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Studies of the properties of immune complexes (IC) in the circulation, urine, and mesangium of IgA nephropathy (IgAN) patients have provided data relevant to the pathogenesis of this disease. IC contain predominantly polymeric IgA1 molecules which are deficient in galactose (Gal) residues on O-linked glycan chains in the hinge region (HR) of their heavy (H) chains. As a result of this aberrancy, a novel antigenic determinant(s) involving N-acetylgalactosamine (GalNAc) and perhaps sialic acid (SA) of O-linked glycans is generated and recognized by naturally occurring GalNAc-specific antibodies. Thus, IC in IgAN consist of Gal-deficient IgA1 molecules as an antigen, and GalNAc-specific IgG and/or IgA1 as an antibody. IgG antibodies to Gal-deficient IgA1 are probably induced by cross-reactive microbial antigens; they are present at variable levels not only in humans with or without IgAN but also in many phylogenetically diverse vertebrate species. Incubation of human mesangial cells with IC from sera of IgAN patients indicated that stimulation of cellular proliferative activity was restricted to the large (>800 kDa) complexes. These findings suggest that experimental approaches that prevent the formation of large Gal-deficient IgA1-IgG IC may be applied ultimately in an immunologically mediated therapy. Copyright © 2008 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Vitamin A supplementation during acute pneumonia has not improved recovery in most human clinical trials. We hypothesize that high vitamin A intake may decrease the production of T-helper type-1 (Th1) cytokines and thereby inhibit antiviral responses. Such decreases might impair recovery from viral respiratory infections. We thus examined the effect of three interventions on viral pneumonia: 1) a high level vitamin A [250,000 IU/kg diet or 75,000 retinol equivalents (RE)/kg], or 2) control diet (4000 IU/kg diet or 1200 RE/kg) given before and during infection, and 3) initiating the high level diet upon infection to simulate the adjuvant therapy used in clinical trials. No difference was seen among the interventions in severity of disease (weight loss, lung virus titers and survival). However, both the high level diet group and the group in which vitamin A was increased at the time of infection had greater salivary immunoglobulin (Ig)A responses (geometric means, 166 and 105 microg/L, respectively) than did the control group (59 microg/L) (P = 0.0019). In contrast, the serum IgG response was higher in the control group (324+/-158 mg/L) than in the high level group (225+/-95 mg/L) (P = 0.028), although it did not differ from the group in which the diet was changed upon infection (230+/-163 mg/L) (P = 0.084). The production of interferon-gamma (IFN-gamma), a Th1 cytokine, was lower in the high level diet group (median, 0.153 microg/L) compared with the control group (median, 0.839 microg/L) (P = 0.014), whereas the production of interleukin-10 (IL-10), a Th2 cytokine, was higher with the high level diet (median, 0.304 microg/L) than with the control (median, 0.126 microg/L) (P = 0.022). This change in the Th1/Th2 pattern was not sufficient to affect recovery from viral pneumonia but may account for the increased IgA and decreased IgG responses seen with high level dietary vitamin A in this study. These data reinforce the lack of utility of vitamin A in treating acute pneumonia in children and suggest that high dose vitamin A supplements may enhance Th2-mediated immune responses, which are particularly beneficial in the case of extracellular bacterial and parasitic infections and IgA-mediated responses to mucosal infections. [ABSTRACT FROM AUTHOR]

We analyzed the mechanism of human polymeric immunoglobulin receptor (pIgR) gene upregulation by tumor necrosis factor (TNF)-α. Northern blot analysis showed that the expression of pIgR mRNA was enhanced by TNF-α stimulation. This activation was completely inhibited by RNA polymerase or protein synthesis inhibitors, suggesting that the regulation of pIgR gene expression depends on de novo RNA and protein synthesis. Furthermore, the stimulation of pIgR mRNA by TNF-α was decreased by pyrrolidinedithiocarbamate and l-1-4′-tosylamino-phenylethyl-chloromethyl ketone, which are known nuclear factor (NF)-κB inhibitors. For further analysis of gene regulation, we cloned and sequenced the 1.5-kb 5′-flanking region of the pIgR gene. In the upstream region, we found two NF-κB-binding motifs (named κB1 and κB2 from the 5′ region). An electrophoretic mobility shift assay indicated that two components of the NF-κB/Rel family, p50 and p65, bound with higher affinity to the κB2 element than to the κB1 element. We also analyzed pIgR gene expression using reporter plasmids expressing the firefly luciferase gene. Stimulation by TNF-α significantly activated the pIgR gene promoter, as a 775-bp upstream region of the pIgR gene increased luciferase gene expression in cells treated with TNF-α. The activation of promoter activity by TNF-α was abolished when a mutation was inserted into κB1 or κB2. These data indicated that pIgR gene expression induced by TNF-α is transcriptionally regulated via activation of NF-κB. In addition, there is a possibility that another factor may act in concert with NF-κB. [ABSTRACT FROM AUTHOR]

