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[Objective] mycophenolate mofetil (mycophenolate mofetil, MMF) is a new immunosuppressive agent, has been widely used in clinical allogeneic bone marrow transplantation can significantly reduce acute and chronic graft-versus-host disease (graft, versus Host disease The incidence and severity of GVHD), thereby improving the success rate of allogeneic bone marrow transplantation. MMF in vivo deesterification, the formation has immunosuppressive activity of mycophenolic acid (mycophenolate acid, MPA), MPA efficient, selective, non-competitive, reversible phosphorylation of hypoxanthine dehydrogenase (IMPDH) inhibitor inhibited guanine nucleotide classic synthesis pathway, due to the high dependence of T, B lymphocytes of the the purine classic synthesis pathway, MPA exert strong selective immunosuppressive effects. Previous studies that the occurrence of GVHD is mature donor T cells to host major histocompatibility antigen (MHC) or minor histocompatibility antigen (MiHA) identified as a foreign body and induce an immune response. In recent years, with the dendritic cells (dendritic cell, DC) study the deepening found DC also plays an important role in transplantation immunology. Recent experiments have shown that the MPA in addition to affecting the T and B cell function, can also directly inhibit the immunological function of antigen-presenting cells, dendritic cells is widely recognized as the antigen-presenting ability turned piece mysterious' the big month Bu months sigh d BU boxing spoon side Covent strong professional antigen presenting cells (antigen presentation cell, APC), the ability of T cell activation than macrophages, B cells and other APC 100 times, and is the only in vivo or in vitro activation of naive T cells, start the initial immune response APC. To this end, we have studied the M differentiation of cultured mouse bone marrow-derived dendritic cells mature and immunological function, as well as its NF-KB p50 expression levels to further explore MPA / MMF prevention GVHD the immune inhibition mechanism. Method 1 reference Inaba method and to improve slightly to cultured mouse bone marrow-derived DC were divided into three groups, a control group, that cultivate only add the GM-CSF 10ng/m1 IL-4 Ing / ml ; two group for MPAO.ol group, i.e. In addition to the above-mentioned cytokines added 0.01, mol / L MPA solution treated; three groups MPAO.1, i.e. in addition to the above cytokines added 0.1 p mol / L of the M in a solution processing. 2 inverted microscope and observed daily cell growth and collection of cultured cell suspension was centrifuged dry naturally after enrichment smear and Wri ght staining radiography. 3 mouse DC cell surface phenotype by flow cytometry. Added fluorescently labeled monoclonal antibody respectively for Iah eo4o a, eoso eDs6. 4 using mouse the of the chicken ovalbumin blood protein (OVA) antigen-specific MHCH class molecular limiting T cell hybridomas, the detection of the three groups of DC antigen-presenting function. 5 allogeneic mixed lymphocyte reaction (allo - MLR) to detect DC allogeneic T cell proliferation ability. 6 DC before the end of the culture, the culture supernatant the detection DC secreted IL-12 levels by ELISA kit; and MLR join: H TdR ago, the culture supernatant were detected by IFN by ELIsA kit a Ah, IL-2, IL-4 and IL-10, etc. of Thl / ThZ cytokine level. Off Art VII, Yue Bu Temple sigh Shi the fist-bit paper 7 immunoblotting (Western blot) to detect DC nuclear NF-KB p50 protein expression levels. [Results] after 6 days of culture can be immature DC with a typical dendritic structure, control group DC DC than MPAO.01 group DC and MPAO.1 of groups with more coarse dendritic membrane surface like pseudopodia. Compared with the control group, treated DC CD40, CD80 and CD86 molecule expression were significantly down-regulated, while MPA had no significant effect on the expression of the MHCn molecules. 3 MPA dose-dependent inhibition of antigen-presenting function of DC, the MPAO.01 the group, MPAO.l group and the control group comparison, P are (0.05 4 with the DC and the increase in the proportion of T cells in the control group, MPAO.01 group gradually enhanced and MPAO.l group DC ability to stimulate T lymphocyte proliferation and the MPAO.ol the group, MPAO.1 group and the control group comparison, P lt; 0 .05 5 DC before the end of the culture, the culture supernatant was collected to detect workers L level of a 12, the control group of mice DC able to secrete significant levels of IL-12 (437.89 ± 1 7.99 pg / ml), while MPAO.01 group and MPAO.1 of the secretion of IL-12 levels significantly decreased, respectively, 392.33 Guests 25.79 pg / ml and 228.64 ± 17.99 pg / ml (p lt; 0.05) 6 in all. MLR added: IH a TdR ago, the culture supernatant were detected by IL-2, workers FN, workers L -4 and IL a 10, etc. of Thl / ThZ cell factor level, results of IL-2 and IFN a Y level significantly with the rise, while workers L-4 and IL-10 level significantly with the decline, each group Liang Liang comparison, P are lt; 0 .05. 7 MPA can inhibit DC nuclear the NF-KBp50 expression level, and this effect is enhanced with increasing doses of MPA Zhedong dagger, the gas month Bu month sighed side of the seven the fist-bit paper [Summary] using mice bone marrow hematopoietic stem cells capable of inducing a large number of DC, and the morphology, phenotype and to stimulate the same kinds of cell proliferation functions confirmed. MPA liquid was added, can be cultured in DC phenotype and immunological functions are quite stable not Mature DC. we found that the MPA treatment DC more low the costimulatory molecule CD4O, CdSO and CD86 expression than the control group, DC maturation flag Since these costimulatory molecules is also prompted MPA treatment can prevent further maturation of the DC's. M handling the DC and the control group compared to their antigen presenting function, and T cell stimulating proliferation ability was significantly decreased, and thus can not effectively activate antigen-specific T cells, the T cells in the anergic state. MPA treatment DC secretion than a 12 feature significantly weakened. IL-12-DC maturation process in the secretion of one kind of cell factor, it is able to promote Tho cell differentiation as Thl cells, thereby inducing Thl-type cell mediated immune reaction the MPA processing the DC significantly improves ThZ cell factor IL a 4 and work L-10 level, and significantly with reduced Thl cell factor IFN keep and work L-2 level. showed in vitro, M processing DC induced allogeneic T cells to differentiate to ThZ thus conducive to generating the same species incompetence of antigen-specific T cells in vitro DC joined MPA can come down the DC nuclear NF KB p50 expression levels, and this effect is enhanced with increasing doses of MPA. MPA DC in vitro into
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