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Background: Chronic Hepatitis B (CHB) is a leading cause of liver fibrosis and cirrhosis worldwide. The early detection of liver fibrosis remains challenging due to the lack of specific symptoms and noninvasive biomarkers with high sensitivity. The polymeric immunoglobulin receptor (PIGR) has recently emerged as a potential biomarker for liver fibrosis. This study aims to evaluate the utility of PIGR in CHB patients as a biomarker for liver fibrosis. Methods: This retrospective study analyzed 150 CHB patients from 2018 to 2023. Based on liver biopsy results, 34 patients were classified as having liver fibrosis, while 116 were categorized as non-fibrosis. Clinical data were compared to assess the relationship between PIGR expression levels and serum fibrosis indices. Logistic regression was performed to identify factors influencing liver fibrosis, and the predictive value of PIGR was evaluated using a receiver operating characteristic (ROC) curve. Results: Significant differences were observed in collagen type IV (CIV), procollagen type III N-terminal peptide (PCIIINP), and hyaluronic acid (HA) levels between the fibrosis and non-fibrosis groups (P < 0.05). PIGR levels were significantly higher in the fibrosis group (P < 0.05) and positively correlated with HA, laminin (LN), PCIII, and CIV levels (P < 0.05). Logistic regression identified HA, LN, PCIIINP, and CIV as risk factors, with PIGR being an independent predictor (P < 0.05). At a cutoff value of 0.35, PIGR showed an area under the curve (AUC) of 0.839, with 81.90% sensitivity, 79.41% specificity, and a Youden's index of 0.613. PIGR also provided a higher net benefit than APRI. Conclusion: PIGR levels are significantly elevated in CHB-related liver fibrosis and correlate closely with established fibrosis markers. As an independent predictor, PIGR demonstrates high diagnostic accuracy and holds promise as a non-invasive biomarker for detecting liver fibrosis in CHB patients, with significant potential for clinical application. [ABSTRACT FROM AUTHOR]

In order to investigate the effect of mannan oligosaccharides on the intestinal barrier of broilers, a total of 540 1-day-old healthy white-feathered broilers were selected for the experiment. Broilers were raised to 29 days old and randomly divided into three groups based on the principle of similar body weight with half male and half female: model control group, MOS health group, and ciprofloxacin lactate health group. Each group had 6 replicates, with 30 replicates each, and the experimental period was 10 d. At the age of 30 d, broilers in the model control group, MOS health group, and ciprofloxacin lactate health group were injected into the pectoral muscle with 8.37×109 CFU/mL Escherichia coli bacterial solution was used for disinfection. Afterwards, drinking water, 0.05 g/L MOS solution, and 60 mg/L ciprofloxacin lactate were administered separately for 10 days. Weigh the broilers after all experiments are completed, blood was collected for serum biochemical indicators, organ indices were measured from various segments of the small intestine, and the expression of key genes in the intestinal barrier of broilers was detected by RT qPCR. The results showed that the body weight and organ index of broilers showed no significant differences among the model control group, MOS health group, and ciprofloxacin lactate health group (P>0.05). Compared with the model control group and the ciprofloxacin lactate health group, the MOS health group broiler serum γ-GGT content significantly increased (P<0.05). Compared with the model control group, the MOS health group showed an increase in PIgR in the duodenum of broiler chickens (P<0.05); compared with the ciprofloxacin lactate health group, the MOS health group showed an increasing trend in the expression of PIgR in the jejunum of broiler chickens (P=0.077). In summary, adding MOS to drinking water increases serum levels γ-GGT upregulates the expression of PIgR in the immune barrier of the duodenum and jejunum of broiler chickens, enhances the intestinal immune barrier of broiler chickens, maintains intestinal health, and thereby improves the immune capacity of the body, with a more significant effect compared to ciprofloxacin lactate. [ABSTRACT FROM AUTHOR]

SARS-CoV-2 is a highly pathogenic respiratory virus that successfully initiates and establishes its infection at the respiratory mucosa. However, little is known about how SARS-CoV-2 antagonizes the host's mucosal immunity. Recent findings have shown a marked reduction in the expression of the polymeric Ig receptor (pIgR) in COVID-19 patients. This receptor maintains mucosal homeostasis by transporting the dimeric IgA (dIgA) and pentameric IgM (pIgM) across mucosal epithelial cells to neutralize the invading respiratory pathogens. By studying the interaction between pIgR and SARS-CoV-2 proteins, we discovered that the viral accessory protein Open Reading Frame 8 (ORF8) potently downregulates pIgR expression and that this downregulation activity of ORF8 correlates with its ability to interact with pIgR. Importantly, the ORF8-mediated downregulation of pIgR diminishes the binding of dIgA or pIgM, and the ORF8 proteins of the variants of concern of SARS-CoV-2 preserve the function of downregulating pIgR, indicating the importance of this conserved activity of ORF8 in SARS-CoV-2 pathogenesis. We further observed that the secreted ORF8 binds to cell surface pIgR, but that this interaction does not trigger the cellular internalization of ORF8, which requires the binding of dIgA to pIgR. These findings suggest the role of ORF8 in SARS-CoV-2 mucosal immune evasion. [ABSTRACT FROM AUTHOR]

Acinetobacter species encode for extracellularly secreted Biofilm-associated protein (Bap), a multi-domain protein with variable molecular weights reaching several hundred kilodaltons. Bap is crucial for the development of multi-dimensional structures of mature biofilms. In our investigation, we analyzed 7338 sequences of A. baumannii from the NCBI database and found that Bap or Bap-like protein (BLP) was present in 6422 (87.52%) isolates. Further classification revealed that 12.12% carried Type-1 Bap, 68.44% had Type-2, 6.91% had Type-3, 0.05% had Type-6 or SDF-Type, and 12.51% lacked Bap or BLP. The majority of isolates with Type-1, Type-2, and Type-3 Bap belonged to ST1, ST2, and ST25, respectively. Phylogenetic analysis suggested that Type-1 Bap is the most ancient, while Type-3 and SDF-Type have evolved recently. Studying the interaction of predicted Bap structures with human CEACAM-1 and PIgR showed that Bap with its BIg13 and BIg6 domains interact with the N-terminal domain of CEACAM-1, involving Arg43 and Glu40, involved in CEACAM-1 dimerization. Also, we found that recently evolved Type-3 and SDF-Type Bap showed greater interaction with CEACAM-1 and PIgR. It can be asserted that the evolution of Bap has conferred enhanced virulence characteristics to A. baumannii with increased interaction with CEACAM-1 and PIgR. Using in silico approaches, this study explores the evolutionary, physicochemical, and structural features of A. baumannii Bap and unravels its crucial role in mediating interaction with human CEACAM-1 and PIgR through detailed structure modelling. These findings advance our understanding of A. baumannii Bap and highlight its role in pathogenesis. [ABSTRACT FROM AUTHOR]

