ELISPOT检测人乳头瘤病毒特异性的记忆T细胞  

Detection of Human Papillomavirus Specific Memory T Cells by ELISPOT

作　　者：王雪莲[1] 卢岩[2] 安春丽[1] 

机构地区：[1]中国医科大学基础医学院病原生物学教研室,辽宁沈阳110001 [2]中国医科大学附属盛京医院院感科,辽宁沈阳110004

出　　处：《微生物学杂志》2011年第4期104-106,共3页Journal of Microbiology

基　　金：国家自然科学基金(81101989);辽宁省教育厅科研项目(L2010579)

摘　　要：应用ELISPOT方法检测人乳头瘤病毒感染后自发清除者外周血中抗原特异性的记忆T细胞。收集人乳头瘤病毒(HPV)感染后自发清除者外周血(病毒清除后74个月),分离外周血单个核细胞(peripheralblood mononuclear cells,PBMC)。体外应用已鉴定的表位肽刺激PBMC,10 d后计数细胞,去除表位肽,继续培养。第11天,ELISPOT方法检测PBMC中HPV抗原特异性的记忆T细胞。PBMC经表位肽刺激10 d后,细胞数量有明显增加,由最初的4.1×105增加为4.2×106。第11天,细胞数量增加为4.65×106,为抗原刺激前细胞数的11.3倍。ELISPOT结果显示,PBMC中的记忆T细胞活化后,能够识别抗原递呈细胞递呈的抗原肽,并分泌IFN-γ。此HPV自发清除者外周血中抗原特异性的记忆T细胞的频数为0.007%。人乳头瘤病毒(HPV)感染后自发清除者外周血存在抗原特异性记忆T细胞,抗原肽可激活记忆T细胞,使之数量增加,分泌IFN-γ。ELISPOT可用于检测外周血中HPV特异性的记忆T细胞。Human papillomavirus（HPV） specific memory T-cells in persons who spontaneously cleared up the virus（PSCV） after infected the HPV was tested by ELISPOT.Peripheral blood from PSCV（74 months after the clearing up） was collected and isolated the peripheral blood mononuclear cells（PBMC）.PBMC were stimulated with identified epitope-peptide.Ten days after the cell number was counted.The epitope-peptide was removed and the cells was continued to be cultured.At the eleventh day,ELISPOT was used to examine HPV specific memory T cells in the PBMC.The results showed the number of PBMC increased 10 days after epitope peptide stimulation to 4.2×106 from the original number 4.1×105.At the eleventh day,the cell number increased to 4.65×106 which was 11.3-fold before the antigen stimulation.The results of ELISPOT indicated that after the activation of memory T cells in PBMC,they could recognize the peptide presented by antigen presenting cells and excrete IFN-γ.The frequency of antigen-specific memory T cells in PBMC from PSCV infection was 0.007%.Therefore,antigen specific memory T cells existed in PBMC in PSCV infection.These memory T cells could be activated and secrete IFN-γ after antigen stimulation.ELISPOT could be used to examine HPV-specific memory T cells in peripheral blood.

关 键 词：ELISPOT 人乳头瘤病毒 记忆T细胞 

分 类 号：R373[医药卫生—病原生物学]