Number Of IgA-positive Cells Research Articles

Investigating mammary glands of lactating goats for the presence of tertiary lymphoid organs.

Ectopic tertiary lymphoid organs (TLOs) have been identified in many organs, such as the lungs, nasal cavities, and kidneys of both mice and humans. Although lymphocyte aggregates have been observed in the mammary glands of ruminants, the details remain unclear. In this study, we investigated the mammary glands of lactating goats for the presence of TLOs. The localization of CD20 (B cells), CD3 (T cells), MECA79 (high endothelial venules), CD40 (follicular dendritic cells), BCL6 (germinal center), and IgA was examined by immunohistochemistry. The concentrations of IgG, IgA, lactoferrin, β-defensin-1, cathelicidin-2, cathelicidin-7, S100A7, and S100A8 in milk were measured by ELISA. The localization and amount of tight junction (TJ) proteins (claudin-3 and claudin-4) were examined using immunofluorescence and western blotting. We found that 19 out of 30 udders contained lymphocyte aggregates, which showed positive reactions against CD20, CD3, CD40, and MECA79. In addition, large-sized aggregations showed separate localization of B cells and T cells and a positive reaction against BCL6, although BCL6 was sparsely localized in the aggregations. These results indicate that mammary glands of lactating goats contain TLOs. The IgG and IgA concentrations in the milk of TLO-positive goats and the number of IgA-positive cells were higher than those in negative goats. Furthermore, claudin-4 was localized in the TJ region and the amount was higher in TLO-positive mammary glands than that in the negative group, indicating the presence of leakages at TJs. In conclusion, a majority of lactating goat udders have TLOs, which contribute to local immunity by producing immunoglobulins.

Local Heat Treatment of Goat Udders Influences Innate Immune Functions in Mammary Glands.

Heat stress and mastitis adversely affect milk production in dairy ruminants. Although the udder temperature is elevated in both conditions, the influence of this local temperature rise on milk production and immune function of ruminant mammary glands remains unclear. To address this question, we heated the mammary glands of goats by covering one half of the udder with a disposable heating pad for 24h, the other uncovered half served as a control. Sixteen Tokara goats (1-5 parity) and three Shiba goats (1-2 parity) at the mid-lactation stage were individually housed, fed 0.6kg of hay cubes and 0.2kg of barley per day, and had free access to water and trace-mineralized salt blocks. Milk samples were collected every 6h for 24h after covering (n = 16), and deep mammary gland tissue areas were collected after 24h (n = 5). The concentrations of antimicrobial components [lactoferrin, β-defensin-1, cathelicidin-2, cathelicidin-7, and immunoglobulin A (IgA)] in milk were measured by the enzyme-linked immunosorbent assay (ELISA). The localization of IgA was examined by immunohistochemistry. The mRNA expression and protein concentrations of C-C motif chemokine ligand-28 (CCL-28) and interleukin (IL)-8 in the mammary gland tissue were measured using quantitative polymerase chain reaction and ELISA, respectively. The somatic cell count in milk was significantly higher in the local heat-treatment group than in the control group after 12h of treatment. The treatment group had significantly higher concentrations of cathelicidin-2 and IgA than the control group after 24h of treatment. In addition, the number of IgA-positive cells in the mammary stromal region and the concentration of CCL-28 in the mammary glands were increased by local heat treatment. In conclusion, a local rise in udder temperature enhanced the innate immune function in mammary glands by increasing antimicrobial components.

Immunomodulation and Intestinal Morpho-Functional Aspects of a Novel Gram-Negative Bacterium Rouxiella badensis subsp. acadiensis

