Exploring The Effect And Mechanism Of Acquired Regulatory "Th3" Cells In IgA Nephropathy

Objective:Exploring the effect and mechanism of acquired regulatory "Th3" cells in IgAnephropathy, in order to find new train of thought and theoretical basis for IgA nep--hropathy in clinical treatment.Methods:By cell experiments in vitro,the uniform suspension and equal volume Dakiki Blymphocytes was inoculated in culture plate,a negative control group (without anystimulating factors),the experimental group (with TGF-beta1in different concentra--tion respectively:5ng/ml,10ng/ml,15ng/ml,30ng/ml)and the positive control group(with IL-4,10ng/ml). All groups were cultured for3days and7days respectively.Using the double antibody enzyme-linked immuno sorbent assay (ELISA) to detectthe immunoglobulin IgA1concentration, the degree of abnormal IgA1glycosylation,the degree of IgA1saliva acid of the supernatant fluid for all groups; Using real-timefluorescent quantitative PCR technology to detect the mRNA gene expresssion levelsof C1β3Gal-T galactosyltransferase and Comsc molecular chaperone in all groups Blymphocytes which were cultured for12hours and72hours respectively in differentconcentration of TGF-beta1and IL-4factor stimulating.Results:(1)The IgA1concentration detection of the supernatant fluid: ELISA detectionresults showed that transforming growth factor TGF-beta1adjusted B lymphocyte tosecrete immunoglobulin IgA1by a two-way: positive correlation at lowconcentrations (TGF-beta1,0-10ng/ml), but negative correlation at high concentration(TGF-beta1,10-30ng/ml), blank group versus all experimental groups and IL-4group in3days or7days, all P <0.05, it has statistical significance.(2)The detection of degree of abnormal IgA1glycosylation: Certainconcentration of cytokine TGF-β1and IL-4can promote B lymphocyte to secretethe producton of abnormal glycosylated IgA1, blank group versus all experimentalgroups and IL-4group in3days or7days, all P <0.05, it has statistical significance. (3)The detection of mRNA gene expression of C1β3Gal-T galactosyltransferaseand Cosmc molecular chaperone: Real time-PCR detection results showed thatCertain concentration of cytokine TGF-beta1downregulated the two gene expressionof C1β3Gal-T galactosyltransferase and Cosmc molecular chaperone, the geneexpression relative quantitative of C1β3Gal-T galactosyltransferase and Cosmcmolecular chaperone for blank group VS all experimental groups and IL-4group, allP <0.05, there is statistical significance, however, all experimental groups and IL-4group by pairwise comparison, all P>0.05, it has no statistical significance.Conclusions:Certain concentration of Th3type cytokine TGF-β1could promote the secretionof IgA1by B lymphocyte and influenced the level of IgA1glycosylation. Themechanism may be that TGF-β1made level of IgA1glycosylation changed bydown-regulating the two gene expression of C1β3Gal-T galactosyltransferase andComsc molecular chaperone, and thus involved in the immune pathogenesis of IgAnephropathy.