Gerco den Hartog

Successive immunoglobulin and cytokine expression in the small intestine of juvenile chicken

The intestinal mucosa is of major importance for immune development. To further study the ontogeny of avian mucosal immunity, mRNA levels of IgM, IgY and IgA, the polymeric immunoglobulin receptor (pIgR) and a number of cytokines were determined at different ages in jejunum and ileum of non-immunized healthy juvenile layer chickens. Immunoglobulin genes were successively expressed in jejunum and ileum. IgM expression was maximal in week 1, IgY expression peaked in week 5, and IgA expression was most dominant after week 7 post hatch. PIgR gene expression was relatively low in the first 2 weeks post hatch, but increased thereafter. Generally, increased expression levels of IL-1, IL-10, IL-12p40, iNOS and interferon-γ mRNA levels were found between days 14-42 as compared to days 3 and 49-70 post hatch (p<0.05). Correlation was found between IgA and IL-10, TGF-β and IFN-γ expression levels on days 21, 28 and 35. Cytokine mRNA expression levels decreased to basal levels between 49 and 70 days post hatch, whereas IgA reached its maximum levels in this period. Based on the current results, we hypothesize that chicken sIgA, as mammalian sIgA, may contribute to the maintenance of intestinal homeostasis.

Modulation of human immune responses by bovine interleukin-10.

Cytokines can be functionally active across species barriers. Bovine IL-10 has an amino acid sequence identity with human IL-10 of 76.8%. Therefore, the aim of this study was to evaluate whether bovine IL-10 has immunomodulatory activities on human monocytes and dendritic cells. Peripheral blood monocytes were isolated from healthy donors, and used directly or allowed to differentiate to dendritic cells under the influence of IL-4 and GM-CSF. Recombinant bovine IL-10 inhibited TLR induced activation of monocytes, and dose-dependently inhibited LPS-induced activation of monocyte-derived DCs comparable to human IL-10. By using blocking antibodies to either bovine IL-10 or the human IL-10 receptor it was demonstrated that inhibition of monocyte activation by bovine IL-10 was dependent on binding of bovine IL-10 to the human IL-10R. These data demonstrate that bovine IL-10 potently inhibits the activation of human myeloid cells in response to TLR activation. Bovine IL-10 present in dairy products may thus potentially contribute to the prevention of necrotizing enterocolitis and allergy, enhance mucosal tolerance induction and decrease intestinal inflammation and may therefore be applicable in infant foods and in immunomodulatory diets.

Specificity and Effector Functions of Human RSV-Specific IgG from Bovine Milk.

Background Respiratory syncytial virus (RSV) infection is the second most important cause of death in the first year of life, and early RSV infections are associated with the development of asthma. Breastfeeding and serum IgG have been shown to protect against RSV infection. Yet, many infants depend on bovine milk-based nutrition, which at present lacks intact immunoglobulins. Objective To investigate whether IgG purified from bovine milk (bIgG) can modulate immune responses against human RSV. Methods ELISAs were performed to analyse binding of bIgG to human respiratory pathogens. bIgG or hRSV was coated to plates to assess dose-dependent binding of bIgG to human Fcγ receptors (FcγR) or bIgG-mediated binding of myeloid cells to hRSV respectively. S. Epidermidis and RSV were used to test bIgG-mediated binding and internalisation of pathogens by myeloid cells. Finally, the ability of bIgG to neutralise infection of HEp2 cells by hRSV was evaluated. Results bIgG recognised human RSV, influenza haemagglutinin and Haemophilus influenza. bIgG bound to FcγRII on neutrophils, monocytes and macrophages, but not to FcγRI and FcγRIII, and could bind simultaneously to hRSV and human FcγRII on neutrophils. In addition, human neutrophils and dendritic cells internalised pathogens that were opsonised with bIgG. Finally, bIgG could prevent infection of HEp2 cells by hRSV. Conclusions The data presented here show that bIgG binds to hRSV and other human respiratory pathogens and induces effector functions through binding to human FcγRII on phagocytes. Thus bovine IgG may contribute to immune protection against RSV.