BackgroundThe genetic factors play important roles on the pathogenesis of inflammatory bowel disease (IBD). EpCAM is highly expressed in the intestinal epithelium. It is still unclear if the decrease or somatic mutation of EpCAM could cause IBD.MethodsThe WT and EpCAM+/− mice were administrated with DSS intermittently for nearly 8 weeks. The colon, liver and feces were harvested to check the morphological and histological changes, the expression of inflammatory genes and the gut microbiota via H&E staining, immunofluorescence, qPCR, western blot and 16S rDNA sequence assays.ResultsThe DSS administration induced more serious inflammation in the colon of EpCAM+/− mice than WT mice. Compared to DSS-induced WT mice, the transcriptional levels of IL-6, F4/80, Ly6g, Ly6d and Igha were significantly higher in the colon of DSS-induced EpCAM+/− mice. The protein levels of MMP7 and MMP8 and the activation of JNK, ERK1/2 and p38 were significantly increased in the colon of DSS-induced EpCAM+/− mice. The protein levels of CLDN1, CLDN2, CLDN3, CLDN7, OCLD, ZO-1 and pIgR were significantly decreased in the colon of DSS-induced EpCAM+/− mice. The serum concentration of LPS was significantly higher in the DSS-induced EpCAM+/− mice which caused the acute inflammation in the liver of them. The expression of Pigr was significantly reduced in the liver of DSS-induced EpCAM+/− mice. The ratio of Firmicutes/Bacteroidetes at the phylum level was higher in the gut microbiota of EpCAM+/− mice than WT mice.ConclusionIn conclusion, the heterozygous mutation of EpCAM increased the susceptibility to colitis, gut microbiota dysbiosis and liver injury.

Cows produce saliva in very large quantities to lubricate and facilitate food processing. Estimates indicate an amount of 50–150 L per day. Human saliva has previously been found to contain numerous antibacterial components, such as lysozyme, histatins, members of the S-100 family and lactoferrin, to limit pathogen colonization. Cows depend on a complex microbial community in their digestive system for food digestion. Our aim here was to analyze how this would influence the content of their saliva. We therefore sampled saliva from five humans and both nose secretions and saliva from six cows and separated the saliva on SDS-PAGE gradient gels and analyzed the major protein bands with LC-MS/MS. The cow saliva was found to be dominated by a few major proteins only, carbonic anhydrase 6, a pH-stabilizing enzyme and the short palate, lung and nasal epithelium carcinoma-associated protein 2A (SPLUNC2A), also named bovine salivary protein 30 kDa (BSP30) or BPIFA2B. This latter protein has been proposed to play a role in local antibacterial response by binding bacterial lipopolysaccharides (LPSs) and inhibiting bacterial growth but may instead, according to more recent data, primarily have surfactant activity. Numerous peptide fragments of mucin-5B were also detected in different regions of the gel in the MS analysis. Interestingly, no major band on gel was detected representing any of the antibacterial proteins, indicating that cows may produce them at very low levels that do not harm the microbial flora of their digestive system. The nose secretions of the cows primarily contained the odorant protein, a protein thought to be involved in enhancing the sense of smell of the olfactory receptors and the possibility of quickly sensing potential poisonous food components. High levels of secretory IgA were also found in one sample of cow mouth drippings, indicating a strong upregulation during an infection. The human saliva was more complex, containing secretory IgA, amylase, carbonic anhydrase 6, lysozyme, histatins and a number of other less abundant proteins, indicating a major difference to the saliva of cows that show very low levels of antibacterial components, most likely to not harm the microbial flora of the rumen. [ABSTRACT FROM AUTHOR]

Simple Summary: IgT is a specific type of antibody found in teleost fish that is crucial for protecting mucosal surfaces. However, studying IgT is challenging due to the limited available cell markers. Here, we investigated the distribution of IgT mRNA+ cells and pIgR mRNA+ cells (essential for Ig transport over cell membranes) in Atlantic salmon (Salmo salar) intestines. Using in situ hybridization, we examined two different sizes (developmental stages) of the fish and compared the hybridization-positive cell distribution between vaccinated and unvaccinated fish. Our findings revealed that IgT mRNA+ cells were mainly located beneath the intestinal mucosa, specifically in the lamina propria, while pIgR mRNA+ cells were found in both the lamina propria and mucosa. Additionally, vaccinated fish exhibited abdominal adhesions with tissue containing IgT and pIgR mRNA+ cells. We observed regional variations in the distribution of IgT mRNA+ cells within the salmon intestine that were unaffected by intraperitoneal vaccination but sensitive to fish age. This study provides new insights into the distribution and dynamics of IgT and pIgR mRNA+ cells, advancing our understanding of the spatial mucosal immune system and its implications for teleosts, with potential applications in aquaculture. The newly discovered IgT+ B cell is thought to play a dominant role in mucosal immunity, but limited studies have examined its distribution in fish species, hindering our understanding of its function. This study investigated IgT and poly Ig receptor (pIgR) mRNA+ cell distribution in Atlantic salmon (Salmo salar) gut using RNAscope in situ hybridization (ISH) and assessed the effects of vaccination. The pyloric caeca, mid-intestine (first and second parts), and posterior segment in two weight stages (Group 1: avg. 153 g, Group 2: avg. 1717 g) were examined in both vaccinated and unvaccinated fish. ISH revealed more IgT mRNA+ cells in the second part of the midgut compared to other intestinal segments, as well as a higher number of positive cells in Group 2 (older fish). In line with previous findings, intraperitoneal vaccination had no significant impact on the number of IgT+ transcripts. IgT mRNA+ cells were found mostly in the lamina propria and near capillaries, while pIgR was registered in both the lamina propria and mucosa. Interestingly, vaccinated fish presented adhesions and granulomatous tissue in the peritoneum, with both IgT and pIgR mRNA+ cells. Taken together, these results suggest that the distribution of IgT mRNA+ cells in the intestine of Atlantic salmon is region-specific and is not affected by intraperitoneal vaccination but varies with fish age. [ABSTRACT FROM AUTHOR]

HU protein, a member of the nucleoid-associated group of proteins, is an important transcription factor in bacteria, including in the dangerous human pathogen Francisella tularensis. Generally, HU protein acts as a DNA sequence non-specific binding protein and it is responsible for winding of the DNA chain that leads to the separation of transcription units. Here, we identified potential HU protein binding sites using the ChIP-seq method and two possible binding motifs in F. tularensis subsp. holarctica FSC200 depending upon growth conditions. We also confirmed that FSC200 HU protein is able to introduce negative supercoiling of DNA in the presence of topoisomerase I. Next, we showed interaction of the HU protein with a DNA region upstream of the pigR gene and inside the clpB gene, suggesting possible regulation of PigR and ClpB expression. Moreover, we showed that arginine 58 and partially arginine 61 are important for HU protein’s DNA binding capacity, negative supercoiling induction by HU, and the length and winding of FSC200 chromosomal DNA. Finally, in order to verify biological function of the HU protein, we demonstrated that mutations in arginine 58, arginine 61, and serine 74 of the HU protein decrease virulence of FSC200 both in vitro and in vivo and that immunization using these mutant strains is able to protect as many as 100% of mice against wild-type challenge. Taken together, our findings deepen knowledge about the role of the HU protein in tularaemia pathogenesis and suggest that HU protein should be addressed in the context of tularaemia vaccine development.

