南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (04): 394-397.

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含hTERT片段的重组逆转录病毒感染对树突状细胞功能的影响

胡贵方； 孙莉莎； 金宏； 欧程山； 蒋毅萍； 庞建新；   

  1. 南方医科大学流行病教研室； 南方医科大学药理学教研室； 南方医科大学毒理学教研室； 南方医科大学药理学教研室 广东 广州 510515； 广东 广州 510515；
  • 出版日期:2006-04-20 发布日期:2006-04-20
  • 基金资助:
    国家自然科学基金(30000208);广东省自然科学基金(32876)~~

Functional changes of dendritic cells after infection by recombinant retrovirus carrying human telomerase reverse transcriptase gene fragment

HU Gui-fang, SUN Li-sha, JIN Hong, OU Cheng-shan, JIANG Yi-ping, PANG Jian-xin Department of Epidemiology, Department of Pharmacology, Department of Toxicology, Southern Medical University, Guangzhou 510515, China   

  1. 南方医科大学流行病教研室； 南方医科大学药理学教研室； 南方医科大学毒理学教研室； 南方医科大学药理学教研室 广东 广州 510515； 广东 广州 510515；
  • Online:2006-04-20 Published:2006-04-20

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摘要： 目的观察含人端粒酶逆转录酶(hTERT)片段的重组逆转录病毒感染对树突状细胞(DCs)功能的影响。方法 ELISA试剂盒检测DCs培养液中IL-12水平;混合白细胞(MLR)反应检测含hTERT片段的重组逆转录病毒感染的 DCs(hTERT-DCs)和未感染的DCs(N-DCs)刺激同种异体淋巴细胞增殖能力;流式细胞术检测DCs表面分子CD80、 CD83、CD86和HLA-DR的变化;CytoTox 96非放射性细胞毒性检测试剂盒检测细胞毒性T淋巴细胞(CTL)反应。结果 hTERT-DCs和N-DCs在分泌IL-12的水平、刺激同种异体淋巴细胞增殖的能力方面无明显差异;hTERT-DCs的 CD83表达水平低于N-DCs,同时,hTERT-DCs激发的CTL对端粒酶阳性的靶细胞杀伤率明显高于端粒酶阴性的靶细胞(P<0．05)。结论 hTERT-DCs尽管有可能阻止DCs自身的成熟,但在活化淋巴细胞、刺激淋巴细胞分化增殖的能力方面并没发生明显改变,并且还能激发hTERT特异性CTL。 

Abstract: Objective To observe the functional changes of dendritic cells (DCs) after infection by recombinant retrovirus carrying human telomerase reverse transcriptase (hTERT) gene fragment. Methods Interleukin-12 (IL-12) levels in DC culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA). The abilities of DCs infected with recombinant retrovirus carrying hTERT gene (hTERT-DCs) and non-infected DCs (N-DCs) to stimulate allogeneic lymphocyte proliferation were evaluated with mixed leukocytes reaction (MLR), and the surface markers of DCs including CD80, CD83, CD86 and HLA-DR were detected by flow cytometry. Cytotoxic T lymphocyte (CTL) assay was performed with CytoTox 96 non-radioactive cytoxicity assay. Results Compared with N-DCs, hTERT-DCs showed no significant changes in IL-12 secretion and capacity to stimulate allogeneic lymphocytes reaction, but had significantly lower CD83 expression. Specific CTLs induced by hTERT-DCs resulted in higher cytotoxicity against telomerase-positive target cells than that against the negative target cells. Conclusions Infection with the recombinant retrovirus carrying hTERT fragment may jeopardize the maturation of DCs, which, however, still retain their capacity to activate and stimulate lymphocyte proliferation and to prime autologous T lymphocytes to generate specific CTL against hTERT. 

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