论文部分内容阅读
目的 观察反义寡核苷酸封闭鼠淋巴细胞H-2Kd基因后,对其表达的影响及淋巴因子激活杀伤细胞(LAK)的活性改变,探讨淋巴细胞Ⅰ类主要组织相容性抗原(MHC-Ⅰ)在肿瘤免疫中的作用。方法 将人工合成的H-2Kd起始区反义寡核苷酸作用于小鼠脾LAK,流式细胞仪检测作用前后H-2Kd的表达率。噻唑蓝法检测H-2Kd表达降低的LAK对肿瘤杀伤活性。结果 反义寡核苷酸(15μmoL/l)组H-2Kd蛋白的表达率由90.1％±4.4％下降至79.8％±2.5％(P<0.01),H-2Kd降低的LAK杀伤活性明显降低,对K562的杀伤活性由82.3％±3.1％降到60.2％±6.7％(P<0.01),对H22细胞株的杀伤活性由67.4％±2.8％降到53.2％±8.1％(P<0.01)。结论 反义寡核苷酸能够抑制小鼠LAK表面H-2Kd的表达,但不影响LAK增殖活性,因而可导致鼠LAK对同种及异种肿瘤细胞的杀伤活性降低。
Objective To investigate the effects of antisense oligonucleotide on the expression of H-2Kd in murine lymphocytes and the changes of activity of lymphokine-activated killer (LAK) cells in order to explore the role of MHC- Ⅰ) in tumor immunity. Methods The synthetic H-2Kd antisense oligonucleotide was used to detect the expression of H-2Kd in spleen of mice. The expression of H-2Kd was detected by flow cytometry. The cytotoxic activity of LAK with reduced H-2Kd expression was detected by the thiazolyl blue assay. Results The expression of H-2Kd decreased from 90.1% ± 4.4% to 79.8% ± 2.5% (P <0.01) in antisense oligonucleotide group (15μmol / L) The killing activity to K562 decreased from 82.3% ± 3.1% to 60.2% ± 6.7% (P <0.01), and the killing activity to H22 cell line decreased from 67.4% ± 2.8% to 53.2% ± 8.1% (P <0.01). CONCLUSION: Antisense oligonucleotide can inhibit the expression of H-2Kd on the surface of mouse LAK, but it does not affect the activity of LAK. Therefore, antisense oligonucleotide can reduce the killing activity of LAK on allogeneic and xenograft tumor cells.