实用器官移植电子杂志 ›› 2013, Vol. 1 ›› Issue (2): 82-86.

• 论著 • 上一篇    下一篇

输注分子嵌合前体 T 细胞减低小鼠脾脏T 细胞对异基因小鼠 T 细胞的刺激反应

付蔚华,宋东旭,王振,何向辉,李卫东
  

  1. 天津医科大学总医院普通外科,天津 300052
  • 出版日期:2013-03-20 发布日期:2021-04-20
  • 基金资助:
    国家自然科学基金资助项目(81041118)

Molecular chimeric pre-T cells infusion reduced spleen T lymphocyte response to allogeneic T cells

FU Wei-hua,SONG Dong-xu,WANG Zhen,HE Xiang-hui,LI Wei-dong.
  

  1. Department of General Surgery,TianjinMedical University General Hospital,Tianjin 300052,China
  • Online:2013-03-20 Published:2021-04-20

摘要:

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。

关键词:

Abstract:

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.

Key words: