IgA Expression Research Articles

16945 IgA expression in adult cutaneous leukocytoclastic vasculitis and its effect on outcome

Background: Leukocytoclastic vasculitis (LCV) is a small vessel inflammatory disorder thought to be mediated by antibody-complex deposition. Previous studies have shown that patients with LCV and IgA deposition on direct immunofluorescence (DIF) are more likely to develop systemic co-morbidities and refractory disease, while other studies suggest that DIF in LCV has limited diagnostic value. Although certain cases may warrant DIF, data assessing IgA status in LCV patients and its impact on clinical outcome is lacking.

Lactococcus lactis NZ9000 Prevents Asthmatic Airway Inflammation and Remodelling in Rats through the Improvement of Intestinal Barrier Function and Systemic TGF-β Production

Introduction: The use of probiotics has been broadly popularized due to positive effects in the attenuation of aberrant immune responses such as asthma. Allergic asthma is a chronic respiratory disease characterized by airway inflammation and remodelling. Objective: This study was aimed to evaluate the effect of oral administration of Lactococcus lactis NZ9000 on asthmatic airway inflammation and lung tissue remodelling in rats and its relation to the maintenance of an adequate intestinal barrier. Methods: Wistar rats were ovalbumin (OVA) sensitized and challenged and orally treated with L. lactis. Lung inflammatory infiltrates and cytokines were measured, and remodelling was evaluated. Serum OVA-specific immunoglobulin (Ig) E levels were assessed. We also evaluated changes on intestinal environment and on systemic immune response. Results: L. lactis diminished the infiltration of proinflammatory leucocytes, mainly eosinophils, in the bronchoalveolar compartment, decreased lung IL-4 and IL-5 expression, and reduced the level of serum allergen-specific IgE. Furthermore, L. lactis prevented eosinophil influx, collagen deposition, and goblet cell hyperplasia in lung tissue. In the intestine, L. lactis-treated asthmatic rats increased Peyer’s patch and goblet cell quantity and mRNA expression of IgA, MUC-2, and claudin. Additionally, intestinal morphological alterations were normalized by L. lactis administration. Splenocyte proliferative response to OVA was abolished, and serum levels of transforming growth factor (TGF)-β were increased by L. lactis treatment. Conclusions: These findings suggest that L. lactis is a potential candidate for asthma prevention, and the effect is mediated by the improvement of intestinal barrier function and systemic TGF-β production.

Prolonged exposure to high humidity and high temperature environment can aggravate influenza virus infection through intestinal flora and Nod/RIP2/NF-κB signaling pathway

Seasonal influenza is an acute viral infection caused by influenza virus, which is often prevalent in the summer and winter. The influenza virus can infect pigs and poultry. Some literature reports that the influenza virus has an outbreak in summer. The summer climate is characterized by a high humidity and high temperature environment, which is the same as many animal feeding and growing environments. We established a flu animal model under a high temperature and humidity environment during the day to observe the impact of high humidity and high temperature environment on the mice after contracting the influenza virus. Our results indicate that the intestinal flora of 16 s rDNA cultured in High humidity and high temperature environment changes, the intestinal mucosal permeability increases, the expression of pIgR, sIgA, and IgA in the intestinal mucosal immune system decreases, and the NLR immune recognition signaling pathway NOD1 is activated. The expression of related genes such as NOD2, NF-κB, and pIgR increases, which leads to the increase of related inflammatory factors in the vicinity of the intestines, aggravating local inflammation. High humidity and high temperature environment can cause the expression of inflammatory cytokines in the body to rise, causing Th17/Treg immune imbalance, inhibiting Treg maturation and differentiation, and increasing the expression of IL-2, IL-6, and other cytokines, while the expression of IFN-γ and IL-17A decreases. This condition worsens after infection with the influenza virus. Overall, our study found that High humidity and high temperature environment affect the intestinal flora and the body's immune status, thereby aggravating the status of influenza virus infection.

