Xue Song, Sun Ying-hao, Gao Jian-ping, Xu Xiao-feng, Zhang Zheng-yu, Xu Chuan-liang, Zhu Xue-jun, Fan Zhen-fang
Department of Urology, Nanjing Clinical School, The Second Military Medical University/Nanjing General Hospital of Nanjing Military Region, Nanjing, Jiangsu 210002, China.
Zhonghua Nan Ke Xue. 2010 May;16(5):410-4.
To study the effectiveness of freeze-thaw antigens and acid eluted peptide antigens extracted from tumor cell-pulsed dendritic cells (DC) in inducing prostate cancer-specific cytotoxic T lymphocytes (CTL) in vitro.
Tumor antigens were extracted from the prostate cancer cell line PC-3 with the repeated freeze-thaw and weak acid elution methods. Peripheral blood mononuclear cells were cultured with recombinant human GM-CSF and IL-4 for inducing DCs in vitro. Then the DCs were pulsed with the two kinds of prostate cancer tumor antigens respectively and cultured with T cells for inducing CTLs. The activity of the tumor-specific CTLs were detected by LDH release assay.
The protein content in the tumor antigens obtained from PC-3 (2 x 10(7)) by citric acid-phosphate buffer elution and that by the repeated freeze-thaw method were (212.2 +/- 7.9) microg and (963.0 +/- 25.3) microg, respectively. The two kinds of prostate cancer antigens-pulsed DCs had a significant role in inducing the PC-3 cell-specific CTLs, and the CTLs induced by acid-eluted peptide antigen-pulsed DCs exhibited an even more significant tumor-specific cytotoxicity than those induced by repeated freeze-thaw ([60.4 +/- 5.52]% vs. [43.7 +/- 4.11]%, P < 0.01).
Both the weak acid elution and repeated freeze-thaw methods for extracting prostate cancer antigens can be used for in vitro sensitization of DCs. The DCs pulsed by either of the two kinds of antigens can activate CTLs, and the antigens extracted by weak acid elution are even more effective.
研究从肿瘤细胞脉冲树突状细胞(DC)中提取的冻融抗原和酸洗脱肽抗原在体外诱导前列腺癌特异性细胞毒性T淋巴细胞(CTL)的有效性。
采用反复冻融法和弱酸洗脱法从前列腺癌细胞系PC-3中提取肿瘤抗原。外周血单个核细胞用重组人GM-CSF和IL-4体外培养诱导DC。然后分别用两种前列腺癌肿瘤抗原脉冲DC,并与T细胞共培养诱导CTL。通过乳酸脱氢酶释放试验检测肿瘤特异性CTL的活性。
用柠檬酸-磷酸盐缓冲液洗脱法和反复冻融法从PC-3(2×10⁷)中获得的肿瘤抗原中的蛋白质含量分别为(212.2±7.9)μg和(963.0±25.3)μg。两种前列腺癌抗原脉冲DC在诱导PC-3细胞特异性CTL方面具有显著作用,酸洗脱肽抗原脉冲DC诱导的CTL比反复冻融诱导的CTL表现出更显著的肿瘤特异性细胞毒性([60.4±5.52]%对[43.7±4.11]%,P<0.01)。
弱酸洗脱法和反复冻融法提取前列腺癌抗原均可用于DC的体外致敏。两种抗原脉冲的DC均可激活CTL,且弱酸洗脱提取的抗原效果更佳。