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This study aimed to elucidate the role of polymeric immunoglobulin receptor (pIgR) autoantibodies in the pathogenic progression of biliary atresia (BA). The presence and levels of plasma pIgR autoantibodies, pIgR antigen expression, and B cell counts were assessed in liver tissues. Serum extracellular vesicles (EVs) were isolated, quantified, and characterized. The functional roles of EVs enriched with pIgR autoantibodies in biliary injury were investigated. Infants diagnosed with BA exhibited significantly elevated levels of plasma pIgR autoantibodies, which positively correlated with hepatic inflammation. The expression levels of pIgR autoantibodies demonstrated high accuracy in distinguishing BA from non-BA controls. Notably, the presence of pIgR antigens was specifically observed in cholangiocytes and was associated with an increased number of CD27+ memory B cells within the liver tissue. Furthermore, the concentration of pIgR autoantibodies was found to be higher in EVs derived from BA patients compared to those from control subjects. EVs enriched with pIgR autoantibodies induced biliary injury potentially through activation of the extracellular signal-regulated kinase (ERK) pathway. Our findings suggest that pIgR autoantibody may serve as a potential biomarker for differentiating infants with BA from those without it. Additionally, these results indicate that EVs enriched with pIgR autoantibody could play a significant role in the underlying pathogenesis of BA. • Blood polymeric immunoglobulin receptor (pIgR) autoantibody levels are significantly elevated in infants with biliary atresia (BA), correlating positively with liver injury severity. • The blood pIgR autoantibody accurately differentiates between biliary atresia (BA) and non-BA cases. • Cholangiocytes in BA patients predominantly express the pIgR antigen, accompanied by increased CD27+ memory B cells. • Elevated pIgR autoantibody levels are present in circulating extracellular vesicles (EVs) from BA patients. • PIgR autoantibody-enriched EVs induce biliary injury by suppressing the ERK pathway. [ABSTRACT FROM AUTHOR]

To identify and evaluate differentially expressed plasma proteins in biliary atresia (BA), we performed plasma proteome profiling using liquid chromatography with tandem mass spectrometry (LC-MS/MS) in 20 patients with BA and 10 control children. Serological assays validated the most significant and highly upregulated proteins in a cohort of 45 patients and 15 controls. Bioinformatics tools were used for functional classification and protein-protein interactions of differentially expressed proteins (DEPs). Of 405 proteins detected in patients and 360 in controls, 242 proteins, each with ≥2 unique peptides (total of 3230 peptides), were common in both groups. Compared to controls, 90 proteins in patients were differentially expressed and were dysregulated. Twenty-five were significantly upregulated with polymeric immunoglobulin receptor (PIgR), galectin-3-binding protein (Gal-3BP), complement C2, the most prominent, and 15 had low expression. The bioinformatic analysis revealed functional interaction between DEPs and their role in an inflammatory immune response. Enzyme immunoassay for PIgR and Gal-3BP in patients' plasma showed their levels raised significantly (p = 0.0021 and p = 0.0369, respectively). The PIgR and Gal-3BP are novel proteins upregulated in BA and may be tested further for their utility as potential circulating disease biomarker(s). The study shows that plasma PIgR and GAL-3BP levels are significantly raised in infants with BA within the first 3 months of life. If tested in a larger cohort, these proteins may be found to have their diagnostic potential and utility as disease biomarkers. The study also provides valuable information on the involvement of several DEPs in innate immune response, chronic inflammation, and fibrosis. This strengthens the hypothesis that the immune-mediated inflammatory processes are responsible for the progressive nature of BA. [Display omitted] • Biliary atresia (BA) is a lethal infancy disorder requiring multiple diagnostic modalities to reach a diagnosis; hence there is a continuous search for a reliable biomarker for BA. • Of 405 differentially expressed proteins identified, the up and downregulation of 25 and 15 proteins were statistically significant compared to controls. • The bioinformatic analysis revealed functional interaction between most of these proteins and their link with immune response and inflammation. • PIgR and Gal-3BP are novel proteins upregulated in BA and may be tested for their utility as potential circulating disease biomarker(s). [ABSTRACT FROM AUTHOR]