Influence of Lacto-Immuno-Vital on growth performance and gene expression of IgA, MUC-2, and growth factor IGF-2 in the jejunum of broiler chickens

The effects of Lacto-Immuno-Vital synbiotic preparation on gene expression of IgA, MUC-2, and growth factor IGF-2 in the jejunum and on BW gain in broiler chickens were studied. A flock of 64,400 1-day-old Hybrid ROSS 308 chickens was inducted in the 42-day experiment. The chickens were divided into 2 equally size groups in separate halls. The chickens in the experimental (E) group received 500 g of Lacto-Immuno-Vital in 1,000 L of drinking water. The preparation was administered daily from the first day (day 1) to day 7 of the experiment. From day 7 to day 22, it was given in pulsed manner (every third day) at a dose of 300 g in 1,000 L of drinking water. The broiler chickens in the E group gained more weight (P < 0.001) compared with control from day 10 to day 42. Death of animals during feeding period was 1,078 chickens in the E group compared with 1,115 dead chickens in the control group. Feed conversion ratio was 1.61 kg of supplemented diet/kg of BW in the E group compare with 1.67 kg of nonsupplemented diet/kg of BW in control. The relative expression of IgA gene in the jejunum was upregulated on day 22 in the E group compared with control (P < 0.05), whereas relative expression of MUC-2 gene was upregulated in the E group compared with control on day 8 and day 22 (P < 0.05; P < 0.001). Similarly, relative expression of IGF-2 gene was upregulated in the E group compared with control on both samplings (P < 0.01). The composition of Lacto-Immuno-Vital synbiotic preparation showed beneficial effects on growth performance, feed conversion ratio, morbidity, mortality, and selected parameters of mucosal immunity in the chicken jejunum.

The Potential Role of Immune Alteration in the Cancer-COVID19 Equation-A Prospective Longitudinal Study.

Background: The risk of cancer patients to develop COVID19 infection is unclear. We aimed to prospectively study cancer patients and oncology healthcare workers for COVID19 serology. In IgG+ cases, immune profile was determined to portray the pattern of immune response to SARS-CoV2. Methods: Cancer patients on active treatment and healthcare workers were enrolled. During the study period (3/2020-6/2020), demographic data and blood were collected at three time points. Expression of IgG, IgM, and IgA were assessed. In SARS-CoV-2 IgG+ cases and matched negative cases, we performed mass cytometry time of flight (CyTOF) analysis on the basis of the expression of surface markers. Results: The study included 164 cancer patients on active intravenous treatment and 107 healthcare workers at the cancer center. No symptomatic cases were reported during the study period. Serology analysis revealed four IgG+ patients (2.4%) and two IgG+ healthcare workers (1.9%)-all were asymptomatic. CyTOF analysis demonstrated substantial reduction in myeloid cells in healthcare workers who were SARS-CoV-2 IgG+ compared to those who were SARS-CoV-2 IgG-, whereas in cancer patients, the reduction was relatively milder (≈50% reduction in SARS-CoV-2 IgG+ cancer patients compared with ≈90% reduction in SARS-CoV-2 IgG+ workers). Conclusion: Our results indicate a similar rate of asymptomatic COVID19 infection in cancer patients and healthcare workers in a longitudinal study throughout the pandemic time. Due to differential immune cell profiles of cancer patients who are treated with immunomodulatory agents, the host response to the SARS-COV2 may play a role in COVID19 course and representation. The immunological perspective of cancer treatments on the risk for COVID19 infection should be further explored.

Bioinformatics Analysis of Gut Microbiota and CNS Transcriptome in Virus-Induced Acute Myelitis and Chronic Inflammatory Demyelination; Potential Association of Distinct Bacteria With CNS IgA Upregulation.