Abstract Background Identification of COPD patients with a rapid decline in FEV1 is of particular interest for prognostic and therapeutic reasons. Objective To determine the expression of markers of inflammation in COPD patients with rapid functional decline in comparison to slow or no decliners. Methods In COPD patients monitored for at least 3 years (mean ± SD: 5.8 ± 3 years) for lung functional decline, the expression and localization of inflammatory markers was measured in bronchial biopsies of patients with no lung functional decline (FEV1% + 30 ± 43 ml/year, n = 21), slow (FEV1% ml/year, − 40 ± 19, n = 14) and rapid decline (FEV1% ml/year, − 112 ± 53, n = 15) using immunohistochemistry. ELISA test was used for polymeric immunoglobulin receptor (pIgR) quantitation “in vitro”. Results The expression of secretory IgA was significantly reduced in bronchial epithelium (p = 0.011) and plasma cell numbers was significantly reduced in the bronchial lamina propria (p = 0.017) of rapid decliners compared to no decliners. Bronchial inflammatory cell infiltration, CD4, CD8, CD68, CD20, NK, neutrophils, eosinophils, mast cells, pIgR, was not changed in epithelium and lamina propria of rapid decliners compared to other groups. Plasma cells/mm2 correlated positively with scored total IgA in lamina propria of all patients. “In vitro” stimulation of 16HBE cells with LPS (10 μg/ml) and IL-8 (10 ng/ml) induced a significant increase while H2O2 (100 μM) significantly decreased pIgR epithelial expression. Conclusion These data show an impaired humoral immune response in rapid decliners with COPD, marked by reduced epithelial secretory IgA and plasma cell numbers in the bronchial lamina propria. These findings may help in the prognostic stratification and treatment of COPD.

Cholecystokinin 8 (CCK8) is an entero-octapeptide that participates in crosstalk with components of intestinal immunity via the CCK receptor (CCKR), but its role in modulation of the IgA response is not fully known under physiological conditions. Male eight-week-old BALB/c mice each were intraperitoneally injected once during 7 days with CCK8, devazapide (CCKR1 antagonist), L365,260 (CCKR2 antagonist) or vehicle (sham group). In intestinal lavages, total and secretory IgA (SIgA) were determined by ELISA; in lamina propria, IgA+ B lymphocytes and IgA+ plasma cells were analyzed by flow cytometry; mRNA levels of polymeric immunoglobulin receptor (pIgR) in epithelial cells and α chain, interleukins (ILs) in lamina propria cells were assessed by qRTPCR. Regarding the sham conditions, IgA+ plasma-cell percentage and IL-2, IL-5, IL-10 and transforming growth factor-β (TGF-β) mRNA levels were either increased by CCK8 or decreased by both CCKR antagonists. For IgA/SIgA responses, IL-4/IL-6 mRNA levels were decreased by all drugs and pIgR mRNA was increased by CCK8 and reduced by L365,260. IgA+ B cell percentage and α chain mRNA levels were elicited by CCK8 and L365,260. Data suggested a presumable differential role of CCK/CCKR on the IgA-response; outcome of L365,260 on the elicitation of IgA+ B cells and α chain mRNA needs further examination.

IgA deficiency is the commonest immunodeficiency affecting up to 1 in 700 individuals. The effects of IgA deficiency are difficult to see in many individuals, are mild in many fewer and severe in fewer still. While monovalent IgA is found in serum, dimeric IgA is secreted through mucosal surfaces where it helps to maintain epithelial homeostasis. Studies with knockout mice have taught us that there are subtle inflammatory consequences of removing secretory IgA (sIgA), and the best explanation for these changes can be related by the loss of the 'excretory' immune system. The excretion of antigens is a logical process in regulating the immune system, given the long half-life of complement fixing antibodies. But the function of IgA as an immune or inflammation regulator may go beyond antigen removal. [ABSTRACT FROM AUTHOR]

IgA deficiency is the commonest immunodeficiency affecting up to 1 in 700 individuals. The effects of IgA deficiency are difficult to see in many individuals, are mild in many fewer and severe in fewer still. While monovalent IgA is found in serum, dimeric IgA is secreted through mucosal surfaces where it helps to maintain epithelial homeostasis. Studies with knockout mice have taught us that there are subtle inflammatory consequences of removing secretory IgA (sIgA), and the best explanation for these changes can be related by the loss of the ‘excretory’ immune system. The excretion of antigens is a logical process in regulating the immune system, given the long half-life of complement fixing antibodies. But the function of IgA as an immune or inflammation regulator may go beyond antigen removal.

Polymeric immunoglobulin receptor (pIgR) is an important immune factor in the mucosal immune system of fish, which plays a key role in mediating the secretion and transport of immunoglobulin into mucus. In this study, the full-length cDNA sequence of Megalobrama amblycephala pIgR gene was firstly cloned and the immune response to Aeromonas hydrophila was detected. After being challenged by Aeromonas hydrophila at 3 d, significantly pathological features were observed in intestine, head kidney, spleen, liver and gill of Megalobrama amblycephala. The content of lysozyme (Lys) and the activities of acid phosphatase (ACP) and alkaline phosphatase (AKP) increased significantly at 1 d and reached the peak at 3 d, and the activities of total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-PX) and catalase (CAT) in serum reached the peak at 5 d and 7 d after infection, respectively. The expression level of IL-1β gene reached the peak at 3 d in intestine, 5 d in gill and spleen, 7 d in head kidney and liver of Megalobrama amblycephala after infected by Aeromonas hydrophila , respectively. The TNF-α gene expression reached the peak at 3 d in intestine and gill, 5 d in head kidney and spleen, 7 d in liver after infection, respectively. The experimental results showed that the infection of Aeromonas hydrophila caused the pathological changes of immune-related tissues and triggered the inflammation responses. The full-length cDNA sequence of Megalobrama amblycephala pIgR was 1828 bp, and its open reading frame (ORF) was 1023 bp, encoding 340 amino acids. The pIgR of Megalobrama amblycephala has a signal peptide sequence, followed by extracellular region, transmembrane region and intracellular region. The extracellular region includes two Ig-like domains (ILDs), and its tertiary structure is twisted "L". The phylogenetic tree was constructed using the adjacency method, and the pIgR genes of Megalobrama amblycephala and cyprinidae fish were clustered into a single branch. Quantitative real-time PCR (qRT-PCR) was used to detect the expression of pIgR gene in different tissues of Megalobrama amblycephala. The expression level of pIgR gene was the highest in liver, followed by intestine, head kidney, skin, middle kidney and spleen, lower in heart, gill and brain, and the lowest in muscle. After being infected by Aeromonas hydrophila , the expression level of Megalobrama amblycephala pIgR gene in intestine, head kidney, spleen, liver and gill showed a trend of increasing first and then decreasing within 28 d. The pIgR gene expression reached the peak in mucosal immune-related tissues (gill and intestine) was earlier than that in systemic immune-related tissues (head kidney and spleen), and the relative expression level of pIgR gene at peak in intestine (12.3 fold) was higher than that in head kidney (3.73 fold) and spleen (7.84 fold). These results suggested that Megalobrama amblycephala pIgR might play an important role in the mucosal immune system to against Aeromonas hydrophila infection. • The full-length cDNA of M. amblycephala pIgR was firstly cloned and characterized. • The M. amblycephala pIgR was highly expressed in liver, intestine and head kidney. • Significantly morphological changes in intestine and gill after A. hydrophila infection. • The expression of pIgR was significantly increased in intestine, gill, head kidney, spleen and liver after challenge. • The antioxidant and immune-related enzyme activities were significantly increased in serum after challenge. [ABSTRACT FROM AUTHOR]

