猕猴单倍体相合外周血造血干细胞移植后多种细胞因子mRNA表达的动态分析

Kinetic Study of Various Cytokine mRNA Expressions in Rhesus Treated with Haploidentical Peripheral Blood Stem Cell Transp-lantation

本研究分析猕猴单倍体相合造血干细胞移植前后外周血单个核细胞中细胞因子(TGF-β,IL-2,IL-6,IL-10,IFN-γ,TNF-α,FAS-L)mRNA的表达并探讨这些细胞因子在急性移植物抗宿主病(aGVHD)发生和病理变化中的作用,以寻找能够早期预测和鉴别诊断aGVHD的指标。5例猕猴经非清髓预处理后接受单倍体相合外周血造血干细胞移植;半定量逆转录聚合酶链反应法(RT-PCR)检测移植前后8种细胞因子mRNA的表达,动态观察这些细胞因子在移植后aGVHD中的表达情况。结果表明:5例猕猴均成功获得造血重建。1例移植排斥,长期无病存活;其余4例为混合嵌合和完全嵌合植入,1例低比例嵌合植入经第二次输注供者外周血造血干细胞后获得高比例嵌合植入;1例发生肠道Ⅲ度GVHD。移植前后体内Th1和Th2类细胞因子都有不同程度的变化,在aGVHD期间TGF-β呈下调表达,其余均为上调表达;植入比例减低的猕猴各细胞因子也下降至移植前水平。结论:TGF-β在aGVHD发生前的下降趋势可能是GVHD预测指标,并且能够作为肠道GVHD的鉴别诊断指标。

This study was aimed to analyze the mRNA expression of cytokines （TGF-β, IL-2, IL-6,IL-10,IFN-γ, TNF-α, FAS-L ）in five rhesus treated with haploidenfical peripheral blood stem cell transplantation after nonmyeloablative preparative regimens and to explore the role of these cytokines in the development and pathology of acute graft-versus-host-disease （aGVHD）. Five rhesus monkeys received nonmyeloablative haploidentical peripheral blood stem cells transplantation. Semi-quantitative reversed transcription polymerase chain reaction （RT-PCR）was used to analyze the kinetics of cytokine mRNA expression in the transplantation and aGVHD. The results showed that five rhesus monkeys acquired hematopoietic reconstitution successfully. The graft was rejected in one monkey which survived without disease, the other four achieved mixed chimerism and full donor chimerism. Chimerism of low centigrade in one monkey achieved high centigrade at 35 days after donor stem cell infusion. Intestinal aGVHD grade Ⅲ developed in one monkey. Cytokines of Thl and Th2 changed after transplantation. In period of aGVHD, expression of TGF-β decreased but all others increased in various levels. When donor chimerism decreased, the cytokines decreased accordingly. It is concluded that the decrease of TGF-β mRNA may be an indicator to predict aGVHD, and can be used as a differential diagnostic indicator for intestinal GVHD.