Virus infections have been associated with acute and chronic inflammatory central nervous system (CNS) diseases, e.g., acute flaccid myelitis (AFM) and multiple sclerosis (MS), where animal models support the pathogenic roles of viruses. In the spinal cord, Theiler's murine encephalomyelitis virus (TMEV) induces an AFM-like disease with gray matter inflammation during the acute phase, 1 week post infection (p.i.), and an MS-like disease with white matter inflammation during the chronic phase, 1 month p.i. Although gut microbiota has been proposed to affect immune responses contributing to pathological conditions in remote organs, including the brain pathophysiology, its precise role in neuroinflammatory diseases is unclear. We infected SJL/J mice with TMEV; harvested feces and spinal cords on days 4 (before onset), 7 (acute phase), and 35 (chronic phase) p.i.; and examined fecal microbiota by 16S rRNA sequencing and CNS transcriptome by RNA sequencing. Although TMEV infection neither decreased microbial diversity nor changed overall microbiome patterns, it increased abundance of individual bacterial genera Marvinbryantia on days 7 and 35 p.i. and Coprococcus on day 35 p.i., whose pattern-matching with CNS transcriptome showed strong correlations: Marvinbryantia with eight T-cell receptor (TCR) genes on day 7 and with seven immunoglobulin (Ig) genes on day 35 p.i.; and Coprococcus with gene expressions of not only TCRs and IgG/IgA, but also major histocompatibility complex (MHC) and complements. The high gene expression of IgA, a component of mucosal immunity, in the CNS was unexpected. However, we observed substantial IgA positive cells and deposition in the CNS, as well as a strong correlation between CNS IgA gene expression and serum anti-TMEV IgA titers. Here, changes in a small number of distinct gut bacteria, but not overall gut microbiota, could affect acute and chronic immune responses, causing AFM- and MS-like lesions in the CNS. Alternatively, activated immune responses would alter the composition of gut microbiota.

IL-21 and IL-5 coordinately induce surface IgA+ cells

Intestinal IgA is induced by microbes and food antigens. Peyer’s patches (PPs) are known as one of the inductive sites for intestinal IgA production. However, the precise mechanism of IgA induction is as yet unknown. IgA secretion was induced from IgD+ B cells in vitro by stimulus with lipopolysaccharide in the presence of only retinoic acid (RA) and low doses of TGF-β1. Surface IgA+ cells were effectively induced from IgD+ B cells in vitro by the mixture of RA and the cytokines TGF-β1, APRIL, IL-5 and IL-21. rIL-21 upregulated surface IgA+ but impaired the proliferation of stimulated B cells in the presence of rTGF-β1, RA and rAPRIL, in vitro. The addition of rIL-5 restored the impaired proliferation by rIL-21, resulting in the expansion of IgA+ cells. rIL-21 induced the expression of Aicda and Prdm1, and impaired Rel in IgD+ B cells. Blockade of IL-21R signaling by a neutralizing mAb in vivo led to lower frequencies of IgA+ and IgG2b+ cells and lower germinal center B cells in PPs in a homeostatic condition. Although amounts of small intestinal IgA and titers of anti-dsDNA, the major target of intestinal IgA, in these mice were not altered, anti-OVA IgA titers induced by OVA drinking in OVA-specific T-cell receptor (TCR) transgenic mice were decreased. PP-deficient TCR transgenic mice showed diminished anti-OVA IgA induction. Blockade of IL-5R signaling in vivo led to similar results with relatively weaker effects than that of IL-21R mAb administration. These results suggest that IL-21 and IL-5 play cooperative roles in surface expression of IgA in PPs.

Complete ablation of tumor necrosis factor decreases the production of IgA, IgG, and IgM in experimental central nervous system tuberculosis.