Background: Identification of COPD patients with a rapid decline in FEV1 is of particular interest for prognostic and therapeutic reasons.Objective: To determine the expression of markers of inflammation in COPD patients with rapid functional decline in comparison to slow or no decliners.Methods: In COPD patients monitored for at least 3 years (mean ± SD: 5.8 ± 3 years) for lung functional decline, the expression and localization of inflammatory markers was measured in bronchial biopsies of patients with no lung functional decline (FEV1% + 30 ± 43 ml/year, n = 21), slow (FEV1% ml/year, - 40 ± 19, n = 14) and rapid decline (FEV1% ml/year, - 112 ± 53, n = 15) using immunohistochemistry. ELISA test was used for polymeric immunoglobulin receptor (pIgR) quantitation "in vitro".Results: The expression of secretory IgA was significantly reduced in bronchial epithelium (p = 0.011) and plasma cell numbers was significantly reduced in the bronchial lamina propria (p = 0.017) of rapid decliners compared to no decliners. Bronchial inflammatory cell infiltration, CD4, CD8, CD68, CD20, NK, neutrophils, eosinophils, mast cells, pIgR, was not changed in epithelium and lamina propria of rapid decliners compared to other groups. Plasma cells/mm2 correlated positively with scored total IgA in lamina propria of all patients. "In vitro" stimulation of 16HBE cells with LPS (10 μg/ml) and IL-8 (10 ng/ml) induced a significant increase while H2O2 (100 μM) significantly decreased pIgR epithelial expression.Conclusion: These data show an impaired humoral immune response in rapid decliners with COPD, marked by reduced epithelial secretory IgA and plasma cell numbers in the bronchial lamina propria. These findings may help in the prognostic stratification and treatment of COPD. [ABSTRACT FROM AUTHOR]

The IgM and IgT classes were previously identified and characterized in the Antarctic teleost Trematomus bernacchii, a species belonging to the Perciform suborder Notothenoidei. Herein, we characterized the gene encoding the polymeric immunoglobulin receptor (pIgR) in the same species and compared it to the pIgR of multiple teleost species belonging to five perciform suborders, including 11 Antarctic and 1 non-Antarctic (Cottoperca gobio) notothenioid species, the latter living in the less-cold peri-Antarctic sea. Antarctic pIgR genes displayed particularly long introns marked by sites of transposable elements and transcription factors. Furthermore, analysis of T. bernacchii pIgR cDNA unveiled multiple amino acid substitutions unique to the Antarctic species, all introducing adaptive features, including N-glycosylation sequons. Interestingly, C. gobio shared most features with the other perciforms rather than with the cold-adapted relatives. T. bernacchii pIgR transcripts were predominantly expressed in mucosal tissues, as indicated by q-PCR and in situ hybridization analysis. These results suggest that in cold-adapted species, pIgR preserved its fundamental role in mucosal immune defense, although remarkable gene structure modifications occurred. [ABSTRACT FROM AUTHOR]

Polymeric immunoglobulin receptor (pIgR) is an important member of the immunoglobulin superfamily. It can mediate polarity transportation of polymeric immunoglobulin in mucosal epithelial cells and play an important role in innate and acquired immunity. In this study, the full‐length cDNA sequence of Carassius auratus indigentiaus pIgR gene was firstly cloned and characterized. The complete cDNA sequence of C. auratus indigentiaus pIgR gene is 1399 bp, including a 984 bp open reading frame that encodes 327 amino acids. The pIgR is composed of extracellular, transmembrane and intracellular regions, which are composed of 255, 23 and 49 amino acids, respectively. The extracellular region contains two Ig‐like domains (ILDs), ILD1 consists of 104 amino acids (25–128) and ILD2 consists of 93 amino acids (137–229). The similarity between the pIgR sequences of C. auratus indigentiaus and other fish is 65.95%–97.55%. Analysis of phylogenetic trees shows that pIgR of C. auratus indigentiaus clusters into one branch with the pIgR of Cypriniformes. QRT‐PCR analysis revealed that the pIgR gene expression was highest in liver, followed by intestine, gill, head kidney and skin. After challenge with Aeromonas hydrophila, the relative expressions of pIgR in different tissues first increased and then decreased within 96 h. Moreover, the expression of pIgR gene peaked earlier in mucosal immune tissues (intestines, gill and skin) than in system immune tissues. It suggested that pIgR might play an important role in mucosa‐associated lymphoid tissue to protect the C. auratus indigentiaus against Aeromonas hydrophila infection. [ABSTRACT FROM AUTHOR]

Chronic obstructive pulmonary disease (COPD), asthma and cystic fibrosis (CF) are distinct respiratory diseases that share features such as the obstruction of small airways and disease flare-ups that are called exacerbations and are often caused by infections. Along the airway epithelium, immunoglobulin (Ig) A contributes to first line mucosal protection against inhaled particles and pathogens. Dimeric IgA produced by mucosal plasma cells is transported towards the apical pole of airway epithelial cells by the polymeric Ig receptor (pIgR), where it is released as secretory IgA. Secretory IgA mediates immune exclusion and promotes the clearance of pathogens from the airway surface by inhibiting their adherence to the epithelium. In this review, we summarize the current knowledge regarding alterations of the IgA/pIgR system observed in those major obstructive airway diseases and discuss their implication for disease pathogenesis. [ABSTRACT FROM AUTHOR]

EpCAM deficiency causes congenital tufting enteropathy (CTE) which is considered as one kinds of very early onset inflammatory bowel disease (IBD). However, functions of EpCAM on regulating the immunity of intestines are still unclear. To study the mechanism of EpCAM on maintaining the intestinal immune homeostasis, the intestines of WT and EpCAM-/- mice at E18.5, P0 and P3 stages were collected for morphological, histological and gene expression tests. Serious inflammation was detected in the small intestines of P3 EpCAM-/- mice. Compared to WT mice, genes related to inflammatory factors and immunity cells, including TNFα, IL-1β, IL-6, IL-8rb, MIP2, MCP1, Ly6d and Ly6g, were all significantly upregulated and the expression of intestinal abundance matrix metalloproteinases (MMPs) was also significantly increased in the intestines of EpCAM-/- mice at E18.5, P0 and P3 stages. Signals of p38, ERK1/2 and JNK were hyper-activated in the intestines of EpCAM-/- mice. The expression of pIgR was significantly decreased and the expression and activation of transcriptional factors which promote the expression of pIgR were also reduced in the intestines of EpCAM-/- mice compared to WT controls. In conclusion, EpCAM could maintain the immune homeostasis of intestines via keeping the expression of pIgR in the intestinal epithelium. [ABSTRACT FROM AUTHOR]