To observe the effect of Salmonella enteritidis (SE) -induced inflammation on pIgR expression in jejunum and ileum. Salmonella enteritidis was orally administered to 7-day old Hyline chicks, which were killed after 1d,3d,7d and 14d. The mRNA expression of TLR4,MyD88,TRAF6,NF-κB, and pIgR was detected by real-time RT-PCR, and pIgR protein was detected by Western blotting. The TLR4 signaling pathway was activated, the mRNA expression of the pIgR in jejunum and ileum was increased, and pIgR protein in jejunum and ileum was up-regulated by SE. In SE-treated chicks,the pIgR in jejunum and ileum was up-regulated on mRNA,and protein level,associated with activation of the TRL4-mediated MyD88/TRAF6/NF-κB signaling pathway, which identifies this as a novel pIgR-related pathway to TLR4 activation. [ABSTRACT FROM AUTHOR]

Background/objectives Polymeric immunoglobulin receptor (pIgR) traffics Immunoglobulins (IgA and IgM) through epithelial cells in normal mucosae but neither are expressed in the normal pancreas. Recent work from our laboratory suggested pIgR may be upregulated in pancreatic ductal adenocarcinoma (PDAC). Our aim was to assess the role of pIgR in human PDAC. Methods pIgR expression was manipulated (siRNA and shRNA) in cell lines to evaluate its subsequent effect on cell behaviour in 2D assays as well as 3D organotypics models. Tissue Microarrays of 88 patients with PDAC were analysed after pIgR, αSMA, E-Cadherin and Picrosirius Red staining to assess their role as a combined bio-marker panel. Results Cytokines such as interleukin 4 (IL4) and Tumour Necrosis Factor (TNFα) could not modulate pIgR expression in PDAC cell lines despite this effect being seen in other studies. Down-regulation in pIgR expression in Capan1 cancer cell line resulted in reduction of cellular proliferation, adhesion and migration in 2D assays. In 3D physiomimetic organotypic models, pIgR downregulation resulted in reduced cancer cell invasion, alteration of apico-basal polarity and diminished stromal activity. In human PDAC, decreased E-cadherin expression correlates with increased pIgR expression through pancreatic intra-epithelial neoplasia (PanIN) progression. In combination with enhanced stromal indices (α-smooth muscle action (SMA) and Picrosirius red), low pIgR scores had a trend towards better survival. Conclusion pIgR may be involved in PDAC progression and may be linked stromal activity. Further work on its precise role is mandated in in vivo models, to understand its influence on cancer progression. [ABSTRACT FROM AUTHOR]

The improvement of disease resistance is vital in aquaculture as diseases are one of the serious threats to successful aquaculture. Understanding of molecular mechanisms underlying disease resistance is important in improving resistance against diseases. The pIgR (polymeric immunoglobulin receptor) gene plays an important role in the mucosal immunity of fish. However, not much is known about its roles in inhibiting pathogens in aquaculture species. Here, the pIgR gene from Asian seabass (Lates calcarifer) was characterized, and its functions on the iridovirus infection were examined. LcapIgR consisted of a 2683 bp ORF, a 69 bp 5' UTR and a 1597 bp 3' UTR. qRT-PCR revealed that LcapIgR was expressed in all 11 organs examined, being the highest in the heart, followed by intestine, kidney, and eye. Upon a SGIV (Singapore grouper iridovirus) challenge of the fish, the expression of LcapIgR in kidney increased significantly. Overexpression of LcapIgR in an Asian seabass cell line reduced iridovirus entry into cells, and significantly lowered the transcription and replication of the gene encoding for the iridovirus major capsid protein. These results suggest that LcapIgR play an important role in repressing the pathogenic activity of iridovirus. In addition, two SNPs were identified in the 3' UTR of LcapIgR. Genotyping of both SNPs in survival and dead Asian seabass after challenging with SGIV revealed that both SNPs were possibly associated with resistance to SGIV. Therefore, both SNPs may play a useful role in the selection of fish that are resistant against iridovirus at the fingerling stage. • The LcapIgR gene was characterized in the Asian seabass. • LcapIgR was expressed in all tissues, being the highest in the heart, intestine, eye and kidney. • Challenging with iridovirus increased the expression of LcapigR in the kidney. • Overexpression of LcapIgR lowered the transcription and replication of iridovirus. • Two SNPs in the 3' UTR of LcapigR were associated with iridovirus resistance. [ABSTRACT FROM AUTHOR]