Objective(s):This study aimed to explore the contribution of tumor necrosis factor (TNF) in the recruitment of B-cell and secretion of immunoglobulins (Igs) during cerebral tuberculosis (TB).Materials and Methods:In this work, the contributing role of TNF in regulating Ig secretions was investigated by comparing wild type TNF (TNFf/f), B-cell-derived TNF (BTNF-/-), and complete TNF ablation (TNF-/-) in a mouse cerebral Mycobacterium tuberculosis infection. Using flow cytometry and ELISA, we were able to examine the recruitment of B-cell subsets, and the production of Igs; also assessed the expression of surface markers on B cell subsets. Results:Here, we found that TNF-/- mice showed defective expression of IgA, IgG, and IgM antibodies compared with TNFf/f and BTNF-/- mice, which was significantly decreased in the expression of surface markers and co-stimulatory molecules. Moreover, mice that produced low antibody levels were not able to control infection, therefore progressed to disease; providing direct evidence for the TNF gene-regulating humoral immunity during central nervous system (CNS) M. tuberculosis infection. In contrast, BTNF-/- mice controlled the infection and had levels of IgA, IgG, and IgM comparable to TNFf/f mice.Conclusion:Together, our results demonstrate that TNF may serve as an essential regulator of antibody-mediated immune responses in CNS TB. However, the protective level exhibited by TNF-producing B cells could be defined as baseline protection that could be used in the development of new therapeutic targets or designing new vaccines.

Human IgA Monoclonal Antibodies That Neutralize Poliovirus, Produced by Hybridomas and Recombinant Expression.

Poliovirus (PV)-specific intestinal IgAs are important for cessation of PV shedding in the gastrointestinal tract following an acute infection with wild type or vaccine-derived PV strains. We sought to produce IgA monoclonal antibodies (mAbs) with PV neutralizing activity. We first performed de novo IgA discovery from primary human B cells using a hybridoma method that allows assessment of mAb binding and expression on the hybridoma surface: On-Cell mAb Screening (OCMS™). Six IgA1 mAbs were cloned by this method; three potently neutralized type 3 Sabin and wt PV strains. The hybridoma mAbs were heterogeneous, expressed in monomeric, dimeric, and aberrant forms. We also used recombinant methods to convert two high-potency anti-PV IgG mAbs into dimeric IgA1 and IgA2 mAbs. Isotype switching did not substantially change their neutralization activities. To purify the recombinant mAbs, Protein L binding was used, and one of the mAbs required a single amino acid substitution in its κ LC in order to enable protein L binding. Lastly, we used OCMS to assess IgA expression on the surface of hybridomas and transiently transfected, adherent cells. These studies have generated potent anti-PV IgA mAbs, for use in animal models, as well as additional tools for the discovery and production of human IgA mAbs.

Cytokine Gene Expression and IgA Production in the Spleen of Chickens Infected with Eimeria tenella

To explore the effect of Eimeria tenella infection on the cytokines gene expression and IgA production in the spleen of chickens, the morphological characteristics of the spleen were observed through optical and transmission electron microscopy. The IgA production was determined through immunohistochemistry. The mRNA expression levels of splenic cytokines were detected through real-time PCR. Compared to the control group, along with the infection of E. tenella, the splenic lymphocytes exhibited irregular and cracked membranes, mitochondria swelled even vacuolization, the IgA expression in spleen tissue was decreased by 55.57% (p lessthan 0.01). Likewise, the mRNA expression levels of IL-2 and IL-1â decreased by 40% (plessthan 0.01) and 43% p lessthan 0.05), respectively. By contrast, the IL-6, IFN-g and IL-10 levels increased by 158% (p lessthan 0.01), 464% (p lessthan 0.05) and 379% p lessthan 0.01), respectively. These results indicated that the spleen implement an important function in the antagonism of E. tenella, which suggest a new strategy to control coccidiosis by improving the peripheral immunity of chickens.