EpCAM deficiency causes congenital tufting enteropathy (CTE) which is considered as one kinds of very early onset inflammatory bowel disease (IBD). However, functions of EpCAM on regulating the immunity of intestines are still unclear. To study the mechanism of EpCAM on maintaining the intestinal immune homeostasis, the intestines of WT and EpCAM-/- mice at E18.5, P0 and P3 stages were collected for morphological, histological and gene expression tests. Serious inflammation was detected in the small intestines of P3 EpCAM-/- mice. Compared to WT mice, genes related to inflammatory factors and immunity cells, including TNFα, IL-1β, IL-6, IL-8rb, MIP2, MCP1, Ly6d and Ly6g, were all significantly upregulated and the expression of intestinal abundance matrix metalloproteinases (MMPs) was also significantly increased in the intestines of EpCAM-/- mice at E18.5, P0 and P3 stages. Signals of p38, ERK1/2 and JNK were hyper-activated in the intestines of EpCAM-/- mice. The expression of pIgR was significantly decreased and the expression and activation of transcriptional factors which promote the expression of pIgR were also reduced in the intestines of EpCAM-/- mice compared to WT controls. In conclusion, EpCAM could maintain the immune homeostasis of intestines via keeping the expression of pIgR in the intestinal epithelium.

The polyimmunoglobulin receptor (pIgR) transcytoses J chain‐containing antibodies through mucosal epithelia. In mammals, two cis‐duplicates of PIGR, FCMR, and FCAMR, flank the PIGR gene. A PIGR duplication is first found in amphibians, previously annotated as PIGR2 (herein xlFCAMR), and is expressed by APCs. We demonstrate that xlFcamR is the equivalent of mammalian FcamR. It has been assumed that pIgR is the oldest member of this family, yet our data could not distinguish whether PIGR or FCAMR emerged first; however, FCMR was the last family member to emerge. Interestingly, bony fish "pIgR" is not an orthologue of tetrapod pIgR, and possibly acquired its function via convergent evolution. PIGR/FCAMR/FCMR are members of a larger superfamily, including TREM, CD300, and NKp44, which we name the "double‐disulfide Ig superfamily" (ddIgSF). Domains related to each ddIgSF family were identified in cartilaginous fish (sharks, chimeras) and encoded in a single gene cluster syntenic to the human pIgR locus. Thus, the ddIgSF families date back to the earliest antibody‐based adaptive immunity, but apparently not before. Finally, our data strongly suggest that the J chain arose in evolution only for Ig multimerization. This study provides a framework for further studies of pIgR and the ddIgSF in vertebrates. [ABSTRACT FROM AUTHOR]

We previously reported that exclusively breastfed infants born to mothers with pregestational obesity gain less weight during the first month after birth than those born to mothers of normal pregestational weight. This issue is potentially important since lower weight gain in breastfed infants of obese mothers might increase the risk of developing later obesity. Breast milk quality and quantity, together with breastfeeding practice, possibly influence infants’ feeding behavior, appetite control, and regulation of growth later in life. The issue of whether breast milk protein patterns from obese mothers differ in composition from those of non‐obese mothers remains largely unexplored. Here, we established a breast milk proteomic pattern that discriminates obese mothers and infants with delayed weight gain at 1 month after birth from normal‐weight mothers with infants of the same age and with normal weight gain. Obese mothers were matched to normal‐weight mothers (n = 26; body mass index 33.5 ± 3.2 vs 21.5 ± 1.5 kg·m−2). The mean weight gain of infants in the obese group at 1 month after birth was 430.8 g lower than that of the infants in the control group. Analysis of the breast milk delipidized fraction by surface‐enhanced laser desorption/ionization on CM10 and Q10 arrays was followed by MS‐assisted purification and LC‐MS/MS microsequencing of a selected biomarker. We identified 15 candidate protein biomarkers, seven of which were overexpressed in the obese group and eight in the normal‐weight group. One of the most significant candidate biomarkers, overexpressed in the obese group, was identified as a fragment of the sixth extracellular domain of the polymeric immunoglobulin receptor. Further structural identification of these candidate biomarkers and their validation in clinical assays may facilitate the development of a predictive immunoassay.

The global health crisis and economic tolls of COVID-19 necessitate a panoply of strategies to treat SARS-CoV-2 infection. To date, few treatment options exist, although neutralizing antibodies against the spike glycoprotein have proven to be effective. Because infection is initiated at the mucosa and propagates mainly at this site throughout the course of the disease, blocking the virus at the mucosal milieu should be effective. However, administration of biologics to the mucosa presents a substantial challenge. Here, we describe bifunctional molecules combining single-domain variable regions that bind to the polymeric Ig receptor (pIgR) and to the SARS-CoV-2 spike protein via addition of the ACE2 extracellular domain (ECD). The hypothesis behind this design is that pIgR will transport the molecule from the circulation to the mucosal surface where the ACE ECD would act as a decoy receptor for the nCoV2. The bifunctional molecules bind SARS-Cov-2 spike glycoprotein in vitro and efficiently transcytose across the lung epithelium in human tissue-based analyses. Designs featuring ACE2 tethered to the C-terminus of the Fc do not induce antibody-dependent cytotoxicity against pIgR-expressing cells. These molecules thus represent a potential therapeutic modality for systemic administration of neutralizing anti-SARS-CoV-2 molecules to the mucosa.

Immunoglobulins are glycoproteins which are produced as membrane-bound receptors on B-cells or in a secreted form, known as antibodies. In teleosts, three immunoglobulin isotypes, IgM, IgT, and IgD, are present, each comprising two identical heavy and two identical light polypeptide chains. The basic mechanisms for generation of immunoglobulin diversity are similar in teleosts and higher vertebrates. The B-cell pre-immune repertoire is diversified by VDJ recombination, junctional flexibility, addition of nucleotides, and combinatorial association of light and heavy chains, while the post-immune repertoire undergoes somatic hypermutation during clonal expansion. Typically, the teleost immunoglobulin heavy chain gene complex has a modified translocon arrangement where the Dτ–Jτ–Cτ cluster of IgT is generally located between the variable heavy chain (VH) region and the Dμ/δ–Jμ/δ–Cμ–Cδ gene segments, or within the set of VH gene segments. However, multiple genome duplication and deletion events and loss of some individual genes through evolution has complicated the IgH gene organization. The IgH gene arrangement allows the expression of either IgT or IgM/IgD. Alternative splicing is responsible for the regulation of IgM/IgD expression and the secreted versus transmembrane forms of IgT, IgD, and IgM. The overall structure of IgM and IgT is usually conserved across species, whereas IgD has a large variety of structures. IgM is the main effector molecule in both systemic and mucosal immunity and shows a broad range of concentrations in different teleost species. Although IgM is usually present in higher concentrations under normal conditions, IgT is considered the main mucosal Ig. [ABSTRACT FROM AUTHOR]

