lactoferrin and Hypersensitivity

Lactoferrin is a multifunctional member of the transferrin family of nonheme iron-binding glycoproteins. Lactoferrin is found at the mucosal surface where it functions as a prominent component of the first line of host defense against infection and inflammation. The protein is also an abundant component of the specific granules of neutrophils and can be released into the serum upon neutrophil degranulation. While the iron-binding properties were originally believed to be solely responsible for the host defense properties ascribed to lactoferrin, it is now known that other mechanisms contribute to the broad spectrum anti-infective and anti-inflammatory roles of this protein. In this article, current information on the functions and mechanism of action of lactoferrin are reviewed, with particular emphasis on the activities that contribute to this protein's role in host defense. In addition, studies demonstrating that lactoferrin inhibits allergen-induced skin inflammation in both mice and humans, most likely secondary to TNF-alpha (tumor necrosis factor alpha) production, are summarized. Collectively, these results suggest that lactoferrin functions as a key component of mammalian host defense at the mucosal surface.

Topics: Animals; Anti-Inflammatory Agents; Host-Parasite Interactions; Humans; Hypersensitivity; Immunity, Innate; Immunity, Mucosal; Inflammation; Lactoferrin

The clinical relevance of immunoglobulin E (IgE) to plant glycans is a longstanding debate. We sought to evaluate their clinical reactivity using the human glycoprotein lactoferrin expressed in rice.. Allergic patients with IgE antibodies against plant glycans were analyzed for the presence of IgE against rice-produced lactoferrin. The potency of IgE to induce mediator release was assessed by basophil histamine release and skin prick tests (SPTs). Clinical relevance was evaluated by double-blind placebo-controlled oral challenge (DBPCOC).. Twenty-four of 29 sera (82.7%) with IgE antibodies against plant glycans demonstrated IgE binding to transgenic lactoferrin. In three of five cases transgenic lactoferrin induced histamine release. Compared to a control major grass pollen allergen lactoferrin concentrations needed for biological activity of IgE were 5-6 orders of magnitude higher. Skin prick test and DBPCOC were negative in five patients with potential clinical reactivity that volunteered to undergo these in vivo challenges.. Poor or no biological activity and lack of clinical relevance of IgE-binding plant glycans (five out of five) was demonstrated using human lactoferrin expressed in rice as a model.

Topics: Adolescent; Allergens; Basophil Degranulation Test; Carrier Proteins; Child; Double-Blind Method; Female; Histamine Release; Humans; Hypersensitivity; Immunoglobulin E; Lactoferrin; Male; Middle Aged; Oryza; Phleum; Plant Proteins; Plants, Genetically Modified; Pollen; Polysaccharides; Radioallergosorbent Test; Recombinant Proteins; Skin Tests

Atopic individuals have previously been shown to have an autonomic imbalance consisting of heightened cholinergic responsiveness in the lung, skin, and eyes, and beta-adrenergic hyporesponsiveness in the lungs, eyes, and vasculature. This array of abnormalities is often accompanied by nonspecific bronchial hyperresponsiveness, as well as alpha-adrenergic hyperresponsiveness in individuals with asthma.. To determine whether atopic individuals have intrinsic nasal airway hyperresponsiveness to methacholine, 21 nonatopic subjects and 37 subjects with allergic rhinitis were studied. All subjects were studied out of their allergy seasons, and all allergy-related medications were discontinued before the study began. Subjects underwent nasal challenge with methacholine (1 to 25 mg), and lavaged nasal secretions were analyzed for total protein, the plasma marker albumin, and the glandular marker lactoferrin.. Atopic subjects demonstrated increased glandular responsiveness to methacholine as evidenced by an increase in the secretion of lactoferrin in response to individual doses of methacholine. Although the maximal lactoferrin secretion did not increase, glandular sensitivity to methacholine was heightened because the dose of methacholine required to induce lactoferrin secretion achievable by 60% of the study population was significantly lower in the atopic group. The volume of lavaged secretions recovered and congestion scores were also higher in the atopic group as compared with the normal control group.. These data strongly suggest that atopic individuals have intrinsic nasal glandular hyperresponsiveness to cholinergic stimulation.

