Rutebemberwa Alleluiah, Currier Jeffrey R, Jagodzinski Linda, McCutchan Francine, Birx Deborah, Marovich Mary, Cox Josephine H
Henry M. Jackson Foundation and the US Military HIV Research Program, Rockville, Maryland 20850, USA.
AIDS. 2005 Jul 22;19(11):1165-72. doi: 10.1097/01.aids.0000176216.02743.98.
To compare the ability of three Env (15-mer) peptide sets derived from the HIV-1 MN, the subtype B consensus, and the group M consensus to detect HIV-1 specific interferon (IFN)-gamma responses in HIV-1 subtype B infected subjects.
Peripheral blood mononuclear cells were obtained from 17 HIV-1 subtype B seropositive and 5 HIV-1 seronegative subjects. Peptide matrices comprising each peptide set were used in IFN-gamma Elispot assays to screen for T cell epitopes. Following matrix deconvolution, individual peptides were analyzed by IFN-gamma intracellular cytokine-staining to confirm and characterize the responding cells.
HIV specific IFN-gamma responses were detected in 17 of 17 HIV-1 seropositive and none of 5 HIV-1 seronegative subjects by Elispot. Within the 17 HIV-1 seropositives, 16, 14, and 11 subjects responded to MN, B consensus, and group M env peptides, respectively. Responses were confirmed by intracellular cytokine analysis in 14 subjects and were in the CD3CD8 compartment. Cross-recognition of 'equivalent' peptides (i.e., peptides mapping to the same sequence region from the three peptide sets) was observed in 9 of 17 subjects. Peptide set specific responses to individual peptides were also observed; 11, 1, and 1 subjects demonstrated peptide set specific responses to MN, B consensus, and consensus group M, respectively.
MN derived Env peptides were better able to detect HIV-1 specific CD8 T cell responses, many of which were not detectable by the equivalent clade or group consensus peptides. No single peptide set detected all the IFN-gamma responses within an individual. These results demonstrate the importance of reagent selection for monitoring of HIV responses in HIV-1 infected individuals and subsequently vaccine recipients.
比较源自HIV-1 MN、B亚型共识序列和M组共识序列的三种Env(15肽)肽组在检测HIV-1 B亚型感染受试者中HIV-1特异性干扰素(IFN)-γ应答的能力。
从17名HIV-1 B亚型血清阳性和5名HIV-1血清阴性受试者中获取外周血单个核细胞。在IFN-γ酶联免疫斑点试验中使用包含每个肽组的肽矩阵来筛选T细胞表位。在矩阵去卷积后,通过IFN-γ细胞内细胞因子染色分析单个肽以确认并表征应答细胞。
通过酶联免疫斑点试验在17名HIV-1血清阳性受试者中的17名检测到HIV特异性IFN-γ应答,而5名HIV-1血清阴性受试者中均未检测到。在17名HIV-1血清阳性者中,分别有16、14和11名受试者对MN、B亚型共识序列和M组Env肽有应答。在14名受试者中通过细胞内细胞因子分析确认了应答,且应答位于CD3CD8区室。在17名受试者中的9名观察到“等效”肽(即来自三个肽组映射到相同序列区域的肽)的交叉识别。也观察到对单个肽的肽组特异性应答;分别有11、1和1名受试者对MN肽组、B亚型共识序列肽组和M组共识序列肽组表现出肽组特异性应答。
源自MN的Env肽更能检测HIV-1特异性CD8 T细胞应答,其中许多应答无法通过等效的进化枝或组共识肽检测到。没有单个肽组能检测到个体内所有的IFN-γ应答。这些结果证明在监测HIV-1感染个体以及随后的疫苗接种者中的HIV应答时试剂选择非常重要。