Polymeric immunoglobulin receptor (pIgR) plays an important role in mucosal immune systems. Secretory immunoglobulin A, composed of secretory component of pIgR and a dimeric form of immunoglobulin A, is secreted on mucosal surfaces and serves as a biological defense factor. pIgR gene expression is reportedly induced by activation of the transcription factor nuclear factor (NF)-κB. On the other hand, secretory leukocyte protease inhibitor (SLPI) is a glycoprotein that functions as a serine protease inhibitor. In alveolar epithelial cells, SLPI increases the level of IκBβ, which indicates that it is an inhibitor of NF-κB at the protein level. Taken together, SLPI may regulate pIgR expression; however, the specific mechanism by which this occurs is unclear. Therefore, the aim of this study was to elucidatethe influence of SLPI on pIgR expression.SLPI and pIgR localized in goblet cells and ciliated epithelial cells of the gastrointestinal tract, respectively. No cells were detected in which SLPI and pIgR were co-expressed. In addition, recombinant human SLPI stimulation of an epithelial cell line (HT-29) decreased the pIgR expression. The pIgR expression was also higher in SLPI-deficient Ca9-22 cells than in wild-type Ca9-22 cells. Furthermore, a luciferase assay using a NF-κB reporter plasmid and real-time RT-PCR analysis indicated that when SLPI was present, the transcriptional activity of NF-κB protein was suppressed, which was accompanied by anincrease in the protein, but not the mRNA,expression of IκBβ. These results demonstrate that SLPI down-regulates pIgR expression through the NF-κB signaling pathway by inhibiting degradation of IκBβ protein. [ABSTRACT FROM AUTHOR]

Wilson's disease is a rare inherited disorder of copper metabolism, manifesting hepatic, neurological and psychiatric symptoms. Early diagnosis is often unfeasible and a unique diagnostic test is currently inapplicable. We performed the qualitative/quantitative characterization of the salivary proteome/peptidome of 32 Wilson's disease patients by an integrated top-down/bottom-up approach. Patients exhibited significant higher levels of S100A9 and S100A8 proteoforms, and their oxidized forms with respect to controls. Oxidation occurred on methionine and tryptophan residues, and on the unique cysteine residue, in position 42 in S100A8, and 3 in S100A9, that generated glutathionylated, cysteinylated, sulfinic, sulfonic, and disulfide dimeric forms. Wilson's disease patient saliva showed high levels of two new fragments of the polymeric immunoglobulin receptor, and of α-defensins 2 and 4. Overall, the salivary proteome of Wilson's disease patients reflected oxidative stress and inflammatory conditions characteristic of the pathology, highlighting differences that could be useful clues of disease exacerbation. [ABSTRACT FROM AUTHOR]

