Preliminary Study On Immunoregulatory Target Cells Of Fomes Officinals Polysaccharide

Objective:To assess the immune activity of polysaccharide from Fomes Officinals(FOPS,FOPS-a,FOPS-b)in vitro in order to explore the immunoregulative target cells of FOPS,FOPS-a,FOPS-b and preliminary study of their receptor or immunoreactive sites.Methond:1)The effects of FOPS,FOPS-a and FOPS-b on the activation and proliferation of macrophages were observed by cell proliferation and flow cytometry,and the effect of FOPS,FOPS-a and FOPS-b on the secretion of TNF-? and IL-1? and other inflammatory molecules measured by ELISA methods.2)CCK-8,flow cytometry and ELISA were used to observe the effects of FOPS,FOPS-a and FOPS-b on lymphocyte activation,proliferation and secretion of cytokines.3)The effect of anti-TLR2,anti-TLR4 and anti-CR3 on the secretion of TNF-? and IL-1? in macrophages by antibody blocking method.Results:1)The results of cell proliferation test showed that FOPS,FOPS-a and FOPS-b could significantly increase the proliferation and activation of macrophages,FOPS,FOPS-a and FOPS-b increased the levels of cytokines TNF-? and IL-1?,the results showed that macrophages might be one of the immunoregulatory target cells of FOPS,FOPS-a and FOPS-b.2)FOPS,FOPS-a and FOPS-b could significantly increase the proliferation activity of B lymphocytes induced by LPS,FOPS,FOPS-a and FOPS-b can promote the secretion of IL-2,IL-4,whereas it can promote immunoglobulin IgG of spleen lymphocytes,which suggested that B cells may be one of the immunoregulatory target cells of FOPS,FOPS-a and FOPS-b.3)Anti-TLR2 and Anti-TLR4 could inhibit the secretion of TNF-? and IL-1? in macrophages induced by LPS and FOPS,FOPS-a and FOPS-b,the results suggest that TLR2 and TLR4 may be the main action sites of FOPS,FOPS-a and FOPS-b on macrophages.4)Anti-TLR4 has a significant inhibitory effect on the proliferation activity of spleen lymphocytes induced by LPS and FOPS,FOPS-a and FOPS-b,indicating that TLR4 may be the action site of FOPS,FOPS-a and FOPS-b in B lymphocytes.Conclusion:1)Macrophages and B lymphocytes may be the main immunomodulatory target cells of FOPS,FOPS-a and FOPS-b.2)FOPS,FOPS-a and FOPS-b can promote the immune activity of RAW264.7 macrophages and spleen lymphocytes,and FOPS-a has the strongest immunomodulatory activity.3)TLR2 receptor and TLR4 receptor may be the main action sites of FOPS,FOPS-a and FOPS-b on macrophages.TLR4 receptor may be the main action sites of FOPS,FOPS-a and FOPS-b in B lymphocytes.