A novel bacterium (Rouxiella badensis subsp. acadiensis) isolated from the microbiota of wild blueberry fruit was investigated for its immunomodulation capabilities and intestinal morpho-functional aspects. The whole-genome shotgun sequencing of this bacterium led to its new taxonomy and showed absence of pathogenicity genes. Although the bacterium was used for blueberry-fermentation and enhancing its anti-inflammatory effects on neurodegeneration, diabetes, and cancer, no study has assessed the effect of the bacterium on health. In this study, we used several in vitro and in vivo assays to evaluate the interaction of R. badensis subsp. acadiensis with the intestinal mucosa and its impact on the localized immune response. The strain antibiotic susceptibility has been investigated as well as its tolerance to gastric and intestinal environment and ability to attach to human intestinal epithelial cells (Caco-2 and HT-29). In addition, Balb/c mice were used to explore the immune-modulatory characteristics of the live bacterium at the intestinal level and its impact on the morpho-functional aspects of the intestinal mucosa. In vitro assays revealed the ability of R. badensis subsp. acadiensis to survive the gastric and intestinal simulated conditions and to satisfactorily adhere to the human intestinal epithelial cells. The bacterium was shown to be sensitive to an array of antibiotics. Immuno-modulation studies with mice orally administered with R. badensis subsp. acadiensis showed a higher number of IgA positive cells in the small intestine, a higher concentration of the anti-inflammatory cytokine IL-10 in the intestinal mucosa, as well as an increase in the number of goblet cells. The anti-inflammatory cytokine miR146a was found to be increased in the ileum and brain. Furthermore, it increases the number of goblet cells which contribute to intestinal barrier integrity. Taken together, our findings reflect the ability of the tested bacterium to modulates the intestinal homeostasis and immune response. Detailed safety unpublished studies and genome data support our finding. The strain Rouxiella badensis subsp. acadiensis has been filed in a provisional patent; a U.S. Provisional Application No. 62/916,921 entitled “Probiotics Composition and Methods.” Future studies are still needed to validate the potential utilization of this strain as functional food and its potential probiotic effect.

Effects of colostrum whey on immune function in the digestive tract of goats.

The objective of the present study is to examine whether colostrum whey can have an effect on immune function in goats digestive tract. Two milliliters of colostrum whey (colostrum group) or water (control group) were administrated orally to goats every day for 3weeks. Blood was collected twice a week for 3weeks to measure immunoglobulin A (IgA), interleukin 8 (IL-8), and IL-10. At the end of the experimental period, the parotid glands, oral mucosa, lingua, esophagus, jejunum, and ileum were collected for immunohistochemical detection of IgA, cathelicidin-7, and S100A8. The ratio of the length of IgA-positive mucosal surface in the esophagus to the total esophageal length was significantly greater in the colostrum group than in the control group. The number of IgA-positive cells in the labial gland and ileum in the colostrum group was significantly higher than that in the control group. There were no significant differences between the colostrum and control groups in the number of cathelicidin-7-positive cells in the jejunum and ileum and in the number of S100A8-positive cells in the lingua, jejunum, and ileum. These results suggest that colostrum stimulates the recruitment of plasma cells into the labial gland, which then secrete more IgA into the saliva.

Lack of Correlation between Mucosal Immunoglobulin A-positive Plasma Cell Numbers and TLR5 Genotypes in German Shepherd Dogs with Idiopathic Chronic Enteropathy

It has been suggested previously that a deficiency in mucosal immunoglobulin (Ig) A production could be involved in the pathogenesis of chronic enteropathy in German shepherd dogs (GSDs). Recent research has shown that single nucleotide polymorphisms in the gene encoding Toll-like receptor (TLR)-5 are associated with an increased risk of development of chronic idiopathic enteropathy in this breed. IgA is essential for mucosal immunity and studies in mice have linked the interaction of TLR5 with its ligand flagellin to class switching of B cells into IgA-producing plasma cells. We hypothesized that dogs carrying the risk-associated (RA) genotypes for G22A and C100T genes of TLR5 would have a different number of IgA plasma cells in the duodenal and colonic mucosa compared with dogs carrying the risk-protective (RP) genotypes. Thirty-one GSDs were diagnosed with idiopathic chronic enteropathy by clinical exclusion diagnosis and histopathological confirmation. Immunohistochemistry was performed using goat anti-dog IgA primary antibody. Two sections of duodenum, and colon if available, were examined from each animal. Twelve images were captured from each section and IgA-positive cells were counted and expressed per 10,000 μm(2). TLR5 genotypes for the G22A and C100T genes were determined by polymerase chain reaction on blood samples. Numbers of IgA-positive cells in the duodenum and colon were slightly higher than those published previously for GSDs with or without chronic enteropathy (mean in the crypt area of the duodenum 52.6 ± 16.2; mean in the tip of the duodenal villus 51.12 ± 3.83; mean in the base of the duodenal villus 55.02 ± 3.3; mean in the crypt area of the colon 67.4 ± 4.3). There was no correlation between numbers of IgA-positive cells in duodenum or colon between dogs carrying the RA versus the RP alleles of TLR genes. Further studies are needed to assess the production of secretory IgA and its relationship to TLR5 genotypes.