Topics: Adolescent; Adult; Dose-Response Relationship, Drug; Female; Humans; Hypersensitivity; Lactoferrin; Male; Methacholine Chloride; Middle Aged; Nasal Cavity; Nasal Mucosa; Nasal Provocation Tests; Proteins; Seasons; Serum Albumin

Both up- and down-regulation of the Toll-like receptors (TLRs) and antimicrobial peptides (AMPs) of the sinonasal mucosa have already been associated with the pathogenesis of chronic rhinosinusitis with (CRSwNP) or without (CRSsNP) nasal polyps. The objective of this study was to determine the expression of all known TLR and several AMP genes and some selected proteins in association with allergy, asthma and aspirin intolerance (ASA) in CRS subgroups. RT-PCR was applied to measure the mRNA expressions of 10 TLRs, four defensins, lysozyme, cathelicidin and lactoferrin (LTF) in sinonasal samples from patients with CRSsNP (n = 19), CRSwNP [ASA(-): 17; ASA(+): 7] and in control subjects (n = 12). Protein expressions were detected with immunohistochemistry (n = 10). Statistical analysis was done with the Kruskal-Wallis ANOVA, Mann-Whitney U, and Student t test. TLR2, TLR5, TLR6, TLR7, TLR8, TLR9, β-defensins 1 and 4, cathelicidin and LTF mRNA expressions were significantly (p < 0.05) increased in CRSwNP, whereas only TLR2 and LTF were up-regulated in CRSsNP compared to controls. There was no statistical difference in respect of allergy, aspirin intolerance and smoking between CRSsNP, ASA(-) and ASA(+) CRSwNP patients. TLR2, TLR3, TLR4, LTF, β defensin 2 and lysozyme protein expressions were found to be elevated in macrophages of CRSwNP samples (p < 0.05). Gene expression analysis showed markedly different expressions in CRSwNP (6 out of 10 TLR and 4 out of 7 AMP genes were up-regulated) compared to CRSsNP (1/10, 1/7). The distinct activation of the innate immunity may support the concept that CRSsNP and CRSwNP are different subtypes of CRS. These findings were found to be independent from allergy, asthma, smoking, aspirin intolerance and systemic steroid application.

Topics: Adult; Antimicrobial Cationic Peptides; beta-Defensins; Case-Control Studies; Cathelicidins; Chronic Disease; Female; Humans; Hypersensitivity; Lactoferrin; Male; Middle Aged; Nasal Polyps; Rhinitis; RNA, Messenger; Sinusitis; Toll-Like Receptors; Young Adult

A highly sensitive method based on mass-barcoded gold nanoparticles (AuNPs) and immunomagnetic separation has been developed for multiplex allergy diagnosis by MALDI mass spectrometry in a component-resolved manner. Different analytical probes were prepared by coating AuNPs with individual allergenic proteins and mass barcode, represented by polyethylene glycol molecules of various chain lengths. Magnetic beads (MBs) functionalized with antihuman IgE antibodies (Abs) were used as immunomagnetic capture probes. IgE Abs were extracted from a patient's blood serum by the formation of a sandwich structure between the AuNPs and MBs. Multiple specific IgE Abs were simultaneously identified by mass spectrometry detection of the mass barcodes, providing an efficient component-resolved allergy diagnosis. Because of the signal amplification provided by the mass barcodes, the developed diagnosis method is very sensitive, with a limit of detection down to picograms per milliliter level for specific IgE Abs. The method can be potentially useful when the sample amount is highly limited and a multiplex diagnostic procedure is required.

Topics: Animals; Cattle; Gold; Humans; Hypersensitivity; Immunoglobulin E; Immunomagnetic Separation; Lactoferrin; Metal Nanoparticles; Milk; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Lactoferrin (LF), a glycogen of the transferrin family with anti-bacterial and immunomodulatory properties, is expressed in various secretions and tissues. Cutaneous LF serves as a mast cell stabilising compound, modulates T cell activity and is found during IgE-mediated late phase reactions at allergen challenged sites. Culicoides hypersensitivity (CHS) in horses is a common IgE-mediated allergic dermatitis, characterised by an early and late phase cutaneous reaction upon allergen challenge. The aim of the study presented here was to examine whether LF mRNA expression in skin biopsies from horses affected by CHS prior to and 4h following intradermal challenge with a commercial C. nubeculosus extract is modified in comparison to skin biopsies from non-affected horses. In order to obtain reliable data, real time PCR was performed and genes of interest were normalized using three different housekeeping genes, beta-actin, GAPDH, beta-2-microglobulin. In comparison to non-affected horses, higher variation in LF mRNA levels both prior to and post-intradermal challenge with C. nubeculosus extract was seen in horses affected by CHS. However, the statistical analysis demonstrated that LF mRNA expression was not significantly different between CHS affected and non-affected horses prior to intradermal challenge with C. nubeculosus extract. Intradermal injection of C. nubeculosus extract did not result in local upregulation of LF mRNA at 4h post-injection. LF mRNA expression was therefore not significantly different pre- or post-intradermal challenge with C. nubeculosus extract in either group. Our data indicate that clinically normal skin of horses affected by CHS is not characterized by modified maintenance levels of LF mRNA. In contrast to human skin allergen challenged sites, LF mRNA levels in horses affected by CHS are not significantly different to that of control sites at 4h post-injection of C. nubeculosus extract.

