H-2K^b基因导入小鼠造血细胞的研究

Transduction of H -2K^b gene into alice hematopoietic cells

本研究应用分子克隆技术，用脂质体（lipofectin）将MHC－Ⅰ类基因（即小鼠H－2kb基因）导入包装细胞PA317。用培养的病毒上清进一步感染BALB／C小鼠的造血细胞，且形成了抗G418的CFU－GM。通过多聚酶链式反应（PCB）和反转录-多聚酶链式反应（RT－PCR），表明病毒上清感染后的小鼠造血细胞及其形成的CFU－GM中均整合了MHC-Ⅰ类基因，且已转录为mRNA。流式细胞仪（FACS）测定结果表明，感染后的小鼠造血细胞及其形成的CFU－GM中均有MHC－Ⅰ类抗原的表达。本实验结果表明MHC－Ⅰ类基因已成功地导入小鼠造血细胞中，且不影响其增殖和分化能力。

This study aimed at looking for the possibilitiy of reducing the risk of immunological rejection through transplantation tolerance by transduction of MHC gene. A retroviral vector encoding MHC Ⅰgene,was transducted into packaging cells line PA317 by lipofectin. The helnatopoietic cells of BALB/C mice (H - 2Kb negative)were infected by the ahave viral supernatant. And their G418 - resistant CFU - GM were cultured. The hematopoietic cells of infected mice and their G418 - resistant CFU - GM were demonstrat to be able to express the MHC -Ⅰ gene by PCR and RT- PCR techniqUe. FACS deteCted the expression of MHC - Ⅰ antigen on the hematopoietic cells of infected face and their CFU - GM. These results that MHC - Ⅰ gene can be transducted into mice hematopoietic cells suceessfully, meanwhile their ability of proliferation and differentiation was not effected. These results would not only make ready for the next advanced research on inducing transplantation tolerance, but also provide evidences for the prevention and ment of immunological rejection in clinical transplantation.