Intermittent fasting prolongs the lifespan and unlike intense stress provides health benefits. Given the role of the immunoglobulin A (IgA) in the intestinal homeostasis, the aim of this study was to assess the impact of intermittent fasting plus intense stress on secretory IgA (SIgA) production and other mucosal parameters in the duodenum and ileum. Two groups of six mice, with intermittent fasting or fed ad libitum for 12 weeks, were submitted to a session of intense stress by a bout of forced swimming. Unstressed ad libitum fed or intermittently fasted groups were included as controls. After sacrifice, we evaluated intestinal SIgA and plasma adrenal hormones, lamina propria IgA+ plasma-cells, mRNA expression of polymeric immunoglobulin receptor, α- and J-chains in the liver and intestinal mucosa, as well as pro- (tumor necrosis factor-α, interleukin-6 and Interferon-γ) and anti- (interleukin-2, -4, -10 and transforming growth factor-β) inflammatory cytokines in mucosal samples. Under intense stress, intermittent fasting down- or up-modulated the levels of most parameters in the duodenum and ileum, respectively while up-regulated corticosterone levels without affecting epinephrine. Our data suggest intermittent fasting plus intense stress elicited neuroendocrine pathways that differentially controlled IgA and pIgR expression in duodenum and ileum. These findings provide experimental foundations for a presumable impact of intermittent fasting under intense stress on the intestinal homeostasis or inflammation by triggering or reducing the IgA production in ileum or duodenum respectively. [ABSTRACT FROM AUTHOR]

Stress stimuli affect the immune system of the mucosa, and in particular IgA secretion. It is well documented that intense psychological and physical stress can increase susceptibility to infection by diverse pathogens in the upper respiratory tract. Our workgroup reported that chronic stress caused by immobilization elicits a decrease in nasal IgA levels in mice. Here, we explore how acute stress (caused by 4 h of immobilization) affects IgA secretion in the nasal mucosa, and the possible role of the sympathetic nervous system in this effect. Nine-week-old male CD1 mice were divided into four groups: control, chemical sympathectomy (with 6-OHDA) and treatment with nadolol (5 mg/kg) or phentolamine (15 mg/kg). All these groups were subdivided into stressed and unstressed animals. The parameters evaluated included plasma corticosterone and epinephrine (only in control groups), SIgA levels (by ELISA) and SIgA expression (by Western Blot) in nasal fluid, percentage of IgA + plasma cells, and mRNA expression of heavy alpha chain, pIgR, TNFα and TGFβ in nasal mucosa. Acute stress reduced the percentage of IgA + cells while increasing the levels of IgA, the two hormones, and the mRNA expression of heavy alpha chain, pIgR, TNFα and TGFβ, which resulted in greater synthesis and transport of IgA. The treatments with 6-OHDA and α- and β-adrenergic receptor blockers suggest that sympathetic innervation by both types of adrenergic receptors is important for the control of SIgA secretion in nasal mucosa during acute stress. The increase in this parameter depended on the cytokines involved in IgA synthesis and transport. [ABSTRACT FROM AUTHOR]

