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背景与目的:树突状细胞(dendritic cell,DC)作为一种专职的抗原提呈细胞,能激活并刺激初始(na-ive)T淋巴细胞抗击外来微生物及肿瘤,在诱导免疫应答中起着特殊的作用。本研究通过体外检测不同抗原负载的DC刺激小鼠TC的增殖、诱导肿瘤特异性CTL分泌INF-γ的量以及对A20细胞的杀伤实验,探讨DC体外活化TC的效应。方法:混和淋巴细胞反应(MLR)用MTT法检测不同抗原负载后的DC体外刺激同种异体淋巴细胞的增殖率;LDH法检测不同抗原负载后的DC诱导同种异体CTL的特异性杀伤活性;ELISPOT法检测活化的TC分泌INF-γ的能力。结果:在DC∶TC=1∶10时,CPP-Id-DC-TC(320%±15%)的增殖率明显高于Id-DC-TC(57%±10%)(P<0.005);CPP-Id-DC-TC对A20细胞的杀伤率(58%±10%),明显高于其对B16细胞(8.2%±1.2%)(P<0.001)及Id-DC-TC对A20细胞的杀伤率(21.6%±8.6%)(P<0.005);CPP-Id-DC-TC组产生的斑点数(96±8)个比Id-DC-TC组(31±6)个及单纯TC组(9±2)个产生的斑点数明显增加,差异有显著性(P<0.01)。结论:CPP-Id体外冲击DC后,可以诱导CTL的增殖及杀伤活性增强。
BACKGROUND AND PURPOSE: Dendritic cells (DCs), as specialized antigen-presenting cells, activate and stimulate na [iota] ve T-lymphocytes against foreign microbes and tumors and play an important role in inducing immune responses Special effect In this study, in vitro detection of different antigen-loaded DC stimulation of mouse TC proliferation induced tumor-specific CTL secretion of INF-γ and A20 cell killing experiments to explore the effect of DC in vitro activation of TC. Methods: Mixed lymphocyte reaction (MLR) was used to detect the proliferation rate of allogeneic lymphocytes stimulated by DC in vitro with MTT assay. LDH was used to detect the specific cytotoxic activity of allogeneic CTL induced by DCs. The ELISPOT assay was used to examine the ability of activated TCs to secrete INF-γ. Results: The proliferation rate of CPP-Id-DC-TC (320% ± 15%) was significantly higher than that of Id-DC-TC (57% ± 10% The killing rate of A20 cells by CPP-Id-DC-TC (58% ± 10%) was significantly higher than that of B16 cells (8.2% ± 1.2% (96 ± 8) in the CPP-Id-DC-TC group (31 ± 6) compared with the pure TC group (21 ± 8.6%) in the Id-DC-TC group (9 ± 2) spots produced a significant increase in the number of spots, the difference was significant (P <0.01). Conclusion: CPP-Id can enhance the proliferation and cytotoxicity of CTL induced by DC in vitro.