The effects of the secondary metabolite biosynthesis on the metabolism and morphology of the Monascus purpureus were investigated in this study. Hypha and septum length became longer after deletion of genes pigR and pksCT in M. purpureus LQ-6 by Agrobacterium tumefaciens-mediated transformation technology, highly branched hyphae, much smaller and freely dispersed mycelial pellets were observed in M. purpureus. Compared with that in the wild-type, the level of intracellular NADH and NADPH was almost constant in M. purpureus ΔpigR at 4 days, but the NADH and NADPH levels decreased by 1.58-fold and 3.71-fold in M. purpureus ΔpksCT. The present study can not only provide a kind of strategy to improve the Monascus pigments production, but also provide theoretical support for the further study of relationship between the secondary metabolites, metabolism and morphological change. [ABSTRACT FROM AUTHOR]

Fish rely on mucosal surfaces as their first defence barrier against pathogens. Maintaining mucosal homeostasis is therefore crucial for their overall well-being, and it is likely that secreted immunoglobulins (sIg) play a pivotal role in sustaining this balance. In mammals, the poly-Ig receptor (pIgR) is an essential component responsible for transporting polymeric Igs across mucosal epithelia. In teleost fish, a counterpart of pIgR has been identified and characterized, exhibiting structural differences and broader mRNA expression patterns compared to mammals. Despite supporting evidence for the binding of Igs to recombinant pIgR proteins, the absence of a joining chain (J-chain) in teleosts challenges the conventional understanding of Ig transport mechanisms. The transport of IgM to the intestine via the hepatobiliary route is observed in vertebrates and has been proposed in a few teleosts. Investigations on the stomachless fish, ballan wrasse, revealed a significant role of the hepatobiliary route and interesting possibilities for alternative IgM transport routes that might include pancreatic tissue. These findings highlight the importance of gaining a thorough understanding of the mechanisms behind Ig transport to the gut in various teleosts. This review aims to gather existing information on pIgR-mediated transport across epithelial cells and immunoglobulin transport pathways to the gut lumen in teleost fish. It provides comparative insights into the hepatobiliary transport of Igs to the gut, emphasizing the current understanding in teleost fish while exploring potential alternative pathways for Ig transport to the gut lumen. Despite significant progress in understanding various aspects, there is still much to uncover, especially concerning the diversity of mechanisms across different teleost species. [Display omitted] • Secreted immunoglobulins (sIg) are crucial for defending mucosal surfaces against pathogens and maintaining homeostasis. • Teleosts have a pIgR counterpart to mammals, yet understanding Ig transport remains challenging. • Exploring IgM transport in teleosts reveals intriguing possibilities, such as pancreatic involvement in ballan wrasse. [ABSTRACT FROM AUTHOR]

Retromer is a multimeric protein complex that mediates intracellular receptor sorting. One of the roles of retromer is to promote transcytosis of the polymeric immunoglobulin receptor (pIgR) and its ligand polymeric immunoglobulin A (pIgA) in polarized epithelial cells. In Madin-Darby Canine Kidney (MDCK) cells, overexpression of Vps35, the retromer subunit key for cargo recognition, restores transcytosis to a pIgR mutant that is normally degraded. Here we show that pIgA transcytosis was not restored in these cells when treated with the specific phosphoinositide 3-kinase (PI3K) inhibitor LY294002. Likewise, the decrease in pIgA transcytosis by wild-type pIgR seen upon PI3K inhibition was not reverted by Vps3S overexpression. PI3K inhibition reduced membrane association of sorting-nexins (SNX) 1 and 2, which constitute the retromer subcomplex involved in membrane deformation, while association of the Vps35-Vps26-Vps29 subcomplex, involved in cargo recognition, remained virtually unaffected. Colocalization between the two retromer subcomplexes was reduced upon the treatment. Whereas the interaction among the subunits of the Vps3S Vps26-Vps29 subcomplex remained unchanged, less Vps35 was found associated with pIgR upon P13K inhibition. In addition, colocalization of internalized pIgA with subunits of both retromer subcomplexes throughout the transcytotic pathway was substantially reduced by LY294002 treatment. These data implicate P13K in controlling retromer's role in pIgR-pIgA transcytosis. (c) 2006 Elsevier Inc. All rights reserved.

We previously reported that exclusively breastfed infants born to mothers with pregestational obesity gain less weight during the first month after birth than those born to mothers of normal pregestational weight. This issue is potentially important since lower weight gain in breastfed infants of obese mothers might increase the risk of developing later obesity. Breast milk quality and quantity, together with breastfeeding practice, possibly influence infants' feeding behavior, appetite control, and regulation of growth later in life. The issue of whether breast milk protein patterns from obese mothers differ in composition from those of non‐obese mothers remains largely unexplored. Here, we established a breast milk proteomic pattern that discriminates obese mothers and infants with delayed weight gain at 1 month after birth from normal‐weight mothers with infants of the same age and with normal weight gain. Obese mothers were matched to normal‐weight mothers (n = 26; body mass index 33.5 ± 3.2 vs 21.5 ± 1.5 kg·m−2). The mean weight gain of infants in the obese group at 1 month after birth was 430.8 g lower than that of the infants in the control group. Analysis of the breast milk delipidized fraction by surface‐enhanced laser desorption/ionization on CM10 and Q10 arrays was followed by MS‐assisted purification and LC‐MS/MS microsequencing of a selected biomarker. We identified 15 candidate protein biomarkers, seven of which were overexpressed in the obese group and eight in the normal‐weight group. One of the most significant candidate biomarkers, overexpressed in the obese group, was identified as a fragment of the sixth extracellular domain of the polymeric immunoglobulin receptor. Further structural identification of these candidate biomarkers and their validation in clinical assays may facilitate the development of a predictive immunoassay. A breast milk proteomic pattern of 15 candidate biomarkers, established by surface‐enhanced laser desorption/ionization, delineates a specific group of obese mothers and infants with delayed weight gain at 1 month after birth. One of these biomarkers was purified and identified by LC‐MS/MS as a 49‐amino‐acid fragment of the sixth extracellular domain of the polymeric immunoglobulin receptor. [ABSTRACT FROM AUTHOR]

Exosomes are small extracellular vesicles (EVs) found in culture supernatants, blood, and breast milk. The size of these nanocomplexes limits the methods of EV analyses. In this study, nitrobenzoxadiazole (NBD), a fluorophore, conjugated endosome-lysosome imager, GIF-2250 and its derivative, GIF-2276, were evaluated for exosome analyses. A correlation was established between GIF-2250 intensity and protein maker levels in bovine milk exosomes. We found that high-temperature sterilization milk may not contain intact exosomes. For precise analysis, we synthesized GIF-2276, which allows for the covalent attachment of NBD to the Lys residue of exosome proteins, and labeled milk exosomes were separated using a gel filtration system. GIF-2276 showed chromatographic peaks of milk exosomes containing >3 ng protein. The area (quantity) and retention time (size) of the exosome peaks were correlated to biological activity (NO synthesis suppression in RAW264.7 murine macrophages). Heat denaturation of purified milk-derived exosomes disrupted these indicators. Proteome analyses revealed GIF-2276-labeled immunomodulators, such as butyrophilin subfamily 1 member A1 and polymeric immunoglobulin receptor. The immunogenicity and quantity of these factors decreased by heat denaturation. When milk exosomes were purified from market-sourced milk we found that raw and low-temperature sterilization milk samples, contained exosomes (none in high-temperature sterilization milk). These results were also supported by transmission electron microscopy analyses. We also found that GIF-2276 could monitor exosome transportation into HEK293 cells. These results suggested that GIF-2250/2276 may be helpful to evaluate milk exosomes. [Display omitted] • GIF-2250 visualizes milk exosomes in response to their quality. • The covalent binder GIF-2276 separates milk exosomes in a gel-filtration system. • GIF-2276 targets the immunomodulators pIgR and BTN1A1 in milk exosomes. • pIgR and BTN1A1 are temperature sensitive factors that are possible quality makers of milk exosomes. [ABSTRACT FROM AUTHOR]

