under the incapability of beta(2) integrin-mediated functions of phagocytic cells.

Topics: Animals; Bone Marrow Transplantation; Cattle; Cattle Diseases; Cytokines; Female; Gene Expression; Granulocyte-Macrophage Colony-Stimulating Factor; Hypergammaglobulinemia; Immunoglobulin G; Interleukin-1; Interleukin-6; Interleukin-8; Leukocyte-Adhesion Deficiency Syndrome; Neutrophils; RNA, Messenger

Changes in interleukin (IL)-1beta, IL-6 and IL-8 in serum, and their mRNA expression on neutrophils from a 4.6-month old Holstein young calf with bovine leukocyte adhesion deficiency (BLAD) during the acute phase were evaluated. IL-1beta concentrations in the serum of the calf with BLAD at age 143-162 days ranged from 8.7 to 16.6 ng/ml, whereas the values were less than 2.7 ng/ml in control calves. Serum IL-6 (0.04 ng/ml) was only detected on the 1st day when the animal was diagnosed with the BLAD. IL-1beta and IL-8 mRNA expression on neutrophils from the affected calf appeared to be similar to those of controls. Serum cytokine levels and their mRNA expression on neutrophils from the calf with BLAD appeared to be little affected by the deficient expression of beta(2)-integrin on leukocytes, and are considered to be modulated by the inflammatory stimuli.

Topics: Acute-Phase Reaction; Animals; Cattle; Cattle Diseases; Gene Expression Regulation; Genetic Diseases, Inborn; Interleukin-1; Interleukin-6; Interleukin-8; Leukocyte-Adhesion Deficiency Syndrome; Neutrophils; RNA, Messenger

Human exudative neutrophils have greatly increased stores of the neutrophil chemoattractant IL-8 compared with peripheral blood cells, but the mechanism for the increase is not defined. In this report, we show that treatment of peripheral blood neutrophils with the chemotactic peptide fMLP or with leukotriene B(4) or fibrinogen results in little increase in the production of IL-8 by peripheral blood neutrophils. However, a chemotactically active dose of fMLP (5 x 10(-9) M) or leukotriene B(4) (1 x 10(-7) M) in the presence of a physiological concentration (2 mg/ml) of fibrinogen results in a receptor-mediated, pertussis toxin-sensitive, synergistic 30-fold increase in IL-8 synthesis. The levels of IL-8 attained are comparable to those observed in exudative cells. Higher concentrations of fMLP (1 x 10(-7) M) are associated with reduced IL-8 protein synthesis without IL-8 degradation, indicating a sensitive regulatory mechanism for IL-8 production. Treatment of neutrophils with fibrinogen and fMLP resulted in minimal changes in the steady state levels of mRNA for macrophage inflammatory protein-1alpha and -1beta and monocyte chemoattractant protein-1. In contrast, in the presence of fibrinogen, the steady-state level of neutrophil IL-8 mRNA increased 8-fold with 5 x 10(-9) M fMLP but was not decreased with 1 x 10(-7) M fMLP, suggesting that neutrophils are specifically adapted to modulate neutrophil IL-8 synthesis through transcriptional and posttranscriptional mechanisms. The data indicate that fibrinogen can function not only as a substrate in the clotting cascade, but also as an important effector during the evolution of the innate immune response.

Topics: Calcium; Chemokine CCL2; Chemokine CCL4; Cycloheximide; Dose-Response Relationship, Drug; Drug Synergism; Fibrinogen; Humans; In Vitro Techniques; Interleukin-8; Leukocyte-Adhesion Deficiency Syndrome; Leukotriene B4; Macrophage Inflammatory Proteins; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Pertussis Toxin; Protein Synthesis Inhibitors; Receptors, Formyl Peptide; Receptors, Immunologic; Receptors, Leukotriene B4; Receptors, Peptide; RNA, Messenger; Virulence Factors, Bordetella

Neutrophils isolated from a child with severe leukocyte adhesion deficiency 1 (LAD1) had a complete absence of expression of the CD11/CD18 beta2 integrin family of adhesion molecules, and were shown to be deficient in the in vitro adhesion and migration properties. However, we found that interleukin-8 (IL8), a potent chemoattractant for neutrophils, and sputum sol phase induced these LAD1 neutrophils to migrate through an endothelial cell layer in vitro, and confirmed that this migration was CD18-independent. These findings add to evidence of CD18-independent mechanisms of neutrophil recruitment, in particular neutrophil infiltration into the lungs, where IL8 may be an important recruitment factor.

Topics: Antibodies; CD11 Antigens; CD18 Antigens; Cell Adhesion; Cell Movement; Cells, Cultured; Diffusion Chambers, Culture; Endothelium, Vascular; Flow Cytometry; Humans; Infant; Integrin alphaXbeta2; Interleukin-8; Leukocyte-Adhesion Deficiency Syndrome; Lymphocyte Function-Associated Antigen-1; Macrophage-1 Antigen; Male; Membrane Glycoproteins; Neutrophil Infiltration; Neutrophils; Precipitin Tests

Platelets bound to thrombogenic surfaces have been shown to support activation-dependent firm adhesion of neutrophils in flow following selectin-mediated tethering and rolling. The specific receptor(s) responsible for mediating adhesion-strengthening interactions between neutrophils and platelets has not previously been identified. Furthermore, the ability of adherent platelets to support the migration of bound neutrophils has not been tested. We studied neutrophil interactions with activated, surface-adherent platelets as a model for leukocyte binding in vascular shear flow and emigration at thrombogenic sites. Our results demonstrate that the beta 2-integrin Mac-1 (CD11b/CD18) is required for both firm attachment to and transmigration of neutrophils across surface-adherent platelets. In flow assays, neutrophils from patients with leukocyte adhesion deficiency-1 (LAD-I), which lack beta 2-integrin receptors, formed P-selectin-mediated rolling interactions, but were unable to develop firm adhesion to activated platelets, in contrast to healthy neutrophils, which developed firm adhesion within 5 to 30 seconds after initiation of rolling. Furthermore, the adhesion-strengthening interaction observed for healthy neutrophils could be specifically inhibited by monoclonal antibodies (mAbs) to Mac-1, but not to lymphocyte function-associated antigen-1 (LFA-1; CD11a/CD18) or intercellular adhesion molecule-2 (ICAM-2; CD102). Further evidence for a beta 2-integrin-dependent neutrophil/platelet interaction is demonstrated by the complete inhibition of interleukin (IL)-8-induced neutrophil transmigration across platelets bound to fibronectin-coated polycarbonate filters by mAbs to Mac-1. Thus, Mac-1 is required for firm adhesion of neutrophils to activated, adherent platelets and may play an important role in promoting neutrophil accumulation on and migration across platelets deposited at sites of vascular injury.

Topics: Blood Platelets; CD18 Antigens; Cell Adhesion; Cell Movement; Humans; Interleukin-8; Leukocyte-Adhesion Deficiency Syndrome; Macrophage-1 Antigen; Neutrophils; P-Selectin