采用RT-PCR方法合成小鼠MHCⅡ类分子Ⅰ-Ak基因α和β链cDNA,插入逆转录病毒载体pLSXN,构建Ⅰ-Ak α和Ⅰ-Ak β表达载体,采用脂质体介导的重组质粒转移方法将Ⅰ-Ak α和Ⅰ-Ak β基因导入EL4小鼠淋巴瘤细胞和P815小鼠肥大细胞瘤细胞,经流式细胞仪检测在细胞表面有Ⅰ-Ak表达.将以上两种细胞注射到同源小鼠C57BL/6(H-2d)皮下,观察到肿瘤产生后又消退,证明在肿瘤细胞中单独导入同种异型MHCⅡ类分子基因也能激活肿瘤的细胞免疫,为进一步开展肿瘤的基因治疗奠定了基础.
In order to search the influence of Ⅰ-Akαβ gene transfer on the growth of the transferred mouse tumor cells in vivo, the α and β chain cDNA were synthesized, and inserted into a retroviral vector, to construct two Ⅰ-Ak expressing recombinant plasmids pLSXN-Akα and pLSXN-Akβ. By means of a liposome-mediated gene transfer procedure, these two recombinant plasmids together were introduced into mouse lymphoma EL4 cells and mastocytoma P815 cells. The expression of the Ⅰ-Akαβ protein on the surface of the transconducted tumor cells was tested by FACS using Anti-Ak-FITC. Afterwards these tumor cells were injected subcutaneously into autologous mice C57BL/6(H-2b) and DBA/2(H-2d) respectively. It was observed that tumors developed at the beginning within the mouse bodies, and disappeared after a few weeks with injection. While the nontransferred tumor cells grew continuously. These results show that the tumor immune response can be stimulated by the tumor cells transferred with allogenic MHC class Ⅱ gene, in the absence of B7 co-stimulatory signals.
丁广治,田军,刘元林,章扬培,毛宁.Ⅰ-Ak基因克隆转导及对肿瘤细胞成瘤的影响[J].生物化学与生物物理进展,1998,25(1):60-63
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