混合胸腺裸小鼠移植模型的建立及重建T细胞功能的体外研究

Functions of reconstituted T cells in vitro by grafting with mixed fetal thymus in nude mice

目的 建立混合胸腺移植模型 ,并在体外观察重建T细胞的功能状况。方法 首先建立混合胸腺裸小鼠左肾包膜下移植模型 ,流式细胞仪检测受体动物外周血中T细胞的重建情况 ,并利用刀豆素A(concanavalinA ,ConA)增殖实验和混合淋巴细胞培养检测其体外功能状态。结果 混合胸腺移植 3个月后 ,移植的胸腺组织在裸小鼠肾包膜下能够正常发育 ,并有效重建了受体外周血中的CD4+ T和CD8+ T细胞。体外功能检测显示 ,受体的脾细胞对ConA的刺激有良好的反应性 ,并对供体胸腺来源的F3Q44大鼠抗原刺激表现为特异性的低反应。结论 混合胸腺移植不仅在数量上可重建受体裸小鼠外周血中CD4+ T和CD8+ T细胞 。

Objective To establish the model of mixed thymus transplantation by grafting with fetal F344 rat thymus and BALB/c mouse thymus in nude mice and to observe the functions of the reconstituted T cells in the recipients. Methods Transplantation model was established by implantation of the mixed fetal thymic tissue under the kidney capsule in nude mice. The reconstitution of T cell subsets in peripheral blood of the recipients was detected by flow cytometry. The in vitro functional status of the reconstituted T cells in the nude mice and mouse splenocytes was stimulated with Con A mitogens, allogeneic and xenogeneic splenocytes. Results At 3 months after mixed thymus transplantation, efficient reconstitution of mouse CD4 +T cells and CD8 +T cells in the peripheral blood was observed. The levels of mouse CD4 +T cells and CD8+T cells in peripheral white blood cells were similar to those of the normal euthymic mice. Con A stimulation and mixed lymphocyte reaction assay revealed that mouse lymphocytes showed a strong response to either the stimulation of allogeneic (C57BL/6 mouse) cells or Con A mitogens, and were tolerant for donor F344 rat antigens. Conclusion Mixed thymus transplantation can support the reconstitution of CD4 +T cells and CD8 +T cells in nude mice. Furthermore, the function of reconstituted T cells is normal in vitro .