Objective To construct the mouse chimeric pre-T cells with major histocompatibilitycomplex-Ⅰ（MHC-Ⅰ）gene，and explore the potential of chimeric pre-T cells to alleviate spleen T lymphocyteresponse to allogeneic mouse T cells. Methods BALB/c mouse pre-T cells were cultured. Eukaryotic expressionvectors of C57BL/6 mouse MHC-Ⅰ（pIRES-H-2Db and pIRES-H-2Kb）wereconstructed to transfect BALB/c pre-Tcells. Then the molecular chimeric cells were transfused back to BALB/c mouse. After 7 days，the effect of molecularchimeric cells on spleen T lymphocyte response to allogeneic T cells was observed through mixed lymphocyte culture

（MLC）. Stimulation index（SI）was calculated. Results BALB/c mouse pre-T cells were successfully separatedand cultured. Flow cytometry analysis indicated that H-2Db and H-2Kb protein expression were（14.90±0.56）%and（14.20±0.63）% in pre-T cells group. The result of MLC demonstrated that the SI of T lymphocyte weresignificantly decreased in molecular chimeric cells group（0.764±0.074）compared with empty vector group（0.983±0.081）and non-transfection group（0.994±0.142，both P＜0.05）. There were significances in SI betweenmolecular chimeric cells group and pIRES-H-2Db group（0.859±0.085），pIRES-H-2Kb group（0.860±0.097，both P＜0.05）. Conclusion The molecular chimeric pre-T cells infusion could alleviate spleen T lymphocyte responseto allogeneic T cells.

Pre-T cell, Major histocompatibility complex-Ⅰ gene, Molecular chimerism

目的 构建分子嵌合主要组织相容性复合体（MHC）-Ⅰ基因小鼠前体 T 细胞，并探讨其诱导脾脏 T 细胞减低对异基因小鼠 T 细胞反应的可行性。方法 体外分离培养 BALB/c 小鼠前体 T 细胞，构建携带 C57BL/6 小鼠 MHC-Ⅰ基因（H-2Db 和 H-2Kb）真核表达载体 pIRES-H-2Db 和 pIRES-H-2Kb，分别转染 BALB/c 小鼠前体 T 细胞，构建分子嵌合前体 T 细胞。将分子嵌合前体 T 细胞回输BALB/c 小鼠后 7 天，获取脾脏 T 淋巴细胞，与 C57BL/6 小鼠 T 细胞进行混合淋巴细胞培养，观测刺激指数（SI）。结果 成功体外培养BALB/c 小鼠前体 T 细胞，体外转染 C57BL/6 小鼠 H-2Db 和 H-2Kb 基因至 BALB/c 小鼠前体 T 细胞，H-2Db 和 H-2Kb 蛋白表达率分别可达（14.90±0.56）% 和（14.20±0.63）%。单向混合淋巴细胞培养显示，输注分子嵌合前体 T 细胞的 BALB/c 小鼠脾脏 T 细胞对 C57BL/6 小鼠 T 细胞 SI，在 pIRES-H-2Db 和pIRES-H-2Kb 转染的小鼠前体 T 细胞共注射组为（0.764±0.074），比空质粒组（0.983±0.081）和未转染组（0.994±0.142）明显下降（均 P ＜0.05）。共注射组 SI 值分别与转染质粒 pIRES-H-2Db 组 SI 值（0.859±0.085）和转染质粒 pIRES-H-2Kb 组 SI 值（0.860±0.097）相比，差异均有统计学意义（均 P ＜0.05）。结论输注分子嵌合 MHC-Ⅰ基因前体 T 细胞的小鼠脾脏 T 细胞对异基因小鼠 T 细胞刺激反应明显减低。

前体 T 细胞 , 主要组织相容性复合体 -Ⅰ基因 , 分子嵌合