The possibility that patients with IgA nephropathy (IgAN) might have abnormal IgA immune responses to immunogens commonly encountered at mucosal surfaces, resulting in the formation of circulating immune complexes (CIC), was examined. Since it is generally held that such increased IgA responses are characterized by detectable aberrancies in handling of IgA-containing CIC, IgAN patients and controls were given a large volume of bovine milk (after dietary deprivation of bovine antigens) and immune complex levels were measured over a period of 12 h. An assay based on binding of CIC containing C3 to solid-phase anti-C3 and subsequent development with isotype-specific antibody revealed no differences in responses of patients and controls with respect to IgG- and IgM-containing CIC. Although IgAN patients tended to have higher levels of IgA-containing CIC, there were no differences in response patterns when IgA CIC levels after ingestion of the milk stimulus were related to baseline levels. Polymorphonuclear leucocytes (PMNC), which bear surface receptors for IgA, were isolated from some subjects at the same times as the samples for CIC levels and examined by two-colour immunfluorescence for the coincident presence of IgA and milk antigens. In contrast to the data obtained in the CIC assays, these experiments revealed the simultaneous presence of IgA and two of three milk proteins in PMNC of IgAN patients but not controls. Follow-up experiments designed to assess more quantitatively the coincidental presence of IgA and milk antigens indicated no significant differences between patients and controls. However, milk proteins seemed to be more commonly associated with IgA in PMNC of IgAN patients, suggesting the presence of noncomplement-fixing IgA/antigen CIC after mucosal challenge of some IgAN patients. [ABSTRACT FROM AUTHOR]

The humoral and genital secretory immune response to chronic SIV infection was compared between female Rhesus macaques inoculated by i.v, or intravaginal routes. Total IgG levels in serum were 10- fold higher in SIV-infected animals when compared with uninfected controls. Vaginal washes from normal macaques contained predominantly IgA and lgG. while those from SI V-infected animals contained high levels of IgG. The SIV-infected animals had high Litres of SIV-specific IgG in serum. with lower hut detectable IgA and IgM responses. The genital secretory immune response to SIV was similar in intravenously and intravaginally inoculated animals, The anti-SIV response in the vaginal washes consisted mainly of lgG. Within the lamina propria of the reproductive tract of animals chronically infected with SIV there were essentially no IgA or lgG plasma cells and only a small number of IgM plasma cells, while two normal animals had large number animales of IgA plasma cells. These results suggest that. the mucosal immune system of the female reproductive tract is impaired in chronic SIV infection. [ABSTRACT FROM AUTHOR]

Human IgA occurs in multiple molecular forms (polymeric and monomeric) and two subclasses which show differential distribution between the mucosal and circulatory compartments of the immune system. However, the molecular form and subclass of specific IgA antibodies are influenced, especially during an immune response, by the type of antigen and duration of the response as well as by the route of exposure. These considerations question previously held notions that polymeric IgA and an increased representation of the IgA2 subclass among circulating antibodies or antibody-secreting cells signify their mucosal origin. Although the functional properties of different molecular forms and subclasses of IgA antibodies are incompletely understood, it appears that there is physiological benefit in the diversity of the IgA immune system. [ABSTRACT FROM AUTHOR]

Using modified ELISA and spot-ELISA. which permit the parallel determination of heavy chain subclass and the presence of covalently linked J chain, we analysed IgA found in cell culture supernatants or secreted by individual cells from peripheral blood, spleen, bone marrow, gingiva and synovial tissue, with respect to its polymeric or monomeric IgA form (pIgA, mIgA1 and IgA1 or lgA2 subclass. The ELISA for determination of J chain in tissue culture supernatants was specific and highly sensitive (detection limit in pg). The results demonstrated that IgA1-producing cells predominated in the tissues examined, and that J chain could he detected in association with the majority of IgA1 and lgA2 secreted by individual cells. With respect to the frequency of cells secreting polymeric, J chain-containing IgA, only 20-30% of cells from the bone marrow were engaged in the synthesis of pIgA. In other tissues the frequency of cells secreting pIgA1 and pIgA2 was considerably higher. Peripheral blood mononuclear cells secreting pIgA2 were easily inducible during stimulation with T cell-dependent pokeweed mitogen, whereas Epstein Barr virus-transformed cells secreted preferentially mIgA1. When the frequencies of pIgA-, pIgA1 - or pIgA2-secreting cells (determined by spot-ELISA technique) from different tissues were correlated with the proportion at pIgA to mIgA (and IgA subclasses) secreted in tissue culture supernatants, data obtained suggest that many individual IgA-producing cells could be engaged in simultaneous secretion of mIgA and pIgA. [ABSTRACT FROM AUTHOR]

