Primary Study Of The Role Of CD4 +~Foxp3~+Treg Cell In The Infection Of Helicobacter Pylori

Objectives: Construct the C57BL/6 mice model infected with H.pylori (Helicobacter pylori), comprehesive analysis the numbers of CD4~+Foxp3~+Treg cell and the level of related cytokines to evaluate the role of CD4~+Foxp3~+Treg cells in the infection of H. pylori.Methods:To construct the C57BL/6 mice animal model infected with H. pylori; Analysis the percentages of CD4~+Foxp3~+Treg cells in LPLs of gastric tissue and spleen lymphcyte by Flow Cytometry; PBS, supernatant of H. pylori by supersonic and PMA+Ionomycin+BFA were used as antigens to stimulate the cultured spleen lymphocyte of treatment group for 15 h, analysis the percentages of CD4~+Foxp3~+Treg cells by Flow Cytometry; The mRNA expression of Foxp3, IL-10 and TGF-β1 were detected by RT-PCR; ELISA were used to detect the secretion of IL-10, IFN-γand TGF-β1 in supernatant of gastric tissue homogenate, in serum and in supernatant of the spleen lymphocyte cultured and stimulated by antigens; Numbers and distribution of Foxp3~+ Treg cells in the gastric tissue was detected by Immunohistochemistry method; we used Hematoxylin and Eosin stain to observe the inflammation of mice and the therapeutic effect.Results:Percentages of CD4~+Foxp3~+Treg cell in gastric tissue LPLs and spleen lymphocyte of infection group separately were 1.22±0.25%, 2.97±0.16%, It was higher than that of PBS group and treatment group, P<0.01, there was statistical significance; Percentages of CD4~+Foxp3~+Treg cell in cultured spleen lymphocyte stimulated by supernatant of H. pylori supersonic was 2.18±0.05%, it was higher than that of PBS control group, but was lower than that of positive control group, P<0.01, there was statistical significance; A higher expression of Foxp3 mRNA in gastric tissue of infection group was detected, but no expression were observed in PBS group and treatment group; the secretion of IL-10 in cultured spleen lymphocyte stimulated by supernatant of H. pylori supersonic was 70.17±17.15(pg/mg), it was higher than that of PBS group and treatment group, P<0.01, there was statistical significance, and TGF-β1 was 1234.83±126.92(pg/mg), it was higher than that of PBS group, P<0.05, there was statistical significance; Number of Foxp3~+Treg cell in gastric tissue of infection group was higher than that of PBS group and treatment group, P<0.05, there was statistical significance.Conclusions:(1) The numbers and percentage of CD4~+Foxp3~+Treg cell increase in the infection sites and spleen after infected with H. pylori, then decrease follow the eradication of H. pylori, CD4~+Foxp3~+Treg cell and H. pyori infection is closely related.(2) There are higher expression of Foxp3 mRNA in the gastric tissue of H. pylori infection.(3) Supernatant of H. pylori by supersonic could effectively induce the CD4~+ T cell in spleen lymphocyte to differentiate into Treg cell, promote the secretion of IL-10 and TGF-β1.