As mucosal barriers in fish are the main sites where pathogens are encountered, mucosal immunity is crucial to avoid infection in the aquatic environment. In teleost fish, immunoglobulins are present in gut, gill and skin mucus, although not in the same amounts as in higher vertebrates. In mammals, the poly-Ig receptor (pIgR) is synthesized in epithelial cells and mediates the active transport of poly-immunoglobulins (pIgs) across the epithelium. During transport, a component of the pIgR, the secretory component (SC), is covalently bound to pIgs secreted into the mucus providing protection against proteases and avoiding degradation. The teleost pIgR gene does not show synteny to higher vertebrates, the overall structure of the protein is different (comprising two Ig domains) and its functional mechanisms remain unclear. The J-chain which is essential for pIgR-mediated transport of IgA and IgM in higher vertebrates is absent in teleost fish. The aim of the present study was to characterize the ballan wrasse (Labrus bergylta) pIgR and use it as a marker for further studies of mucosal immunity in this species. The pIgR gene was unambiguously identified. Unexpectedly, reverse transcription real time PCR (RT-qPCR) revealed highest abundance of pIgR mRNA in liver and significantly lower expression in mucosal organs such as foregut, hindgut, and skin. In situ hybridization showed pIgR-positive cells dispersed in the lamina propria while it was undetectable in epithelial cells of foregut and hindgut of ballan wrasse. A similar pattern was observed in Atlantic salmon. Liquid Chromatography-Mass Spectrometry (LC-MS/MS) analysis of IgM enriched mucus samples from gut, gill, skin, and bile gave relatively few matches to wrasse pIgR. Notably, the matching peptides were from the transmembrane (TM) and cytoplasmatic (Cy) region as well as the putative SC, indicating leakage from lysed cells rather than covalent bonds between IgM and SC. Altogether, the results indicate that pIgR has another (or at least an additional) function in wrasse. Another pIgR-like molecule (pIgRL) in ballan wrasse (comprising three Ig domains) was analyzed to see if this could be an alternative functional pIgR homolog. However, the presence of pIgRL mRNA in blood leukocytes and a relatively high expression in immune organs like spleen and head kidney pointed to a receptor function on a circulating leukocyte population. As significant amounts of IgM were found in bile of ballan wrasse further studies should consider the hepato-biliary route regarding IgM delivery to the gut lumen. • The ballan wrasse pIgR counterpart has been characterized. • The abundance of pIgR mRNA was highest in liver and low in mucosal organs such as gut and skin. • Gut pIgR mRNA was present in cells in the lamina propria but undetected in the gut epithelium. • In situ hybridization revealed similar results in ballan wrasse and Atlantic salmon. • A significant amount of IgM was present in bile of ballan wrasse. [ABSTRACT FROM AUTHOR]

Abstract: Synthetic peptides incorporating the complementarity determining region 2 (CDR2)-like loop of domain 1 of human polymeric immunoglobulin receptor (hpIgR-D1) were investigated for their interactions with human Immunoglobulin-M (hIgM) and other hIgs. pep14, a 14mer peptide, emerged as a unique biomimetic ligand with specificity to interact with hIgM and negligible affinity for hIgG, hIgE, hIgA1 and bovine serum albumin. Surface plasmon resonance-based binding studies between immobilized pep14 and fragments of hIgM molecule, namely Fc5μ and Fab, suggested that pep14 was binding to a motif in the constant domain 2 of hIgM. Modified peptides, pep13, pep5A and pep14A were investigated for their interaction with hIgM/ hIgG1 to elucidate the binding mechanism of pep14. Cysteine in pep14 was identified to be crucial for inducing thiophilic-like interactions between the ligand and hIgM while glutamic acid had a significant role in attributing specificity to the ligand to interact with hIgM. Circular dichroism studies suggested the absence of native β-hairpin structure in pep14. The ligand exhibited a more flexible conformation consisting mainly of a mixture of coil and turn. [Copyright &y& Elsevier]

Abstract: We had previously described the design of a human polymeric immunoglobulin receptor mimetic peptidic ligand, pep14 (CITLISSEGYVSSK), that bound human IgM (hIgM) selectively and exhibited negligible affinity for other Ig proteins. In the present study, pep14 was investigated for its efficacy as an affinity ligand for IgM purification. Binding buffers at varying pH and ionic strengths were investigated. 10mM HEPES containing 150mM NaCl at pH 7.4 was found to be the optimum binding buffer. Equilibrium studies on pep14-immobilized-silica microspheres indicated a binding capacity of 5.9mg hIgM/g of silica–amine microspheres and an association constant of 3.2×106 M−1. Exposure of pep14 to a synthetic mixture comprising hIgM, hIgG1 and bovine serum albumin had no adverse effect on the binding capacity and specificity of pep14. Experimental results highlighted the uniqueness of pep14 in capturing hIgM even at concentrations as low as 10μg/mL. Surface plasmon resonance based studies suggested that while pep14 exhibited affinity to human and rabbit IgM, pep14 failed to interact with mouse IgM. pep14 was resistant to column-sanitizing agents including 20% ethanol and 70% ethanol, although exposure to 100mM and 500mM sodium hydroxide resulted in hydrolysis. [Copyright &y& Elsevier]