The emergence of cells that produce IgG and IgA subclass antibodies to Bacteroides gingivalis (Porphyromonas gingivaiis) fimbriae and lipopolysaccharide (LPS) antigens was examined in mononuclear cells isolated from inflamed gingiva of different stages (slight, moderate or advanced) of adult periodontitis (AP). Antigen-specific IgM, IgG (including IgG1, IgG2, IgG3 and IgG4) and IgA (including IgAl and IgA2) producing cells were enumerated by the ELISPOT assay and were compared with total Ig-producing cells of each isotype or subclass. In advanced AP. the B. gingitalis fimbriae-specific IgG- and IgA-secreting cells represented 5% of total IgG- or IgA-secreting cells. while those from the moderate stage comprised approximately 1% of these two isotypes. Cells producing antibody specific for B. gingivalis LPS were observed at frequencies of 0.1% and 0.4% for IgG and IgA cells, respectively in the advanced stage. When IgG subclasses were analysed in moderate AP. the anti-fimbriae subclass responses were largely IgG1 (60%), followed by IgG2 (20%). lgG3(10%)and IgG4(10%), Fimbriae-specific IgG subclass responses were elevated in the advanced stage of AP, and IgG4 (40%) and IgG1 (30%) were dominant, followed by IgG3 (20%) and IgG2 (10%). IgA1 cells predominated in both the moderate and advanced stages, however a relative increase in IgA2 cells occurred in advanced AP, Mononuclear cells isolated from gingiva of AP patients did not contain cells producing antibody to antigens such as Fscherichia coli K235 LPS, cholera toxin or the hapten dinitrophenyl coupled lo bovine serum albumin. These results show that local IgG and IgA subclass responses occur to a protein antigen of a major periodontal disease (PD)- associated pathogen, B. gingivalis. and the increase in IgG4 and IgA2 responses may be associated with host protection. [ABSTRACT FROM AUTHOR]

The secretory immunoglobulin A (IgA) antibody response to infections of mucosal surfaces requires transport of IgA from the basal to apical surface of mucosal epithelial cells by a specific transport protein, the polymeric immunoglobulin receptor (pIgR). We have tested the hypothesis that the vitamin A metabolite all-trans retinoic acid (RA) is required for the regulation of pIgR expression by the cytokines interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) in HT-29 cells, a well-differentiated human epithelial cell line derived from a colonic carcinoma. pIgR expression is upregulated by IFN-gamma and IL-4 when HT-29 cells are grown in normal media, but this upregulation was significantly lower when cells were grown in vitamin A-depleted media. Treatment with RA at concentrations from 10(-9) to 10(-5) mol/L restored normal levels of pIgR expression. The percentages of cells expressing cell-surface pIgR after 24, 48 and 72 h of treatment with RA, IL-4 and IFN-gamma were 66 +/- 10, 90 +/- 5 and 92 +/- 1, respectively, significantly higher than the percentages seen without RA treatment, which were 32 +/- 2.3, 72 +/- 1.2 and 30 +/- 7, respectively. In addition, the intensity of fluorescence of pIgR-positive cells was significantly higher in the RA-treated cultures than in the cultures without RA treatment. Similarly, pIgR mRNA levels (adjusted for beta-actin mRNA levels) in RA-supplemented cultures were 404, 105 and 949% higher at 24, 48 and 72 h, respectively, than were pIgR mRNA levels in identical cultures grown in the absence of RA. These data indicate that RA strongly interacts with IL-4 and IFN-gamma to regulate pIgR expression in HT-29 cells, suggesting that vitamin A may be required for proper in vivo regulation of IgA transport in response to mucosal infections. [ABSTRACT FROM AUTHOR]

