Ly49A 基因转染的淋巴细胞对H—2^d小鼠正常和肿瘤细胞杀伤活性的实验研究

目的观察Ly49A基因转染的C57BL/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C57BL/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C57BL/6小鼠的淋巴细胞24?h后Ly49A受体表达率为(46.67±0.35)%，空载体转染组为(18.73±0.85)%，未转染对照组为(19.60±0.27)％，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22％～25%)，对4T1乳腺癌细胞的杀伤活性无明显改变（P＞0.05）。结论转染Ly49A的C57BL/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。

In vitro study of the killing activities on H-2(d) murine normal and tumor cells by Ly49A gene transfected lymphocytes]

OBJECTIVE: To observe the killing activities of Ly49A gene transfected lymphocytes of C57BL/6 mice to normal and tumor cells of BALB/c mice. METHODS: pLXSN-Ly49A retrovirus vector was constructed and packaged with PA317 cell line. The lymphocytes of C57BL/6 mice were transfected by culture with virus producing PA317 cells. The Ly49A expression rate on the transfected lymphocytes was detected by flow cytometry and the killing activities of the transfected lymphocytes to normal and tumor cells of BALB/c mice were assayed by MTT method. RESULTS: The Ly49A expression rates of C57BL/6 mice lymphocytes transfected with pLXSN-Ly49A for 24 hours, of those transfected with pLXSN and nontransfected control were (46.67 +/- 0.35)%, (18.73 +/- 0.85)%, and (19.60 +/- 0.27)%, respectively. The killing activity of the transfected lymphocytes to 4T(1) tumor cells remained almost the same as that of the control (P > 0.05), but to normal fibroblasts decreased sharply (inhibiting rate 22% - 25%). CONCLUSION: The Ly49A transfected C57BL/6 mice lymphocytes could kill BALB/c mice tumor cells as effectively as the control did, but the activity decreased sharply to normal BALB/c mice cells, which would be instructive for resolving graft versus host disease after allogeneic bone marrow transplantation.