Abstract: The polymeric immunoglobulin receptor (pIgR) plays a pivotal role in mucosal immune protection by transporting secretory immunoglobulins to mucosal epithelia, and protecting them from proteolytic degradation. It has been reported that a homolog of the pIgR has a similar role in teleost fish. Considering the role pIgR has in mucosal defenses, this study was initiated to characterize a possible pIgR homolog in Atlantic salmon (Salmo salar) and its relatedness to pIgR of other vertebrates and similar molecules. Two pIgR-like cDNAs and genes of Atlantic salmon (Salsal pIgR and Salsal pIgRL) were cloned and analyzed. In addition, we gathered sequence information of CMRF35-like molecules (CLM) 1, 7, and 8 (designated as CD300 in humans) and made a comparative evaluation to that of the Salsal pIgR and Salsal pIgRL polypeptides. Salsal pIgR and Salsal pIgRL, like pIgR in other teleosts, are composed of two IG V domains, a connecting, a transmembrane, and a cytoplasmic region. The same holds true for Atlantic salmon CLM1 and CLM7, except that they possess putative immunoreceptor tyrosine-based inhibitory motifs (ITIM) in their cytoplasmic tails. The abundance of Salsal pIgR transcript is significantly higher than Salsal pIgRL and CLM in the skin, while Salsal pIgRL transcripts were abundant in the gills, depicting their possible tissue-specific role in mucosal immunity. To further highlight the roles of these molecules in cutaneous mucosal defence, we compared their transcriptional changes in salmon skin and spleen infected with the ectoparasite Lepeophtheirus salmonis which targets skin and mucus of salmonid fish (sampled 3, 14 and 28 days post infection (dpi)). Salsal pIgR and Salsal pIgRL transcripts significantly increased after 14dpi in skin and spleen. CLM1 was up-regulated in skin and down-regulated in spleen, possibly indicating that CLM1 expressing cells had migrated to the target site. Homology modeling using human pIgR domain 1 (PDB 1xed) identified structurally equivalent residues on both Salsal pIgR and Salsal pIgRL, and the same domain disulphide bridge topology. Cysteines 42 and 50 (IMGT numbering) are 7 residues apart in all V domains of Salsal pIgR, Salsal pIgRL, and mammalian [D1]. [Copyright &y& Elsevier]

Abstract: Proteoglycan (P1) purified from Phellinus linteus has been reported to have anti-disease activities. The objectives of our research were to determine the anti-tumor effect and possible mechanisms of P1 on human cancer cells. Cell inhibition assay showed that P1 has an antiproliferative effect on HepG2, HT-29, NCI-H 460 and MCF-7 human colon cancer cells, especially it was very effective in inhibiting HT-29 cells. When HT-29-bearing mice were treated with P1(100mg/kg), there was relative increase in spleen and thymus weights, the plasmatic pIgR and IgA levels were significantly increased, also there was a notable decrease in plasmatic PGE2, Reg IV, EGFR and Akt concentrations measured by ELISA. RT-PCR analysis suggested that P1-induced HT-29 apoptosis appeared to be associated with a decrease in the levels of expression of Reg IV and EGFR. These results suggest that P1 might have two potential roles in treating cancer; it acts as an immunopotentiator partly through protecting T cells from escaping PGE2 attack and enhancing the mucosal IgA response, and as a direct inhibitor by disrupting the Reg IV/EGFR/Akt signaling pathway. [Copyright &y& Elsevier]

