宫颈癌患者外周血HPV16抗原特异性CD8~+细胞毒性T细胞的检测及意义

目的:初步探讨人乳头瘤病毒16型(HPV16)阳性宫颈癌患者细胞免疫状况,了解其外周血抗原特异性CD8~+细胞毒性T细胞的水平.方法:采取免疫荧光标染重组MHC Ⅰ类分子-肽五聚体技术,运用流式细胞术定量检测患者和正常人外周血CD8~+细胞毒性T细胞、记忆性细胞毒性T细胞、活化细胞毒性T细胞及HPV16抗原特异性CD8~+细胞毒性T细胞.结果:宫颈癌患者CD8~+细胞毒性T细胞计数、记忆性细胞毒性T细胞计数、体内抗原特异性CD8~+细胞毒性T细胞计数和对照组比较差异无统计学意义,但活化细胞毒性T细胞计数低于对照组(P=0.322),体外经抗原肽刺激后抗原特异性CD8~+细胞毒性T细胞计数高于对照组(P=0.0068).结论:HPV16 阳性宫颈癌患者外周血中活化细胞毒性T细胞数目减少,表明患者细胞免疫功能受到一定的影响,通过抗原表位有效的诱导是提高患者的特异性细胞免疫功能的重要途径,运用五聚体技术检测宫颈癌患者抗原特异性细胞毒性T细胞,可初步反映患者特异性细胞免疫状态并有利于进行复发和预后分析.

Significance of Quantification of Peripheral Blood HPV16 Antigen-specific CD8~+ Cytotoxic T Cells in Patients with Cervical Cancer

Objeotive:The cellular immune response and the antigen-specific CD8~+ cytotoxic T cells (CTLs) in peripheral blood of patients with cervical cancer associated with infection of Human papilloma virus type 16 were investigated. Methods:CD8~+ CTLs, CD8~+ CD45 RO~+ CTLs, and CD8~+ CD69~+ CTLs ( activated CTLs) were detected by flow cytometry, and antigen-specific CD8~+ CTLs were stained with fluorescent-la-beled recombinant MHC class I -peptide pentamer complex. Results;There was no significant difference in the percentage of CD8~+CTLs, CD8~+ CD45RO~+ CTLs, and antigen-specific CD8~+ CTLs between peripheral blood of the patients and that of the control group, however, the percentage of CD8~+ CD69~+ CTLs was lower than that of control group ( P = 0.332 ). Antigen-specific CD8~+ CTLs post-stimulus by peptide in vitro was higher than that of control group ( P = 0.0068 ). Conclusions:The decrease of activated CTLs in peripheral blood of patients with cervical cancer associated with infection by Human papilloma virus type 16 reflects the hampered callular immune function of patients. It is an important pathway to induce antigen-specific CTLs with epitopes for improvement of specific cellular immunity. Detection of antigen-specific CTLs by MHC class I -peptide pentamer complex can probably reflect the specific cellular immunity of patients, and be used for prediction of recurrence and prognosis.