Immunoglobulin receptors expression in indian colon cancer patients and healthy subjects using a noninvasive approach and flowcytometry

Background:Isolation of viable colonocytes from human stool is a noninvasive and convenient approach that can be used for diagnostic, screening, management, and research on various gastrointestinal (GI) diseases including colon cancer. Limited studies are available globally and for the first time in this article, we have reported the immunoglobulin (Ig) (IgA and IgG) receptors concentration on viable colonocytes for Indian colon cancer patients using this noninvasive approach.Materials and Methods:Viable colonocytes from stool were isolated by the Somatic Cell Sampling and Recovery method (Noninvasive Technology, USA) and processed for the assessment of Igs (IgA and IgG) receptors expression using standard immunophenotyping and flow cytometry.Results:IgA and IgG receptor expression was measured and reported on these viable colonocytes. There was a significant difference in the expression of IgA and IgG receptors on viable colonocytes between colon cancer patients and healthy individuals.Conclusion:This noninvasive technique is a promising approach for the detection of molecular and immunological markers that will help clinicians in the diagnosis, screening, monitoring, and management of different GI diseases including colon cancer.

Altered microbial community structure in PI3Kγ knockout mice with colitis impeding relief of inflammation: Establishment of new indices for intestinal microbial disorder

Lipopolysaccharide stimulates the intestinal microbiome to activate phosphoinositide 3 kinase (PI3K) signaling via several pathways; however, the direct effect that PI3K has on the intestinal bacterial community remains unclear. Herein, we investigate changes in the colonic microbiome of colitis PI3Kγ-knockout (PI3Kγ−/−) mice. Additionally, the effect of anal administration of colonic irrigation fluid from control mice to those with colitis was examined. Microbial 16S rRNA genes from the colonic mucosa of PI3Kγ−/− and WT mice were sequenced using Illumina MiSeq platform, and colonic IgA, IL-2, IL-10, and IL-17A production was quantified by western blot analysis. Myeloperoxidase (MPO) activity was detected by absorbance via colorimetric analysis. From the results, two new indices were derived by dividing the bacterial community into invading taxa, common taxa, and vanishing taxa. These indices were used to estimate the degree of microbiome disorder in chronic experimental colitis models. PI3Kγ−/− mice showed slower remission of inflammation as assessed by the disease activity index，pathological score, IL-2, IL-17, IL-10, IgA expression and MPO activity. The unique and common taxa of wild-type and PI3Kγ−/− mice increased as colitis symptoms regressed. Continuous loss of commensal bacteria happened with the continuous invasion of exogenous bacteria in the intestinal mucosa of PI3Kγ‐−/− mice after colitis begin to aggravate. However, transplantation of normal intestinal microbiota to PI3Kγ−/− mice promoted remission of inflammation; while the microbial dysbiosis observed during PI3Kγ dysfunction aggravated the intestinal microbiome disorder and impeded colitis recovery. Thus, the PI3Kγ signaling pathway may regulate microbial community composition in the colon.

The Influence of Ketogenic Diets on Psoriasiform-Like Skin Inflammation

The Influence of Ketogenic Diets on Psoriasiform-Like Skin Inflammation

Clinical and morphological features of allergic enterocolitis in young children

The purpose was to study the clinical and morphological features of allergic enterocolitis in young children.Material and methods. 29 patients aged from 2 months to 2 years with severe allergic enterocolitis, who underwent endoscopic examination with targeted biopsy of the intestinal mucosa, were treated. For comparison of morphological, histochemical and immunohistochemical features, intestinal mucosal biopsies of children of the same age with gastrointestinal disorders, caused by protracted or chronic course of non-specific non-ulcer process in the intestine, were used. Immunohistochemical studies were performed on serial paraffin sections in accordance with standard protocols using Ab68 (Macrophage Marker) Ab-3 (Clone KP1), IgA (Heavy Chain) Ab-2, IgE (Epsilon-Heavy Chain) Ab-1 monoclonal antibodies and Imaging Ultra Vision Quanto Detection System HRP DAB (USA).Results. The debut of gastrointestinal symptoms in young children with allergic enterocolitis was observed in the first months of life and was characterized by vomiting or persistent regurgitations, intense griping, diarrhea with large amounts of mucus and/or blood, and delayed physical development. Presence of inflammatory process of intestinal mucosa with eosinophilic infiltration was morphologically established. According to immunohistochemical study, expression of CD68-macrophages and increased expression of IgA and IgE in the duodenal mucosa were detected.Conclusions. The diagnosis of allergic enterocolitis at an early age is complicated by the absence of specific clinical gastrointestinal symptoms and non-informative allergic examination (IgE-independent mechanism of inflammation), which may require morphological study to verify the diagnosis. Morphologically, in the examined patients, not only the presence of inflammatory process of the mucous membrane of the intestine with expressed eosinophilic infiltration is established, but also the immunohistochemistry data revealed the increased expression of CD68-macrophages, IgA and IgE in the mucous membrane of the duodenum, indicating the activation of both cellular and local humoral immunity.

