Tetramer技术检测人乳头瘤病毒抗原特异性T细胞条件的优化

人乳头瘤病毒(human papillomavirus,HPV)感染是宫颈癌发生的首要因素。针对病毒早期蛋白E6的特异性细胞毒性T淋巴细胞(cytotoxic Tlymphocyte,CTL)在清除HPV感染细胞和病毒转化形成的肿瘤细胞过程中发挥重要作用,因此检测体内抗原特异性CTL的频数和功能有助于了解病毒感染者或宫颈癌患者体内的特异性细胞免疫反应。利用加载HPV16 E6表位抗原肽(E6 133-142;HNIRGRWTGR)的HLA-A6801四聚体(Tetramer),即HPV16 E6 133-142/HLA-A6801-PE四聚体,通过检测混有HPV特异性CTL的PBMC标本,优化Tetramer染色的实验条件,探讨染色的最佳温度及Tetramer浓度。结果显示,染色温度(4℃、室温及37℃)对Tetramer与CTL的结合无明显影响。Tetramer稀释度为1∶1 600时,HPV特异性CTL荧光强度保持高水平,且非特异性染色少,为最佳染色浓度。研究结果为进一步检测HPV感染者或宫颈癌患者体内抗原特异性的CTL打下基础。

Optimization of Tetramer Staining Condition to Detect of Human Papillomavirus Specific T Cells

The infection of human papillomavirus(HPV) is the primary pathogenic factor in the process of occurrence of cervical cancer.The specific cytotoxic T lymphocyte(CTL) to HPV early protein E6 is important in the clearance of the virus-infected cells or the virus-transformed cells.Detecting the frequency and the function of HPV-specific CTL is conducive to learn the cellular immunity in virus-infected women or cervical cancer patients.The optimizing staining temperature and titration of tetramer by detecting PB...