Topics: Animals; Ceratopogonidae; Horse Diseases; Horses; Hypersensitivity; Insect Bites and Stings; Lactoferrin; Mast Cells; Skin Diseases

Lactoferrin (LF) is an iron-binding glycoprotein present in the cytoplasmic granules of neutrophils and in external secretions of mammals. Although the biological role of human and bovine lactoferrin has been extensively studied, there is still uncertainty as to the nature and function of lactoferrin receptors. We recently determined that methyl-alpha-D-mannopyranoside given intraperitoneally (i.p.) could suppress the adjuvant activity of LF in the generation of delayed-type hypersensitivity (DTH) to ovalbumin (OVA). We concluded that the lactoferrin effects in DTH are mediated by carbohydrate-recognizing receptors like the mannose receptor (MR). This study indicates that subcutaneous (s.c.) administration of very small doses of the Man-bovine serum albumin (Man-BSA) complex, together with a sensitizing dose of the antigen, gives the same effects as i.p. administration of methyl-alpha-D-mannopyranoside. The latter is also a blocker of MR, although of a much lower affinity to the receptor than Man-BSA. The blocking of the adjuvant effect of LF by the Man-BSA complex (when given together with the sensitising dose of antigen) suggests that the function of antigen-presenting cells in the skin (presumably immature dendritic cells expressing MR) is inhibited. The results of our study indicate that a receptor with an affinity for mannose is essential for the mediation of adjuvant lactoferrin function in the generation of DTH.

Topics: Adjuvants, Immunologic; Animals; Female; Hypersensitivity; Lactoferrin; Male; Mannose; Mice; Mice, Inbred CBA; Ovalbumin; Serum Albumin, Bovine

Phagocytic number (PN) and phagocytic index (PI) of neutrophils isolated from blood of patients with autoimmune diseases, allergy to Staphylococcus aureus and from blood of healthy individuals were examined. Our results concerning the influence of lactoferrin (Lf); (6.7 mg/l) on the PI of PMN showed that: 1) Lf enhances reliable PI of PMN at the 30-th minute starting the phagocytic reaction in patients with autoimmune disease in an active stage, in blood donors treated as healthy with the presence of autoantibodies, in patients with autoimmune diseases and proved autoantibodies against tissue, cell antigens and collagen, 2) Lf influences non-significantly PI of PMN in patients with autoimmune collagen diseases in remission, 3) Lf increases PI of PMN with 19% only in 58% from the assessed patients with Staphylococcus aureus, and 4) Lf decreases non-significantly PI of PMN in the healthy controls. Our studies on the effect of Lf on the phagocytic activity of PMN suggest that Lf has stronger effect on the PN compared to the PI: 1) Lf enhances with 86% the PN in patients with Staphylococcus aureus, 2) Lf increases PN of PMN in all of the assessed patients with autoimmune collagen diseases in active stage (mean with 72%), and 3) Lf increases PN of PMN in 4 from the 5 investigated healthy controls (mean with 22%). Our results show a "corrective" effect of Lf on the phagocytic functions in the investigated groups of patients. The possible mechanisms, by which Lf increases PN and PI of neutrophils, is discussed: 1) they may concern the antioxidative properties of Lf to block the iron ions in their catalytic inactive form or to take part as ferric-Lf in an oxidative-reduction processes on the plasma membrane and controlling transmembrane transport systems, 2) Lf decreases the negative surface charge and thus enhances the adherent ability of the PMN. Probably to this stimulated adherent ability dues the increased ingestion of bacteria in the presence of Lf, and 3) The "changed" membrane of PMN may have higher number receptors for Lf to bind more molecules of exogenous Lf. The increase of Lf binding which enhances the adherence and aggregation of neutrophils, facilitates the phagocytosis.