Radiation therapy-induced reactive oxygen species specifically eliminates CD19+IgA+ B cells in nasopharyngeal carcinoma.

PurposeNasopharyngeal carcinoma (NPC) is one of the most common head and neck cancers and is thought to be related to the mucosal immune system. Radiation therapy (RT) is the primary treatment for NPC due to the high radiosensitivity of cancer cells. However, little is known about the impact of RT on the mucosal immune system.Patients and methodsIn this study, the expression of immune markers CD19, CD24, CD27, CD8, and IgA before and after RT, were analyzed using flow cytometry. Cytokines were assessed using the enzyme-linked immunosorbent assay. Reactive oxygen species (ROS) was assayed by flow cytometry and fluorescence staining using 2ʹ,7ʹ -dichlorofluorescein diacetate.ResultsWe found that primary NPC patients had a significant increase in CD19+CD138−IgA+ B cells, which was then decreased after RT. Interestingly, the changes in CD19+CD138−IgA+ B cell frequency was accompanied by corresponding frequency changes in cytotoxic T cells (CTL), which are powerful anti-tumor lymphocytes. Mechanistically, we found that ROS release during RT specifically eliminated CD19+CD138−IgA+ B cells.ConclusionThese findings suggest that RT may regulate the immune system and opens up new avenues for the utilization of immune-radiotherapy in NPC.

PS1266 CLINICAL ASPECTS, MORPHOLOGICAL ANALYSIS AND IMMUNOHISTOCHEMICAL CHARACTERISTICS OF PATIENTS WITH FOLLICULAR LYMPHOMA IN DUODENUM BIOPSY: CASES WITH RESTRICT DISEASE VERSUS ADVANCED PRESENTATION

