MSCs Alleviate PSS Patients Complicated With ITP By Regulating The Expression Of Fc? R?b On Monocytes And The Role Of CD177 In PSS Patients With Refractory ITP

Objective: To observe the therapeutic effect of MSCs transplantation in p SS-ITP patient and explore the mechanism,so as to advance the MSCs therapy in future clinical applications.Methods: The expression of Fc?RI,II,IIb and III on monocytes of p SS-ITP,p SS patients,healthy controls and the expression of Fc?Rs before and after MSCs treatment in p SS-ITP patients were detected by flow cytometry;The changes of platelet phagocytosis ability of monocytes/macrophages before and after MSCs treatment in p SS-ITP patients were detected by flow cytometry and observed by laser confocal microscopy;ELISA was used to detect the levels of cytokines in the co culture supernatant of MSCs and p SS-ITP monocytes,si IL-10 MSCs were co-cultured with monocytes from p SS-ITP patients to observe whether the regulatory effect could be reversed.Results: We found that the expression level of Fc?R IIb/CD32 b on monocytes of p SSITP patients was significantly lower than that of HC and p SS,which was negatively correlated with the level of anti-SSB antibody,ESSDAI,while positively correlated with platelet count;After MSCs treatment,the expression level of Fc?R IIb/CD32 b on monocytes of p SS-ITP patients was up-regulated,but there was no significant difference of Fc?RI/CD64,Fc?RII/CD32 and Fc?RIII/CD16 before and after MSCs treatment.The concentration of IL-10 increased significantly in the co culturesupernatant of MSCs and p SS-ITP monocytes.Besides that,the phagocytosis of monocytes/macrophages in patients with p SS-ITP was significantly decreased after MSCs treatment;si IL-10 MSCs could not up regulate the expression of Fc?RIIb/CD32 b on monocytes in patients with p SS-ITP.Conclusion:(1)MSCs had good response rate and is well tolerated in the treatment of p SS-ITP;(2)The phagocytic activity of monocytes/macrophages greatly decreased after MSCs treatment;(3)MSCs up-regulate the expression of Fc?RIIb/CD32 b through IL-10.Objective: To detect the expression of Fc?RI,IIb and III in spleen macrophages of NOD-ITP mice treated with MSCs,and to explore the mechanism of MSCs on Fc?Rs system.Methods: The mice were randomly divided into: 1)NOD group;2)NOD-ITP group;3)si NC MSCs treated group;4).si IL-10 MSCs treated group.The expression of Fc?RI,IIb and III in splenic macrophages of each group was detected by flow cytometry and the platelet clearance in vivo was tested by phagocytosis assays.Results: Compared with NOD-ITP and si IL-10 MSCs treatment group,the platelet count and plasma IL-10 level of si NC MSCs treatment group were significantly higher;Compared with NOD-ITP group,the expression of Fc?RIIb but not Fc?RI and Fc?RIII in spleen macrophages was significantly up-regulated in si NC MSCs transplantation group;while the expression of Fc?RI,IIb,III on the surface of splenic macrophages in si IL-10 MSCs transplantation group had no significant difference;In vitro phagocytosis assays,it was showed that the clearance coefficient of NOD-ITP group and si IL-10 MSCs group was significantly higher than that of si NC MSCs group.Conclusion: MSCs decreased platelet clearance and up regulated the expression of Fc?R IIb in splenic macrophage of NOD-ITP mice through IL-10.Objective: To determine whether the low expression of CD177 affects the directional differentiation of hematopoietic stem cells into megakaryocytes,and to explore the expression and significance of CD177 in peripheral neutrophils of p SS-ITP.Methods: Bone marrow CD34+ cells from 3 healthy controls and 4 p SS-RITP patients were collected for transcriptome microarray analysis to screen differentially expressed genes.The differentially expressed genes of bone marrow CD34+ cells were verified by RT-PCR from healthy controls and p SS-RITP patients.In vitro,CD177 was silenced in hematopoietic stem cells to detect its effect on the proliferation of hematopoietic stem cells and the directional differentiation of megakaryocyte.Forty patients with newly diagnosed of p SS-ITP were followed up 48 weeks,and then they are divided into p SS-RITP and p SS-NRITP according to the treatment and response to drugs.RTPCR was used to detect the expression of CD177 m RNA in peripheral blood neutrophils at baseline,12 W,24W and 48 W.Results: The results showed that the expression of CD177 in bone marrow CD34+ cells of p SS-RITP patients were significantly decreased;In vitro,silencing of CD177 did not inhibit the proliferation of CD34+cells,affect the directional differentiation of CD34+cells into megakaryocytes and the formation of polyploidy;Besides that it had no effect on the adhesion and migration of megakaryocytes,and could not inhibit the rearrangement of cytoskeleton microfilaments of megakaryocytes.The CD177 m RNA expression on neutrophils in peripheral blood of patients with p SS-RITP at 12 W,24W,48 W was lower than that of p SS-NRITP patients.Conclusion:(1)Low expression of CD177 could not affect the directional differentiation of CD34 + cells into megakaryocytes;(2)The low expression of CD177 m RNA in peripheral blood neutrophils of newly diagnosed of p SS-ITP patients at 12 W may be a predictor of the development of p SS-RITP.