Tagami, S
Tagami, S. contamination. Autoimmune gastritis is not induced in is usually a chronic pathogen of the human gastric mucosa (40), infecting approximately (+)-Alliin half the world’s populace (20). Only 10 to 15% of infected individuals develop disease, which may range from acute gastric inflammation (38, 39) to duodenal and gastric ulcers, gastric adenocarcinoma, and mucosal-associated lymphoid tissue (MALT) lymphoma (10, 24, 51). contamination may explain the failure of infected individuals to induce immunity to contamination in human subjects with early gastric autoimmunity, as indicated by the presence of parietal cell-specific antibodies, suggests that contamination with may affect the induction or maintenance of stomach-specific autoimmunity (54), possibly as a result of molecular mimicry resulting from epitopes that are common to the gastric mucosa and contamination of BALB/c mice. These studies were designed to address the role of CD25+ Tregs in the maintenance of and growth conditions. CS1 (52) and SS1 (33) were obtained from A. H. Mitchell at The University or college of New South Wales, Sydney, Australia, and were cultured as explained by Sakagami et al. (57) and Lee et al. (33), respectively. Preparation of and antigens. Bacteria were harvested from broth culture or agar plates in PBS and sonicated while on ice. The bacterial sonicate was stored at ?70C, and the protein concentration was determined by a Bradford protein assay (Bio-Rad Laboratories). Contamination of mice with and CS1 was scraped from plates into brain heart infusion (BHI) broth, washed, and resuspended in BHI broth to approximately (+)-Alliin 108 bacteria per 200 l. SS1 was produced in BHI broth, washed, and resuspended in PBS to approximately 109 bacteria per 200 l. Prior to infecting mice, bacteria were analyzed in wet mounts for motility and morphology, as well as by urease test (25) and by Gram stain. Mice were infected on days 1, 3, and 5 by oral gavage with 200 l of bacteria under light anesthesia. Viable counts of the SS1 inoculum were determined immediately after contamination of mice by culturing the bacteria on selective agar plates under microaerophilic conditions. Assessment of and colonization. Stomachs were removed from euthanized mice and opened along the greater curvature. Contents were scraped, and the belly was washed twice in PBS and sectioned in small strips along its length to include the greater curvature. The belly strips were either fixed in 10% (vol/vol) formalin in 0.1 M Na-phosphate buffer (10% NBF), pH 7.2, washed with PBS, and frozen for immunohistochemistry or fixed in 10% NBF, processed, and embedded in paraffin, or used to enumerate the bacterial weight. colonization of the gastric mucosa was analyzed by histology. Paraffin-embedded tissues were slice (4 m) and silver stained using the Warthin-Starry method (42) to visualize the bacteria. The number of bacteria within the crypts of the antrum and body regions of the belly was enumerated in sections, and colonization was graded using a scoring method previously explained (69). colonization was quantified by determining the number of CFU per gram of belly tissue. Stomach strips were weighed, homogenized in 5 ml PBS, and serially diluted in PBS. The Miles and Misra dilution technique was used to enumerate CFU within each dilution (43). Aliquots were plated on Glaxo selective product agar plates (33). Histological examination and grading of gastritis. Hematoxylin and eosin-stained, formalin-fixed paraffin-embedded sections were used to grade the inflammatory response, based on a previously explained method (68). The belly mucosa was divided into upper, mid-, and lower body Mouse monoclonal to CD235.TBR2 monoclonal reactes with CD235, Glycophorins A, which is major sialoglycoproteins of the human erythrocyte membrane. Glycophorins A is a transmembrane dimeric complex of 31 kDa with caboxyterminal ends extending into the cytoplasm of red cells. CD235 antigen is expressed on human red blood cells, normoblasts and erythroid precursor cells. It is also found on erythroid leukemias and some megakaryoblastic leukemias. This antobody is useful in studies of human erythroid-lineage cell development and antrum. Mild inflammation was defined as an influx of inflammatory cells in the basal zone of the mucosa, moderate explains inflammatory cells extending up to the mid-zone, and in severe inflammation the infiltrate is usually spread through the full thickness of the mucosa. Lymphoid follicles were defined as selections of lymphocytes forming a central cortex and an outer marginal zone. Focal inflammation was defined as small aggregates of inflammatory cells often around a small blood vessel; diffuse inflammation explains cells forming a band in the lamina propria. The following six-point level was used to define mononuclear cell infiltration: 1, moderate multifocal; 2, moderate common or moderate multifocal; 3, moderate common and moderate or severe multifocal; 4, moderate common; 5, moderate common and severe multifocal; 6, severe widespread. The first 2 mm of the upper body at the junction with (+)-Alliin the esophageal mucosa were disregarded, since there was a consistent follicle in this zone in all infected mice. All sections were graded blindly. Flow.