The aim of the present study was to explore the mechanism of agonists in regulating transcriptional level of polymeric immunoglobulin receptor (pIgR) in salivary gland epithelial cells, thus revealing the defense effect of salivary immune on bacteria in the oral cavity. Sixty patients with oral bacterial infection and 70 patients suffering from oral diseases without bacterial infection were selected randomly from patients in Renmin Hospital of Wuhan University from April 2015 to April 2017. Ribonucleic acid (RNA) was extracted from salivary gland epithelial cells of all patients. Fluorescent quantitative polymerase chain reaction (FQ-PCR) and western blotting methods were adopted to detect and compare the transcriptional level of pIgR. The salivary gland epithelial cells of the 60 patients with oral bacterial infection were isolated and extracted, and they were divided into two groups (observation group and control group) randomly. Agonists were added to the observation group for acting for 24 h. FQ-PCR and immunofluorescence (IF) were adopted to detect and compare the transcriptional level of pIgR after acting with agonists. The toxicity of agonists on the cells was detected with Cell Counting kit-8 (CCK-8). The isolated salivary gland epithelial cells conformed to the morphology of epithelial cells, and adhered to the wall for growing. The transcriptional level of pIgR in the bacterial infection group was lower than that in the non-bacterial infection group (p<0.05). The transcriptional level of pIgR in the observation group was higher than that in the control group (p<0.05) after acting with agonists. Agonists can promote the rise of transcriptional level of pIgR in salivary gland epithelial cells, and the increase in pIgR is closely related to the cure of oral bacterial infection. Therefore, agonists can improve the oral immune function by regulating the transcription of pIgR. [ABSTRACT FROM AUTHOR]

Introduction: Streptococcus pneumoniae (S. pneumoniae) causes pneumococcal disease, which has high mortality and morbidity in children under two years of age, the elderly and immunocompromised individuals. This disease can be prevented by immunization, but the current vaccine, pili protein vaccine (PPV), is less likely to protect children under the age of two and only protects against the serotypes contained in the vaccine. Hence, a new vaccine is needed to enable full protection. The use of bacterial pili proteins may offer an alternative new vaccine. Therefore, the determination of the ability of such proteins to stimulate mucosal immunity with indicator expression of pIgR and s-IgA is required. Materials and Methods: Pili were isolated using the pili bacterial cutter method, and used for nasal vaccination to the rats. TGF-β1, IL-17A, and s-IgA were measured by ELISA while pIgR was examined by immunohistochemistry. Results: This study demonstrated that intranasal immunization with antigen (54 kDa pili protein) and antigen plus adjuvant significantly increased (p<0.05) the expression of TGF-β1. However, the expression of IL-17A increased significantly (p<0.05) only in rats immunized with antigen plus adjuvant. Further analysis demonstrated that intranasal immunization with antigen and antigen plus adjuvant significantly increased (p<0.05) expression of pIgR. Expression of sIgA in nasal lavage significantly increased (p<0.05) in those rats which had been immunized with pili protein plus adjuvant. Conclusion: This study showed that the immünization with S. pneumoniae pili protein increased expression of pIgR and sIgA which are important mucosal immunity components. Therefore, this protein has potency to be developed as nasal vaccination to prevent S. pneumoniae infection. [ABSTRACT FROM AUTHOR]

Background: Pneumococci are the major cause of bacterial meningitis globally. To cause meningitis pneumococci interact with the 2 endothelial receptors, polymeric immunoglobulin receptor (pIgR) and platelet endothelial cell adhesion molecule (PECAM-1), to penetrate the blood-brain barrier (BBB) and invade the brain.Methods: C57BL/6 mice were infected intravenously with bioluminescent pneumococci, and treated with ceftriaxone (1 hour postinfection) and anti-pIgR and PECAM-1 antibodies (1 or 5 hours postinfection), then monitored for 5 and 10 days. Bacterial brain invasion was analyzed using IVIS imaging and bacterial counts.Results: Ceftriaxone, given early after pneumococcal challenge, cleared pneumococci from the blood but not from the brain. After combining ceftriaxone with receptor blockade, using anti-pIgR and PECAM-1 antibodies, we found 100% survival after 5 and 10 days of infection, in contrast to 60% for ceftriaxone alone. Combined antibiotic and antibody treatment resulted in no or few viable bacteria in the brain and no microglia activation. Antibodies remained bound to the receptors during the study period. Receptor blockade did not interfere with antibiotic permeability through the BBB.Conclusions: We suggest that adjunct treatment with pIgR and PECAM-1 antibodies to antibiotics may prevent pneumococcal meningitis development and associated brain damages. However, further evaluations are required. [ABSTRACT FROM AUTHOR]

Dimeric IgA (dIgA) can move through cells via the IgA/IgM polymeric immunoglobulin receptor (PIGR), which is expressed mainly on mucosal epithelia. Here, we studied the ability of dIgA to target commonly mutated cytoplasmic oncodrivers. Mutation-specific dIgA, but not IgG, neutralized KRASG12D within ovarian carcinoma cells and expelled this oncodriver from tumor cells. dIgA binding changed endosomal trafficking of KRASG12D from accumulation in recycling endosomes to aggregation in the early/late endosomes through which dIgA transcytoses. dIgA targeting of KRASG12D abrogated tumor cell proliferation in cell culture assays. In vivo , KRASG12D-specific dIgA1 limited the growth of KRASG12D-mutated ovarian and lung carcinomas in a manner dependent on CD8+ T cells. dIgA specific for IDH1R132H reduced colon cancer growth, demonstrating effective targeting of a cytoplasmic oncodriver not associated with surface receptors. dIgA targeting of KRASG12D restricted tumor growth more effectively than small-molecule KRASG12D inhibitors, supporting the potential of this approach for the treatment of human cancers. [Display omitted] • Dimeric IgA undergoes bona fide transcytosis through PIGR+ carcinoma cells • Dimeric IgA specifically neutralizes mutated oncogenes inside tumor cells • Oncodrivers are expelled outside tumor cells through altered endosomal trafficking • Mutation-specific dimeric IgA specifically abrogates tumor growth in vivo Despite their long half-life, therapeutic antibodies are considered ineffective against intracellular antigens. Biswas et al. demonstrate that dimeric IgA undergoes transcytosis through PIGR+ epithelial cancer cells and can be engineered for the specific targeting of intracellular mutated oncodrivers. Binding of mutation-specific IgA expels oncodrivers from the cytoplasm and promotes anti-tumor immunity. [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]