Background:Duodenal‐type follicular lymphoma (DFL) is recognized as a distinctive entity by the 2016 WHO classification. The largest series to date described 63 cases, highlighting its indolent form and good prognosis. Despite being defined as restricted to the duodenum, progressions can occur. The differentiation from typical follicular lymphoma with duodenum involvement is still a matter for debate.Aims:To study the pathological and clinical aspects of patients with follicular lymphoma (FL) with duodenal involvement, comparing cases restricted to duodenum with cases presenting with advanced disease.Methods:Retrospective analysis of patients diagnosed with FL by endoscopic duodenal biopsies. Cases reviewed for biopsied layers, architectural pattern, follicular dendritic cell pattern (FDCP), Ki‐67 and IgA expression by neoplastic cells. Patients were divided into restrict duodenum involvement (RDI) and with extra duodenal involvement (EDI), by clinical staging at diagnosis.Results:A total of 26 patients, 50% male and average age of 58 years (39–86). Twenty‐one had pathological revision but 2 did not complete clinical staging and were removed from analysis. In the 19 reviewed, 5 (26%) were with EDI and 14 (74%) with RDI. None patient with EDI had mucosa‐restricted disease, but that feature occurred in 9 (64%) patients with RDI. RDI group presented follicular architecture in 9 patients (64%) and 5 (36%) was follicular and diffuse pattern, while 1 (20%) of the EDI group had follicular architecture and 4 (80%) follicular and diffuse. The so‐called “duodenal pattern”, related to FDCP, was reported in 12 (85%) patients with RDI against 3 (60%) with EDI. KI‐67 varied between 5 – 50% in RDI (average 18%) and between 15 – 40% in EDI (average 22%). There was IgA expression in 9 (64%) patients with RDI and in only 1 (20%) patient with EDI. The figure below illustrates IgA expression. Concerning the clinical aspects, from the 26 patients, 19 (73%) had features in the second portion of the duodenum and 16 (61%) the endoscopic lesions were small white nodules. Of the 18 patients that also had a colonoscopy, 1 had lesions in the distal ileum and other in the colon. At diagnosis, endoscopy was made in 9 (35%) patients as screening and in 12 (46%) over gastrointestinal symptoms (GIS). Most patients were FLIPI 0 or 1 (58%) and all had low grade 1–2 FL. For the treatment analysis, 4 patients were excluded because of insufficient data. In the 22 remained patients, the majority was treated with rituximab (R) monotherapy (n = 10, 45%), followed by watch and wait (36%), chemotherapy (14%) and radiotherapy (5%). Indication of treatment was clear in 14 patients, being GIS the principal one (50%). R maintenance was chosen in 8 (36%) patients, 6 previously treated with R and 2 with chemotherapy. All patients treated achieved complete remission by endoscopy and biopsy but one, assigned to R and with partial remission. There was 1 patient in watch and wait that had a spontaneous remission. With a median follow up of 14.5 months (1–40), none patient died and only 1 patient has progressed (EDI group and with nodal disease).Summary/Conclusion:The RDI group (related to the WHO definition of DFL) had more localized mucosal disease, predominantly with the follicular pattern and with higher expression of IgA. The “duodenal pattern” and Ki‐67 did not seem to separate between RDI and EDI patients. Rituximab monotherapy and watch and wait strategy showed good results in this population. A larger cohort of patients with longer follow up may assess better these results.image

Expressions of CD3 and CD4 T Cells in Peripheral Blood of Patients with Immunoglobulin A Nephropathy and Their Clinical Significance

Immunoglobulin A nephropathy (IgAN) is a primary glomerulonephritis characterized by abnormal immune response-mediated deposition of polymeric IgA (pIgA) in mesangium. As a type of important immune cells, the relationship of CD3 or CD4 with the pathogenesis of IgAN remains poorly understood. In this study, 38 patients with IgAN, 7 patients with idiopathic membranous nephropathy (MN) and 46 healthy adults without history of kidney disease were enrolled. Peripheral blood was collected for further evaluation of the expressions of CD3 and CD4 and IgA by flow cytometry, quantitative polymerase chain reaction (qPCR) and Western blot. Meanwhile, the expression of IgA was detected by ELISA. The result showed that expression of CD3 T cells was down-regulated in patients with IgAN, while amounts of CD4 T cells and IgA level were significantly increased compared to normal control (P &lt; 0.05). However, no signficant changes in CD3, CD4 T cells were found in patients with MN. Our study demonstrates that CD3 and CD4 T cells as well as IgA are involved in the pathogenesis of IgAN and these targets might be beneficial for the treatment of IgAN.

Activation of aryl hydrocarbon receptor with structurally diverse ligands represses class switch to immunoglobulin A