Abstract: The abomasal nematode Ostertagia ostertagi is a major causal agent contributing to production inefficiencies in the cattle industry in temperate regions of the world. Protective immunity to infections develops very slowly and resistance to reinfection manifests only after prolonged exposure. Mechanisms underlying the development of protective immunity remain largely unexplored. Immune animals, which have significantly reduced worm burdens, were developed after multiple drug-attenuated experimental infections and were compared to a primary infected group and their respective uninfected controls. In this study, transcriptomic analysis identified three signaling pathways significantly impacted during both primary and repeat infections, the complement system, leukocyte extravasation and acute phase responses. Increased mRNA levels of complement components C3, factor B (CFB) and factor I (CFI) in the abomasal mucosa of the infected cattle were confirmed using quantitative PCR while Western blot analysis established the presence of elevated levels of activated C3 proteins in the mucosa. One of the initiators of local complement activation could be related to secretory IgA and IgM because infections significantly up-regulated expression of J chain (IGJ), as well as polymeric Ig receptor (PIGR) and an IgM-specific receptor (FAIM3), suggesting sustained increases in both synthesis and transepithelial transport of IgA and IgM during the infection. The elevated levels of pro-inflammatory cytokines, such as IL-4 and IL-1β, during infection may be involved in gene regulation of complement components. Our results suggest enhanced tissue repair and mucin secretion in immune animals may also contribute to protective immunity. These results are the first evidence that local complement activation may be involved in the development of long-term protective immunity and provide a novel mechanistic insight into resistance against O. ostertagi in cattle. [ABSTRACT FROM AUTHOR]

Abstract: 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. [Copyright &y& Elsevier]

Abstract: Background: Secretory immunoglobulin A (sIgA) is the principle immune defense against bacteria and other pathogens in the gut and other mucosal surfaces. sIgA is produced locally by plasma cells and transported (transcytosis) across epithelial cells into luminal secretions. sIgA production and transcytosis are governed by multiple signals, which may be affected by T cells. The purpose of this study was to elucidate the role of the Th1-type cytokine, interferon gamma (IFN-γ), and the Th2-type cytokine interleukin-4 (IL-4) on IgA transcytosis across intestinal epithelial cells in vitro. Methods: HT-29 cell monolayers were established in a 2-chamber cell culture system. Cell monolayers were exposed to either IFN-γ, IL-4, or both on the apical or basal side of the culture system systems for 48 hours. Dimeric IgA then was added to the basolateral chamber and transcytosis into the apical chamber was quantified by enzyme-linked immunosorbent assay at timed intervals. IgA transcytosis across HT-29 cells without prior addition of either cytokine served as a control. Results: Poly Ig–receptor up-regulation was shown by both IFN and IL-4. A combination of these 2 cytokines caused the greatest increase in receptor expression. A total of 3.7% of HT-29 cells expressed the polymeric Ig receptor in the control group. This increased to 23.7% in IL-4–treated cells, 66.3% in IFN-γ–treated cells, and 71.5% of cells treated with both cytokines. These findings re-show previously published findings. The effects of IFN-γ added to the basal chamber of HT-29 cell cultures shows a 7- to 10-fold increase in sIgA transcytosis at the 1-, 3-, and 12-hour time intervals. The effect of the addition of IL-4 to the basal chamber was similar to the results noted with IFN-γ alone. The greatest IgA transcytosis was noted when IFN-γ and IL-4 both were added to the basal chamber. However, these same results did not occur with cytokine exposure to the apical side of cell monolayers. Conclusions: IFN-γ and IL-4 increased IgA transport across HT-29 cells in a polar fashion. An increased effect on IgA transport was noted with the combination of IFN-γ and IL-4. Delivery of T-cell cytokines to mucosal surfaces may support local antibody defense of mucosal surfaces and prevent infection. [Copyright &y& Elsevier]

Abstract: Immunoglobulin A (IgA) is transported into saliva by salivary cells expressing the polymeric immunoglobulin receptor (pIgR). In rat salivary glands, autonomic nerves stimulate this process. To examine how nerves affect pIgR-mediated IgA secretion, the chorda-lingual nerve was sectioned. One week after preganglionic parasympathectomy, both the stimulated and unstimulated rates of salivary IgA secretion were reduced, despite similar glandular amounts of IgA. Biochemical analysis of cells from parasympathectomised and control glands indicated reduced membrane expression of pIgR. It appears the removal of long-term parasympathetic input has affected the routing of pIgR within salivary cells and reduced the SIgA transport into saliva. [Copyright &y& Elsevier]