Ontogeny of the avian intestinal immunoglobulin repertoire: Modification in CDR3 length and conserved VH-pseudogene usage

Immunoglobulins play an important role in maintenance of mucosal homeostasis in the gut. The antigen binding specificity of these immunoglobulins depends for a large part on the hypervariable CDR3 region. To gain knowledge about isotype-specific development of the CDR3 repertoire we examined CDR3 spectratypes at multiple time points between 4 and 70 days post hatch. In order to identify clonal expansions deviation from the normal distribution (SS) and the average CDR3 length was calculated. IgA-CDR3 regions were studied in more detail by DNA sequence analysis at day 7 and 70 and preferential VH pseudogene usage was estimated. The SS of CDR3 repertoires of the IgM, IgG and IgA isotypes successively increased, but for each isotype this increase was transiently. The length of the CDR3 regions decreased with age for IgM becoming similar to the CDR3 length of IgA at day 70. The IgA- and IgG-CDR3 lengths did not change with age. On average, the CDR3 length of IgA was the shortest. IgA CDR3 sequences were similar between animals aged 7 and 70 days. A limited number of pseudogenes was used, and no differences in pseudogene usage were observed between animals aged 7 and 70 days. Of the identified VH pseudogenes, half of the sequences used VH15, whilst a number of the pseudogenes were not used at all. We conclude that CDR3 spectratype profiles change during aging, whilst at the CDR3-sequence level, variation in VH pseudogene usage for ileal IgA is limited suggesting conservation during ontogeny.

Induction of salivary antibody levels in Dutch adolescents after immunization with monovalent meningococcal serogroup C or quadrivalent meningococcal serogroup A, C, W and Y conjugate vaccine.

Background Meningococcal infection starts with colonisation of the upper respiratory tract. Mucosal immunity is important for protection against acquisition and subsequent meningococcal carriage. In this study, we assessed salivary antibody levels against meningococcal serogroup A (MenA), W (MenW) and Y (MenY) after vaccination with a quadrivalent MenACWY conjugated vaccine. We also compared salivary meningococcal serogroup C (MenC) antibody levels after monovalent MenC and quadrivalent MenACWY conjugated vaccination. Methods Healthy participants, who had received MenC conjugate vaccine between 14 months and 3 years of age, received a (booster) MenC or MenACWY vaccination at age 10–15 years. MenA-, MenC-, MenW- and MenY-polysaccharide (PS) specific IgG and IgA levels in saliva and serum and PS specific secretory component levels in saliva were measured using the fluorescent-bead-based multiplex immunoassay. Results MenACYW vaccination increased salivary PS-specific IgA (2-fold) and IgG levels(>10-fold) for MenA, MenY, and MenW. After one year, salivary IgA levels had returned to baseline levels. Both vaccines induced an increase in salivary MenC-PS specific IgA (>3-fold) and IgG (>100-fold), with higher levels after MenC as compared to MenACWY vaccination. The antibody decay rate of MenC in saliva between one month and one year was similar for both vaccines. The overall correlation between serum and saliva IgA levels was low (R = 0.39, R = 0.58, R = 0.31, and R = 0.36 for MenA, MenC, MenW and MenY, respectively). Serogroup-PS specific IgG levels between serum and saliva correlated better (R ranged from 0.51 to 0.88). Conclusions Both primary (MenA, MenY, and MenW) and booster (MenC) parenteral meningococcal conjugate vaccination induced high salivary antibody levels. The strong correlation for MenC, MenW and MenY between saliva and serum IgG levels indicates that saliva might be used as a reliable tool to measure vaccine responses after both primary and booster meningococcal vaccination.

BAFF augments IgA2 and IL‐10 production by TLR7/8 stimulated total peripheral blood B cells

Class‐switching of B cells to IgA can be induced via both T‐cell‐dependent and T‐cell‐independent mechanisms. IgA is most predominantly produced mucosally and is important for combating infections and allergies. In contrast to mice, humans have two forms of IgA; IgA1 and IgA2 with diverse tissue distribution. In early life, IgA levels might be sub‐optimal especially during the fall season when bacterial and viral infections are more common. Therefore, we investigated using human B cells whether T‐cell‐independent factors ‐promoting cell survival, class switching and immunoglobulin secretion‐ BAFF, APRIL, IL‐10 and retinoic acid can boost IgA production in the context of viral or bacterial infection. To this end total and naive peripheral blood B cells were stimulated with these factors for 6 days in the presence or absence of TLR7/8 agonist R848 (mimicking viral infection) or TLR9 agonist CpG‐ODN (mimicking bacterial infection). We show that BAFF significantly augments IgA2 production in TLR7/8 stimulated mature, but not naïve B cells. In addition, BAFF augments IL‐10 production and viability in TLR7/8 and TLR9 stimulated mature B cells. These data warrant further investigation of its role in immune regulation both in the periphery and mucosal tissues in early life or during disease.