Abstract Activation of the aryl hydrocarbon receptor (AhR) with its prototypical agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) represses activation-induced cytidine deaminase expression and B cell class switch recombination (CSR). Structurally diverse AhR agonists have been shown to have differing impacts on T cell differentiation. We hypothesized that AhR activation with structurally diverse agonists would have differential impacts on CSR in B cells. The following agonists were tested: the endogenous ligands cinnabarinic acid (CA), 5,11-dihydroindolo[3,2-b]carbazole-6-carboxaldehyde (FICZ), and kynurenine (Kyn); the pharmaceuticals omeprazole (Omep) and tamoxifen (Tam); and the dietary component indole-3-carbanol (I3C). Mouse splenic CD19+ B cells were cultured with LPS, α-IgD-dextran, and TGF-β1 to stimulate CSR to IgA. IgA expression was measured by flow cytometry. Relative to control cells (10.84% IgA+), exposure to CA, FICZ, I3C, Kyn, Omep, Tam, or TCDD significantly decreased the proportion of IgA+ cells (P &amp;lt; 0.05) to 4.48%, 2.48%, 2.25%, 4.39%, 0.79%, 6.95% and 3.93%, respectively. In contrast, CH-223191, an AhR antagonist, significantly increased the proportion of IgA+ cells to 16.19%. These results suggest that repression of CSR is a common effect of AhR agonists in activated mouse B cells.

Zinc source modulates intestinal inflammation and intestinal integrity of broiler chickens challenged with coccidia and Clostridium perfringens

Two dietary sources of zinc (ZnSO4 or organic Zn) were tested in chickens challenged with coccidiosis (Co) or coccidiosis plus Clostridium perfringens (CoCPF). On day 14, the chickens were orally gavaged with ∼5,000 Eimeria maxima sporulated oocysts. On day 19, 20, and 21 chickens challenged with C. perfringens were given a broth culture containing 108 cfu of this bacterium. Productive performance parameters were determined at d 14, 21, and 28. On day 21, necrotic enteritis (NE) lesions were scored, and intestinal permeability was evaluated. Jejunum and cecal tonsils were collected for morphology and gene expression analysis. On day 21, organic Zn improved BW gain by 18.6% (P = 0.07), and FCR by 12% (P = 0.09) in CoCPF challenged chickens vs. birds fed ZnSO4. From 1 to 28, organic Zn increased BW gain (P = 0.02), and improved FCR (P = 0.03) vs. birds fed ZnSO4. At 21 d, NE lesions were only observed in CoCPF birds (P < 0.001), and mortality due to NE was only observed when CoCPF birds were fed ZnSO4 (P = 0.001). Organic Zn fed birds had increased villus height in the jejunum (P = 0.005) and decreased intestinal permeability (P = 0.01) vs. ZnSO4. In the jejunum, organic Zn fed birds showed a downregulation of expression of IL-8 (P = 0.02), and upregulation of IL-10 (P = 0.05) in CoCPF birds vs. ZnSO4- CoCPF birds. As main effect, birds supplemented with organic Zn had higher mRNA expression of TLR-2 (P = 0.02) and IgA (P = 0.01). In the cecal tonsils, organic Zn fed birds showed upregulation of iNOS (P = 0.008) in CoCPF birds vs. ZnSO4-CoCPF birds. Organic Zn supplementation reduced intestinal permeability and attenuated intestinal inflammation of broilers co-challenged with coccidia and C. perfringens.

Transformation strategies for stable expression of complex hetero-multimeric proteins like secretory immunoglobulin A in plants.

SummaryPlant expression systems have proven to be exceptional in producing high‐value complex polymeric proteins such as secretory IgAs (SIgAs). However, polymeric protein production requires the expression of multiple genes, which can be transformed as single or multiple T‐DNA units to generate stable transgenic plant lines. Here, we evaluated four strategies to stably transform multiple genes and to obtain high expression of all components. Using the in‐seed expression of a simplified secretory IgA (sSIgA) as a reference molecule, we conclude that it is better to spread the genes over two T‐DNAs than to contain them in a single T‐DNA, because of the presence of homologous recombination events and gene silencing. These T‐DNAs can be cotransformed to obtain transgenic plants in one transformation step. However, if time permits, more transformants with high production levels of the polymeric protein can be obtained either by sequential transformation or by in‐parallel transformation followed by crossing of transformants independently selected for excellent expression of the genes in each T‐DNA.