The polymeric immunoglobulin receptor (pIgR) is one of the most vital components of mucosal immunity that plays a pivotal role in mediating transcytosis of polymeric immunoglobulin (pIg) on epithelial surfaces for protection against invading pathogens. Herein, we cloned the full-length cDNA of Pelodiscus sinensis pIgR, designated as P. sinensis pIgR, made of an open reading frame (ORF) of 1848 bp, molecular weight of 68.2 kDa and estimated isoelectric point of 7.00. The deduced P. sinensis pIgR sequence had a leader peptide, extracellular region containing four immunoglobulin-like domains (Ig like domains), transmembrane and intracellular regions comparable with other vertebrates. P. sinensis pIgR contained four Ig like domains that corresponded with mammalian D1, D3, D4 and D5 similar with reptile and avian Ig like domains. It had 40 potential phosphorylation sites, four putative N -glycosylation sites and several motifs resembling mammalian pIgR motifs. Phylogenetic analysis showed a close relationship between P. sinensis pIgR with avian and reptile pIgRs. P. sinensis pIgR basal levels were higher in the esophagus, small intestine and intestinnum crissum than in other organs of health turtles. Intragastric delivery of LPS and Aeromonas sobria led to significant upregulation of P. sinensis pIgR in tissues of the gastrointestinal tract. A polyclonal anti- P. sinensis pIgR antibody produced in rabbit reacted with the recombinant P. sinensis pIgR protein expressed in Escherichia coli in Western blot. These studies demonstrate the existence and immune response of P. sinensis pIgR to stimulation in mucosal organs in Chinese soft-shelled turtles. • Pelodiscus sinensis polymeric immunoglobulin receptor (pIgR) is like other vertebrate pIgRs. • P. sinensis has four immunoglublin like domains. • P. sinensis pIgR basal levels are highest in the gastrointestinal tract than other organs. • P. sinensis pIgR is upregulated by LPS and Aeromonas sobria stimulation. [ABSTRACT FROM AUTHOR]

Polymeric immunoglobulin receptor (pIgR) is a protein that transports Immunoglobulins (Igs) from epithelial cells into the external secretion system of the animal. In the present study, we characterized the partial pIgR gene from Labeo rohita and analyzed its expression in response to the pDNA (pGPD-IFN) vaccine, and studied its correlation with the expression of IgM, in the mucosal-associated lymphoid tissues (MALT). The significant (p < 0.05) up-regulation of pIgR mRNA was observed in the mucosal tissues of vaccinated fish by qRT-PCR. The highest expression of the gene was detected in gill tissue at 15 days post-vaccination (dpv) followed by skin and gut at 30 dpv. The expression of IgM also showed a similar pattern and indicated a direct correlation of pIgR expression in all the tested tissues. The protective immune response of the DNA vaccine was measured as the relative percentage of survival (RPS) by challenging vaccinated fish with live Edwardsiella tarda (1 × 105 CFU/Fish). The result showed a significantly high relative percentage survival (RPS) in the vaccinated group (47.05%) compared to the control group. Many factors contributing to the immune response of the vaccine. One of the most critical aspects is the rise in IgM level in local tissues. In other higher vertebrates, pIgR is reported to act as the transporter of IgM. The positive correlation in the expression of pIgR and IgM observed in the present study demonstrated the possible role of pIgR as the transporter of IgM in L. rohita. The study suggests the possibility of the secretory nature of IgM in fish. • Characterization of polymeric immunoglobulin receptor gene (pIgR) in L. rohita • mRNA expression of pIgR and IgM gene in mucosal associated lymphoid tissues in response to DNA vaccine and E. tarda infection • Vaccine efficacy study against E. tarda infection in terms of relative percentage survival (RPS). [ABSTRACT FROM AUTHOR]