Enhancement of Mucosal Immune Responses in Chickens by Oral Administration of Cysteamine

Somatostatin, a tetradecapeptide originally isolated from the hypothalamus, can exert an inhibitory effect on the secretion of growth hormone by the anterior pituitary gland. In addition to endocrine organs, somatostatin is also present in the digestive tract and immune organs. In lymphoid tissues, somatostatin appears to play a role in the modulation of the immune response. Cysteamine (CS) is a sulfhydryl reducing agent that is known as a depletory agent of somatostatin. To evaluate its effects on chicken mucosal immune responses, CS was administrated orally to 1-wk-old broilers (40 mg/kg) that were immunized orally with Newcastle disease attenuated vaccine (NDV). The number of IgA-positive cells and intestinal intraepithelial lymphocytes (iIEL) in duodenum and jejunum were examined at 3-, 5-, and 7-wk posttreatment and immunization. The number of somatostatin-positive cells and relative amounts of somatostatin mRNA were also examined in the duodenum. The number of somatostatin-positive cells in the duodenum was reduced (P < 0.05) after CS treatment. In broilers receiving CS and NDV treatment (CS+NDV) the level of IgA-positive cells and iIEL in the duodenum and jejunum was increased (P < 0.05) at 3 and 5 wk posttreatment. The expression of somatostatin mRNA increased (P < 0.05) compared with that of the control group at 5 wk after immunization in broilers receiving CS+NDV or NDV alone. The results suggest that CS can induce proliferation and differentiation of IgA-positive cells and iIEL in the intestinal mucosa of chickens by reducing the number of somatostatin-positive cells.

Secretory IgA and squamous epithelization in adenoids of children with otitis media with effusion: Fujihara K.; Fujihara T.; Yamanaka N.Acta Oto-Laryngol. Suppl. (JPN) (1996) -/523 (155–157)

Infection of adenoids has been considered a causative factor of otitis media with effusion (OME). We believe that the deterioration in the mucosal barrier of adenoids makes the adenoids vulnerable to bacterial infections, resulting OME. In this study, we evaluated secretory IgA, secretory component (SC) and reticular and squamous changes of ciliated epithelium in adenoids. Fourteen children with OME (OME(+)) and 6 children without any history of OME (OME(+)) were enrolled in the study. Immunohistological study on the adenoids using anti-IgA and anti-SC revealed no significant difference between two groups in the number of IgA positive cells in the subepithelial layer. However, the proportion of the secretory component-positive area in the epithelium was significantly lower in the OME(+) group (11.4 +/- 13.0%) than in the OME(+) group (20.7 +/- 9.6%) (Mann Whitney U test, p <0.05). Histological study using hematoxylin and eosin staining revealed that the area of squamous epithelization was bigger in the OME(+) group (32%) than in the OME(+) group (16%) (p <0.05). The number of ciliated cells was decreased in the OME(+) group (13%) compared with the OME(+) group (31%). The epithelium was thicker in the OME(+) group (101 mu m) than in the OME(+) than in the OME(+) group (79 mu mm) (p <0.05). These results suggest that the production of the secretory component is lower in the epithelium of adenoids in children with OME. Hyperplasia, squamous epithelization and decreased number of ciliated cells may also hamper mucosal clearance of the adenoids of children with OME. These findings suggest that the adenoids of children with OME are more susceptible to infections than adenoids of children without OME.

Cells containing IgA subclasses in bronchi of subjects with and without chronic obstructive lung disease.

Necropsy specimens were obtained from the lungs of 10 subjects who had no history of lung disease, 10 who had died with chronic bronchitis, and 10 with bronchiectasis. Tissue sections were stained for IgA1 or IgA2 using the immunoperoxidase technique, and the number of cells in the bronchi stained for these proteins was counted. The total number of IgA positive cells was increased in bronchitic and bronchiectatic lungs compared with those from control subjects. The number of IgA2 positive cells was similar in those with bronchitis and bronchiectasis and significantly higher than in controls. Similarly, cells containing IgA1 were increased in the lungs of subjects with chest disease but were higher in those with bronchitis than in those with bronchiectasis. The proportion of IgA2:total IgA containing cells was similar in sections from controls (mean (SD) 25 (5.0)%) and those with bronchiectasis (mean (SD) 24 (4)%), but lower in those with bronchitis (mean (SD) 19 (5.0)%). The results show that cells containing IgA1 predominate in the major bronchi but that the proportion of cells containing IgA2 is higher than in non-mucosal lymphoid tissues. Bronchitis and bronchiectasis are associated with greater numbers of cells producing IgA in the bronchi, and this is consistent with increased local production of IgA in the lung secretions of bronchitic subjects.