| ObjectiveAnterior chamber-associated immune deviation (ACAID) is a deviant immune response induced by antigen introduced into the AC, which is characterized by the antigen-specific suppression of classical Thl immune responses, such as delayed-type hypersensitivity (DTH) and complement-fixing antibodies,while reserving the generation of non-complement-fixing antibodies of the IgGl isotype. It is one of the characteristic immune privilege mechanisms of the cornea, which is the major factor of long-term surviving of corneal implantation.It can avoid autoimmune disease and herpes simplex interstitial keratitis.MSCs can exert profound immunosuppression both in vitro and in vivo by inhibiting the proliferation and function of a number of immune cell types. It also has the role of immunoregulation and immune tolerance. In the present study, we plan to investigate the immunoregulatory effect of MSCs on ACAID development in mice model and clarify its mechanism, to provide new theoretical basis for clinical treatment of corneal allograft rejection reaction and autoimmune uveitis.Methods1.The culture and amplification of hMSC.2.C57BL/6 mice were divided into 4 groups, ACAID group (Group A),ACAID+MSCs group (Group B),positive control group (Group C) and positive control+MSCs group (Group D).2μl OVA (50mg/ml) was injected into the anterior chamber of A and B groups. Group C and Group D received AC injection of PBS. Meanwhile,B and D group got MSCs infusion via the tail vein (1×106/0.2ml MSCs).7ds later, all the mice received subcutaneous injections of 100μl OVA (2.5mg/ml OVA emulsified 1:1 in complete Freuned adjuvant).3.14ds later, DTH response was measured to evaluate the development of ACAID.4.15ds later, the levels of IL-10,TGF-β,IFN-γwere measured in supernatants derived from 48 hours cultures of splenic lymphocytes and OVA, using ELISA kits.5.15ds later, Flow cytometry was used to assay the frequency of CD4+CD25+Foxp3+T cell and CD8+Foxp3+T cell in the spleen of C57BL/6. Results1. hMSCs were successfully cultured and amplified.2. Mice models of anterior chamber-associated immune deviation were established.3. A significantly decreased DTH response were observed in the Group A and B, compared with Group C. But there was no significant difference between Group C and Group D.4.ELISA showed that the level of IL-10 in Group B was higher than that in GroupA, GroupC and Group D (P<0.05).There was no statistical significance between Group C and Group D; The level of TGF-βin Group B was higher than that in Group A, GroupC and Group D (P<0.05), and TGF-βin Group D is higher than that in Group C(P<0.05); The level of IFN-γin Group B is lower than that in Group A, Group C and Group D(P<0.05), and IFN-γin Group D is lower than Group C (P<0.05).5.A high level of CD4+CD25+Foxp3+T cell and CD8+Foxp3+T cell were observed in GroupB, compared with GroupA, GroupC and Group D (P<0.05). There was no statistical significance between Group C and Group D.ConclusionsTaken together, these results suggest that MSCs can suppress the antigen-specific DTH responses of ACAID, and promoting the maintainance of the immune deviation state effectivly, through the inhibition of pathogenic T-cell responses, modulation of the balance of Th1/Th2 and increasing the proportion of regulatory T cells. MSCs can protect the eye ball from inflammation injury caused by immune response, which provides new theoretical basis for the therapy of rejection after corneal transplantation and uveitis. |
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