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目的 观察Ly49A基因转染的C5 7BL/ 6小鼠的淋巴细胞对BALB/c小鼠正常成纤维细胞和 4T1乳腺癌细胞杀伤能力的变化与差异。方法 构建逆转录病毒表达载体pLXSN Ly49A ,经由PA317包装细胞包装后转染C5 7BL/ 6小鼠淋巴细胞。流式细胞仪检测Ly49A受体在转染后淋巴细胞上的表达率。MTT法检测转染后淋巴细胞对BALB/c小鼠正常成纤维细胞和 4T1乳腺癌细胞的杀伤活性 ,以空载体转染和未转染的淋巴细胞作对照。结果 Ly49A基因转染的C5 7BL/ 6小鼠的淋巴细胞 2 4h后Ly49A受体表达率为 (4 6 .6 7± 0 .35 ) % ,空载体转染组为 (18.73± 0 .85 ) % ,未转染对照组为 (19.6 0± 0 .2 7) % ,其对BALB/c小鼠正常成纤维细胞的杀伤活性明显降低 (抑制率 2 2 %～ 2 5 % ) ,对 4T1乳腺癌细胞的杀伤活性无明显改变 (P >0 .0 5 )。结论 转染Ly49A的C5 7BL/ 6小鼠淋巴细胞对BALB/c小鼠正常细胞的杀伤作用明显降低 ,但仍保留了对肿瘤细胞的杀伤活性 ,为解决异基因骨髓移植后移植物抗宿主病提供了实验依据
Objective To observe the changes and difference of the killing ability of Lymphocytes in C57BL/6 mice transfected with Ly49A gene to BALB/c mice normal fibroblasts and 4T1 breast cancer cells. Methods The retroviral expression vector pLXSN Ly49A was constructed and transfected into C57BL/6 mouse lymphocytes by packaging PA317 packaging cells. The expression rate of Ly49A receptor on lymphocytes after transfection was detected by flow cytometry. MTT assay was used to detect the cytotoxicity of transfected lymphocytes against normal fibroblasts and 4T1 breast cancer cells in BALB/c mice. The control cells were transfected with empty vector and untransfected lymphocytes. Results Ly49A receptor expression rate was (46.67±0.35)% in Ly49A-transfected C57BL/6 mice after 24 hours, and (18.73±0.85) in empty vector transfection group. %, the untransfected control group was (19.6 0 ± 0.27%), its killing activity on normal fibroblasts in BALB/c mice was significantly reduced (inhibition rate of 2% to 25%), and to 4T1 mammary glands There was no significant change in the killing activity of cancer cells (P > 0.05). Conclusion The killing effect of Ly49A-transfected C57BL/6 mouse lymphocytes on normal cells of BALB/c mice was significantly reduced, but the cytotoxicity to tumor cells was still retained. This study aimed to solve graft-versus-host disease after allogeneic bone marrow transplantation. Provide experimental basis