Polymeric immunoglobulin receptor (pIgR) can bind and transport immunoglobulins (Igs), thus playing a role in mucosal immunity. In this study, pIgR gene was cloned in mandarin fish, Siniperca chuatsi , with the open reading frame (ORF) of 1011 bp, encoding 336 amino acids. The pIgR protein consists of a signal peptide, an extracellular domain, a transmembrane domain and an intracellular region, with the presence of two Ig-like domains (ILDs) in the extracellular domain, as reported in other species of fish. The pIgR gene was expressed in all organs/tissues of healthy mandarin fish, with higher level observed in liver and spleen. Following the immersion infection of Flavobacterium columnare , pIgR transcripts were detected in immune related, especially mucosal tissues, with significantly increased transcription during the first two days of infection. Through transfection of plasmids expressing pIgR, IgT and IgM, pIgR was found to be interacted with IgT and IgM as revealed by co-immunoprecipitation and immunofluorescence. • The pIgR gene was cloned in mandarin fish Siniperca chuatsi. • pIgR was expressed constitutively with higher level in liver and spleen. • High expression level of pIgR was observed in mucosal tissues following bacterial immersion. • Co-IP analysis showed that pIgR was reactive with IgT and IgM. [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:Secretory IgA (SIgA) and the polymeric immunoglobulin receptor (pIgR) have a pivotal role in gut homeostasis. Bovine lactoferrin (bLf) has been shown to modulate intestinal immunity and endogenous corticosterone. Considering the regionalization of the intestinal immune response, the aim of this work was to compare the impact of bLf on the IgA response in the proximal versus distal small intestine under physiological conditions. Methods:Groups of healthy male BALB/c mice were orally treated with one daily dose of bLf (50, 500, or 5000 μg) or untreated (control) for 7 d, and then sacrificed. From plasma samples, corticosterone levels were determined by an enzyme-linked immunosorbent assay (ELISA) kit. From distal and proximal segments of the small intestine, the following material was obtained: intestinal secretions to evaluate IgA levels by ELISA; epithelial cell extracts for protein-analysis of α-chain and pIgR by Western blot; mucosa samples for mRNA analysis of α-/J-chain, pIgR, and interleukin (IL)-2, -4, -5, and -6 by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR). Results:With 5000 μg of bLf, there were greater modulatory effects in the distal (versus proximal) segment, evidenced by an increase in the (i) level of total and specific IgA, (ii) protein expression of α-chain and pIgR, (iii) mRNA transcripts of α-chain, IL-2 and IL-5, and (iv) level of plasma corticosterone. Conclusions:Endogenous corticosterone elicited by bLf may have allowed for an IL profile that favored the IgA antibody response. The latter has a key role in maintaining intestinal homeostasis. [ABSTRACT FROM PUBLISHER]

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]

Problem The pIgR mediates transport of IgA into the lumen of mucosal tissues preventing pathogenic infection. Despite this, the expression of pIgR during chlamydial infections of the male and female reproductive tracts remains poorly understood. Method of study The expression of pIgR in response to hormone cycling or over the course of chlamydial infection was determined in vitro and in vivo by Western blot or immunohistochemistry. Results PIgR was upregulated in response to Chlamydia spp. infection of human epithelia, in both male and female mouse reproductive tracts. PIgR expression was found to be highest during estrus in the cervicovaginal and uterine epithelia and lowest during diestrus or following hormonal synchronization with Depo-Provera. Chlamydial infection of mice mediates upregulation of pIgR and transcytosis of IgA into the lumen. Conclusions Our results suggest that chlamydial infection enhances IgA secretion and pIgR expression by epithelia in the lower reproductive tracts of females and males, and hormone synchronization downregulates pIgR expression and transcytosis of IgA prior to challenge. [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]

A characteristic mucosal immune response involves the production of antigen-specific secretory immunoglobulin A ( SIgA) antibodies. In order to study transcytosis by mimicking the SIgA secretion and to screen for SIgA secretion-promoting substances, we developed a model system of a transfected Madin-Darby canine kidney (MDCK) cell line that expresses the human polymeric immunoglobulin receptor ( pIgR). We thus isolated the human dIgA (dimeric IgA)/pIgA (polymeric IgA) complex as the binding ligands. In the present study, a recombinant vector encoding the human pIgR gene was constructed and infected into MDCK cells. Following reverse transcription polymerase chain reaction (RT-PCR), immunoblotting, and immunofluorescence staining, we confirmed that pIgR was expressed in the transfectant MDCK-pIgR cells and was located at the basolateral side of the cell surface. We also confirmed the coexistence of the dIgA/pIgA complex in the IgA myeloma serum. The covalent dIgA/pIgA complex was then isolated from the serum of an IgA multiple myeloma patient using an ÄKTA purifier operation system with a HiPrep 16/60 Sephacryl S-300 HR column, in order to utilize the complex as transcytosis ligands for human pIgR. Finally, we confirmed the uptake of the isolated human dIgA/pIgA complex into MDCK-pIgR cells. We demonstrated that the human dIgA/pIgA complex was transcytosed into the apical side of the monolayer cells. Therefore, our MDCK-human pIgR cell transcytosis model is an operational system and can be used for screening functional food components that promote dIgA/pIgR transcytosis as well as SIgA secretion. [ABSTRACT FROM AUTHOR]

Stress is a response of the central nervous system to environmental stimuli perceived as a threat to homeostasis. The stress response triggers the generation of neurotransmitters and hormones from the hypothalamus pituitary adrenal axis, sympathetic axis and brain gut axis, and in this way modulates the intestinal immune system. The effects of psychological stress on intestinal immunity have been investigated mostly with the restraint/immobilization rodent model, resulting in an up or down modulation of SIgA levels depending on the intensity and time of exposure to stress. SIgA is a protein complex formed by dimeric (dIgA) or polymeric IgA (pIgA) and the secretory component (SC), a peptide derived from the polymeric immunoglobulin receptor (pIgR). The latter receptor is a transmembrane protein expressed on the basolateral side of gut epithelial cells, where it uptakes dIgA or pIgA released by plasma cells in the lamina propria. As a result, the IgA-pIgR complex is formed and transported by vesicles to the apical side of epithelial cells. pIgR is then cleaved to release SIgA into the luminal secretions of gut. Down modulation of SIgA associated with stress can have negative repercussions on intestinal function and integrity. This can take the form of increased adhesion of pathogenic agents to the intestinal epithelium and/or an altered balance of inflammation leading to greater intestinal permeability. Most studies on the molecular and biochemical mechanisms involved in the stress response have focused on systemic immunity. The present review analyzes the impact of stress (mostly by restraint/immobilization, but also with mention of other models) on the generation of SIgA, pIgR and other humoral and cellular components involved in the intestinal immune response. Insights into these mechanisms could lead to better therapies for protecting against pathogenic agents and avoiding epithelial tissue damage by modulating intestinal inflammation.