ELISPOT检测人乳头瘤病毒特异性的记忆T细胞

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(peripheral blood 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方法栓测PBMC中HPV抗原特异性的记忆T细胞.PBMC经表位肽刺激10 d后,细胞数量有明星增加,由最初的4.1 x105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,能够识别抗原连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-cella in persons who spontaneously cleared up the virus (PSCV) after infected the HPV was tested by ELISFOT. Peripheral blood from PSCV (74 months after the clearing up) was collected and isolated the peripheral blood mononuclesr cells (PBMC). PBMC were stimulated with identified epitope-peplide. Tcd days after the cell number was counted. The epilope-pcplide was removed and ihe 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 X 106 from the original number 4.1 x 105. At the eleventh day, tbe cell number increased to4. 65 x 106 which was 11.3-fold before Ibe antigen stimulation. The results of ELISPOT indicated that after ihe 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 cella in PBMC from PSCV infection ws· 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. ELI5-POT could be used to examine HPV-specific memory T cells in peripheral blood.