CD200/CD200R在异基因造血干细胞移植后的表达水平及意义

CD200/CD200R Expre ssion Levels and Its Significance after Allogeneic Hematopoietic Stem Cell Transplantation

背景:CD200与其受体CD200R均属于免疫球蛋白超家族成员,在体内起到传递免疫信号、参与免疫稳态的维持、调节自身免疫功能紊乱以及抑制巨噬细胞系统的功能。目的:探讨CD200/CD200R在移植物抗宿主病(GVHD)中的作用。方法:将入组患者分为4组:aGVHD组、non-aGVHD组、cGVHD组及non-cGVHD组。首先应用流式细胞术测定各组CD200及CD200R在CD19+细胞、CD3+细胞及树突状(DC)细胞表面表达水平的差异,随后通过实时荧光定量PCR测定各组CD200及CD200R的mRNA表达水平的差异,最后将各研究组外周血单个核细胞与不同浓度的抗-CD200R1抗体体外共培养48 h,采用ELISA方法测定培养上清IL-10水平,以验证CD200/CD200R的功能。结果:non-aGVHD组CD19+细胞表面CD200+表达水平高于aGVHD组,低于正常对照组。non-cGVHD组CD19+细胞表面CD200+表达水平高于正常对照组及cGVHD组。aGVHD组和cGVHD组CD200的mRNA的表达水平均低于正常对照组。aGVHD组CD200的mRNA表达水平低于non-aGVHD组,cGVHD组低于non-cGVHD组。各组CD200R的mRNA表达水平未见明显差异。抗-CD200R1抗体阻断CD200/CD200R通路后,细胞培养上清中IL-10浓度随抗-CD200R1抗体浓度的增加呈下降趋势。结论:aGVHD及cGVHD患者在CD19+CD200+细胞及mRNA表达水平均明显低于未发生GVHD的患者,阻断CD200/CD200R通路后IL-10分泌水平下降,提示CD200/CD200R可能与GVHD的发生有一定的相关性。

Background: CD200 and its receptor CD200 R are both type-1 membrane glycoproteins,w hich are members of the immunoglobulin superfamily( Ig SF). Besides the inhibitory effect on macrophages,CD200 / CD200 R also play an important role in regulating the regulatory T cells,allergicreaction,autoimmune diseases,allograft,neurological diseases and other autoimmune-related diseases,etc. Objective: To investigate the role of CD200 and its receptor in the graft versus host disease( GVHD). Methods: Experimental samples w ere divided into aGVHD group,non-aGVHD group,c GVHD group and non-c GVHD group,the healthy persons w ere used as normal controls. Firstly,the expression levels of CD200 and CD200 R on CD19+cell,CD3+cell and dendritic cell( CD19-CD14-CD1c+)surfaces in each group w ere detected by using flow cytometry,so as to determine w hether there w ere expression differences among each groups. Then,the mRNA levels of each groups w ere tested by using real-time quantitative polymerase chain reaction for finding the differences of mRNA expression level among each group. Finally,the peripheral blood mononuclear cells of the patients and healthy controls w ere co-cultured w ith anti-CD200R1 antibody for48 hours,and the interleukin-10 level in the co-culture system w as tested by using enzyme linked immunosorbent assay for verifying the function of CD200 / CD200 R. Results: The CD200 expression level on CD19+cell surface in the aGVHD group and non-aGVHD group w as both low er than that in healthy control group; that in the non-aGVHD group w as higher than that in the aGVHD group. The CD200 expression level on CD19+CD200+cells in the non-c GVHD group w ere higher than that in the c GVHD group and healthy control group. There w ere no significant differences of CD200 and CD200 R expression levels on CD3 cells and dendritic cells among all groups. The CD200 mRNA expression levels in the aGVHD group and c GVHD group w ere both low er than the healthy control group. The CD200 mRNA expression level w as low er in the aGVHD group than in the non-aGVHD group,and w as low er in the c GVHD group than in the non-c GVHD group. There w as no significant difference of the CD200 R mRNA expression level among all groups.After the peripheral blood mononuclear cells of the patients and healthy controls w ere co-cultured w ith anti- CD200R1 antibody for 48 hours,the interleukin- 10 concentration decreased w ith the increasing of anti-CD200R1 antibody concentrations in the co-culture system. Conclusion: The CD200 / CD200 R may play a role in the pathogenesis of GVHD after allo-HSCT.