Topics: Adolescent; Adult; Antigens, Bacterial; Autoimmune Diseases; Humans; Hypersensitivity; Lactoferrin; Lymphocyte Activation; Male; Neutrophils; Phagocytosis; Staphylococcus aureus

The aim of this study was to determine whether allergic inflammation induces nasal hyperreactivity to bradykinin by enhancing neuronal responsiveness.. We compared the response to localized, unilateral nasal challenge with bradykinin in patients with perennial allergic rhinitis and nonallergic subjects, and in patients with seasonal allergic rhinitis challenged in and out of season. Weights of secretions from each nostril were recorded, and levels of albumin and lactoferrin in secretions recovered from each nostril were assayed. Contralateral administration of atropine (0.32 mg) was used to evaluate the role of cholinergic reflexes in nasal hyperresponsiveness to bradykinin.. In patients with symptomatic allergy, bradykinin induced greater symptom scores than in asymptomatic atopic or nonallergic control subjects. Moreover, bradykinin caused sneezing in a majority of patients with symptomatic allergy but in none of the asymptomatic atopic or nonallergic control subjects. Only patients with symptomatic allergy showed dose-dependent bilateral increases in secretion weights and levels of the serous glandular marker, lactoferrin. In contrast, bradykinin induced similar increases in ipsilateral, but not contralateral, levels of albumin in all patient populations. Atropine inhibited contralateral secretion and lactoferrin production (p < 0.05) in patients with symptomatic allergy.. The induction of sneezing and of atropine-inhibitable contralateral glandular secretion demonstrates that allergic inflammation causes nasal hyperreactivity to bradykinin, at least in part, by enhancing neuronal responsiveness.

Topics: Adult; Albumins; Allergens; Atropine; Bradykinin; Cross-Over Studies; Double-Blind Method; Female; Humans; Hypersensitivity; Immunoglobulin E; Inflammation; Lactoferrin; Male; Muscarinic Antagonists; Nasal Mucosa; Nasal Provocation Tests; Reflex; Rhinitis, Allergic, Perennial; Rhinitis, Allergic, Seasonal

Topics: Humans; Hypersensitivity; Lactoferrin; Lymphocyte Activation; Molecular Weight; Neutrophils; Skin; Skin Window Technique; Superoxides

Our previous skin chamber studies have shown prominent accumulation of viable neutrophils in human allergic skin reaction sites. To determine whether such neutrophils release components that may be pathogenic in allergic reactions, we have compared the patterns of release of five components: 1) lactoferrin, present in specific granules; 2) and 3) elastase and myeloperoxidase, present mainly in azurophilic granules; 4) lactic dehydrogenase, a cytosolic component generally released during cell damage; 5) histamine, present in mast cells and basophils but not in neutrophils. In 13 pollen-sensitive subjects we found that continuous antigen challenge for 5 h lead to a peak of histamine release into overlying skin chambers during the 1st h, followed by a plateau of low level histamine release over the succeeding 4 h. In contrast, there was no significantly increased released of lactoferrin or elastase during the first h, but significantly increased accumulation of these components at Ag challenge sites over the next 4 h. There was no significant difference at Ag vs buffer control sites in the levels of either myeloperoxidase or lactic dehydrogenase. The increased levels of lactoferrin and elastase at antigen challenge sites in the 2nd to 5th h were not simply a reflection of the greater numbers of neutrophils present in such sites because the levels of these components did not correlate significantly with the number of neutrophils in chamber fluids obtained from individual sites. However, such lactoferrin levels did correlate significantly with the amount of histamine released earlier during the 1st h of Ag challenge at individual sites. These findings suggest a selective in vivo release of neutrophil components in IgE-mediated human allergic skin reactions, possibly related in degree to earlier mast cell activation. Inasmuch as lactoferrin likely plays a role in reactive oxidants effects and elastase is a potent nonspecific protease, release of these agents could play a pathogenic role in late phase allergic reactions.

Topics: Adult; Histamine; Histamine Release; Humans; Hypersensitivity; L-Lactate Dehydrogenase; Lactoferrin; Lactoglobulins; Neutrophils; Pancreatic Elastase; Peroxidase; Skin

Topics: Arthritis, Rheumatoid; Collagen Diseases; Humans; Hypersensitivity; Lactoferrin; Lactoglobulins; Neutrophils; Phagocytosis; Staphylococcus aureus; Stimulation, Chemical