Murine T cells and T-cell lines express receptors for the Fc of IgA (Fc α R); however, their molecular properties remain to be elucidated. In the present study, we examined three candidate molecules for IgA-binding receptors including Fc α R, β-galactosyltransferase (β-GT) and anti-secretory component (SC) reactive proteins (ASCP) expressed on T cells which might participate in the binding of different molecular forms of IgA. T-cell lines derived from CD4+ T cells of mouse Peyer's patches (PP) (designated PPT 4-6 and PPT 4-16) and from cloned PP T helper (Th) cell lines (ThHA1 #9 and #10) bound both monomeric and dimeric IgA (mIgA and dIgA), while the fusion partners (BW 5147 and R1.1) did not. In contrast, both Fc α R+ and Fc α R- cell lines bound to high molecular weight polymeric or aggregated IgA (pIgA). All cell lines reacted with a monoclonal anti-β-GT (MoAb) and β-GT enzyme activity was associated with the cell lysates and membrane fractions of all cells tested. The anti-β-GT MoAb stained a 47-kDa band on immunoblots which was identical to that seen with native enzyme. mRNA analysis with β-GT cDNA showed that all cell lines constitutively produced enzyme-specific mRNA. Both Fc α R+ T cells and Fc α R- control cell lines showed cell surface specific β-GT activity. This is the first study which shows that mouse T cells produce β-GT. However, Fc α R and β-GT appear to be separate receptors, because Fc α R+ T cells bound mIgA and dIgA, and this treatment did not affect staining with biotinylated anti-β-GT MoAb. Further, preincubation of the Fc α R+ cells with anti-β-GT MoAb did not block mIgA binding. However, the anti-β-GT MoAb partially blocked binding of pIgA to both Fc α R+ and Fc α R- T cells, suggesting that β-GT may be a receptor for pIgA. Others have shown that T cells may bind IgA through a receptor serologically related to SC. We found that antibodies both to human SC and to rat SC specifically bound to both Fc α R+ and Fc α R- T cells. Further, a 72-kDa band was detected when cell membrane fractions were analysed with these antisera (ASCP) by solid phase immunoisolation technique and immunoblot analysis. The ASCP is not an IgA-binding receptor, since anti-SC did not block either mIgA or pIgA binding. Further, the effects of proteolytic enzymes were different on these three IGA-binding molecule candidates. Fc%alpha;R and ASCP were shown to be sensitive to pronase proteolytic degradation, but were resistant to trypsin and trypsin/EDTA treatements. In contrast, β-GT was sensitive to both pronase and trypsin treatements. We conclude that multiple IGA receptors are present on mouse T cells, and include those which bind to mIGA or dIgA (FcαR) as well as those wihich bind to pIgA (β-GT). Further, ASCP is also present on mouse T-cells lines, but its role in IgA binding to T cells remains to be further determined. [ABSTRACT FROM AUTHOR]

Because of similarities between the human and monkey immune systems, we considered the monkey a suitable model for studies on the catabolism of various molecular forms of IgA, for which little information is available. The residualizing label dilactitol-[125I] tyramine was coupled to monkey (Macaca fuscata) IgA and IgG, as well as to human monomeric and polymeric myeloma IgA1 and IgA2 proteins. When labelled proteins were injected intravenously into monkeys, the non-metabolizable radioiodinated tracer accumulated at the cellular site of protein degradation, allowing identification of the catabolic sites. To determine the uptake of injected proteins by various tissues, monkeys were sacrificed 6-7 days after injection of labelled proteins, when blood-associated radioactivity was ⩽ 10% of the injected dose, as measured by plasma clearance. When monkey or human monomeric IgA, as well as human polymeric IgA, irrespective of subclass, was administered to monkeys, the liver showed the greatest tissue uptake relative to total dose injected and to organ weight, and the highest acid soluble radioactivity (degraded protein). Although both hepatocytes and non-parenchyma liver cells were involved in IgA uptake, the hepatocytes were more active. Therefore, it appears that the liver is the major site of uptake and catabolism of IgA in monkeys and possibly in humans. [ABSTRACT FROM AUTHOR]

Polymeric microspheres were evaluated as an oral antigen delivery system for immunization with influenza virus. The immune responses obtained were compared after either oral or systemic immunization of BALB/c mice using purified, formalin-inactivated influenza virus type A/H3N2, either encapsulated in biodegradable and biocompatible microspheres or free in solution. The immunogenicity of formalin-treated influenza vaccine was preserved during the microencapsulation process, and the microencapsulated antigen induced protective immune responses after systemic immunization that were equal to or higher than those induced by conventional vaccine. When administered orally to primed animals, microencapsulated antigen induced levels of anti-influenza antibodies in saliva that were higher than and in serum that were comparable to those obtained by systemic immunization. Furthermore, oral booster immunization provided virtually complete protection of animals challenged with live virus. [ABSTRACT FROM AUTHOR]

An abstract of the research paper "Improved systemic and mucosal antibody responses with a CCR10 ligand adjuvant," by N. Hutnick and colleagues is presented.