Hu Gui-fang, Sun Li-sha, Jin Hong, Ou Cheng-shan, Jiang Yi-ping, Pang Jian-xin
Department of Epidemiology, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2006 Apr;26(4):394-7.
To observe the functional changes of dendritic cells (DCs) after infection by recombinant retrovirus carrying human telomerase reverse transcriptase (hTERT) gene fragment.
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.
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.
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.
观察携带人端粒酶逆转录酶(hTERT)基因片段的重组逆转录病毒感染后树突状细胞(DCs)的功能变化。
采用酶联免疫吸附测定(ELISA)法检测DC培养上清液中白细胞介素-12(IL-12)水平。用混合淋巴细胞反应(MLR)评估携带hTERT基因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的能力,并用流式细胞术检测DCs的表面标志物,包括CD80、CD83、CD86和HLA-DR。采用CytoTox 96非放射性细胞毒性测定法进行细胞毒性T淋巴细胞(CTL)测定。
与N-DCs相比,hTERT-DCs在IL-12分泌和刺激同种异体淋巴细胞反应的能力方面无显著变化,但CD83表达明显较低。hTERT-DCs诱导的特异性CTL对端粒酶阳性靶细胞的细胞毒性高于对阴性靶细胞的细胞毒性。
携带hTERT片段的重组逆转录病毒感染可能损害DCs的成熟,然而,DCs仍保留其激活和刺激淋巴细胞增殖以及启动自体T淋巴细胞产生针对hTERT的特异性CTL的能力。