HPV疫苗接种者与宫颈癌前病变及宫颈癌患者的HPV16特异性免疫特征

HPV16-specific immune characteristics in HPV vaccinees,patients with cervical precancerous lesions or cervical cancer

目的比较性研究HPV疫苗接种者与宫颈癌前病变及宫颈癌患者针对HPV16病毒产生的特异性免疫特征。方法选取6名女性HPV疫苗接种者、5名宫颈癌前病变或宫颈癌患者及8名健康女性志愿者为研究对象,采集研究对象的抗凝血并分离外周血单个核细胞(PBMCs)和血浆,用HPV16 L1蛋白多肽库作为刺激物对其进行体外培养9-10天后利用流式细胞术检测样本分泌细胞因子IFN-γ、IL-2及TNF-α的CD4^(+)及CD8^(+)T淋巴细胞百分比,同时检测研究对象血浆中的HPV16 IgG抗体水平,分析比较不同组别人群针对HPV16的特异性免疫特征。结果不同组别人群血浆中的IgG水平存在差异,疫苗接种者的IgG水平高于健康人和宫颈癌前病变及宫颈癌患者,差异具有统计学意义(p<0.05);疫苗接种者针对HPV16的特异性CD4^(+)T细胞免疫水平较健康人相比明显升高,差异具有统计学意义(p<0.05);整体人群体内针对HPV16的特异性T细胞免疫水平与其抗体水平之间存在正相关性(Pearson检验,r>0.4,p<0.05)。结论不同组别人群针对HPV的特异性免疫反应具有明显的特征差异,HPV疫苗接种者体内存在较高的特异性抗体及CD4^(+)T细胞免疫。

Objective To comparative investigate the HPV16 virus-specific immune characteristics of HPV vaccinees,cervical precancerous lesions patients and cervical cancer patients.Methods Six female HPV vaccinees,five patients with cervical precancerous lesions or cervical cancer and 8 healthy female volunteers from Beijing were recruited as subjects.Peripheral anticoagulated blood specimens were collected from these subjects and the peripheral blood mononuclear cells(PBMCs)and plasma were isolated.After culturing in vitro with the stimulation of L1 protein peptide pool derived from HPV16 for 9-10 days,the percentage of CD4^(+) and CD8^(+)-T lymphocytes secreting cytokines IFN-γ,IL-2 and TNF-αin the PBMCs were detected by flow cytometry.The level of HPV16 IgG antibody in plasma was determined by ELISA.Results The IgG levels of vaccinees were significantly higher than those of healthy donors as well as patients with cervical precancerous lesions or cervical cancer.Moreover,the specific CD4^(+)T cell immune level of vaccinees against HPV16 was significantly higher than that of healthy controls.Furthermore,there was a positive correlation between the level of specific T cell immunity and the level of antibody against HPV16 in all the donors(Pearson,r>0.4,p<0.05).Conclusions The virus-specific immune responses,i.e.humoral responses as well as cellular immune responses are presenting diversely among different HPV-related subjects.HPV vaccinees possess robust humoral immunity and CD4^(+)T cell responses.