An alternative inhibition method for determining cross-reactive allergens

Abstract Background: Inhibition assays are an useful tool to identify the allergen of primary sensitization of cross-reactive allergens. Classical ELISA-based inhibition assays are limited by both the availability of commercial standardized allergen extracts and the experience and knowledge needed for making home-made extracts. Moreover the direct comparison of the inhibition ELISAs outcomes between different laboratories is difficult because of different sources of used allergen extracts and a number of methodological variations. Therefore, we propose a novel ImmunoCap (Phadia, Thermofisher Scientific) based immunoinhibition method with the use of commercially available Caps as the allergen source. Methods: The novel ImmunoCap based immunoinhibition method was developed and tested with sera from patients with a well-known cross-reactive sensitization for fig (Ficus carica) and ficus (Ficus benjamina). Results were compared with a classically applied inhibition method, i.e. addition of homemade allergen extract to patient serum. Results: The amount of allergens (fig and ficus extracts) needed to reach a similar degree of inhibition was comparable for both inhibition methods. Conclusions: The ImmunoCap based inhibition assay, in addition to classical inhibition methods, is a valuable tool as the ImmunoCap analyzer and commercial allergens (Caps) are more widely available which makes the outcomes of inhibition tests comparable between different laboratories. Furthermore, in the ImmunoCap inhibition method the same protein source is used for both the inhibition of sIgE and sIgE measurement, which might be even more relevant when multiple cross-reactive allergens are tested.

Associate Editor Hirohisa Saito

Prof. Hirohisa Saito, Deputy Director of the National Research Institute for Child Health and Development, serves as a board member of the Japanese Society of Allergology (JSA), Editor in Chief of Allergology International (official journal of the JSA) and Associate Editor of the Journal of Allergy and Clinical Immunology . He graduated from Jikei University School of Medicine in 1977 and started his career as a Pediatrician. After receiving his PhD, he taught immunology, especially mast cell biology, at the Johns Hopkins University, under the supervision of Prof. Teruko Ishizaka, from 1986 until 1988. In 1996, after serving as a clinical allergy specialist, he was appointed as Director of the Department of Allergy and Immunology, National Children’s Medical Research Center. In 2002, his Institute was unified and renamed as National Research Institute for Child Health and Development. Since 2003, he has been Professor of Pediatrics at Jikei University, Toho University and Juntendo University. He was concurrently serving as Leader of the Allergy Transcriptome Unit at the Research Center for Allergy and Immunology, RIKEN, from 2002 until 2006. In 2010, he was promoted to Deputy Director of the Research Institute for Child Health and Development. In 2013, he was elected President of the Japanese Society of Allergology. Published online: September 5, 2013

Associate Editor Andreas Radbruch

A biologist by education, Andreas Radbruch did his PhD at the Genetics Institute of the Cologne University with Klaus Rajewsky. He later became Associate Professor there and was a visiting scientist with Max Cooper and John Kearney at the University of Alabama, Birmingham. In 1996, Andreas Radbruch became Director of the German Rheumatism Research Center in Berlin, a Leibniz Institute, and in 1998, Professor of Rheumatology at the Charité Medical Center and Humboldt University of Berlin. Andreas Radbruch has been President of the German Society for Rheumatology, the German Society for Immunology and is incoming President of the International Society for Advancement of Cytometry (ISAC). He serves on a number of advisory and editorial boards and is a fellow of many academic organizations. He is Editorial Chair of the European Journal of Immunology . Most recently, he was awarded the Carol Nachman Prize and an advanced research grant of the European Research Council. Andreas Radbruch has authored more than 250 original publications on immunological memory, antibody class switching, T and B lymphocyte differentiation, cytometry and cell sorting. His research group described the organization of memory plasma cells and memory T helper (Th) lymphocytes in bone marrow and identified memory plasma cells secreting pathogenic antibodies as novel target in chronic immune-mediated diseases. Andreas Radbruch demonstrated, by targeted mutagenesis, that antibody class switch recombination in activated B lymphocytes is targeted to distinct switch regions by transcription. His group contributed significantly to our current understanding of Th1 and Th2 cytokine memory, its imprinting and plasticity and, more recently, has identified critical molecular adaptations of Th effector memory cells to chronic inflammation. Radbruchs group developed the MACS technology and the cytometric secretion assay. Published online: September 5, 2013