Nilges Katja, Höhn Hanni, Pilch Henryk, Neukirch Claudia, Freitag Kirsten, Talbot P J, Maeurer Markus J
Department of Medical Microbiology, University of Mainz, Germany.
J Virol. 2003 May;77(9):5464-74. doi: 10.1128/jvi.77.9.5464-5474.2003.
Human papillomavirus type 16 (HPV16) E6 and E7 oncoproteins are required for cellular transformation and represent candidate targets for HPV-specific and major histocompatibility complex class I-restricted CD8(+)-T-cell responses in patients with cervical cancer. Recent evidence suggests that cross-reactivity represents the inherent nature of the T-cell repertoire. We identified HLA-A2 binding HPV16 E7 variant peptides from human, bacterial, or viral origin which are able to drive CD8(+)-T-cell responses directed against wild-type HPV16 E7 amino acid 11 to 19/20 (E7(11-19/20)) epitope YMLDLQPET(T) in vitro. CD8(+) T cells reacting to the HLA-A2-presented peptide from HPV16 E7(11-19(20)) recognized also the HLA-A2 binding peptide TMLDIQPED (amino acids 52 to 60) from the human coronavirus OC43 NS2 gene product. Establishment of coronavirus NS2-specific, HLA-A2-restricted CD8(+)-T-cell clones and ex vivo analysis of HPV16 E7 specific T cells obtained by HLA-A2 tetramer-guided sorting from PBL or tumor-infiltrating lymphocytes obtained from patients with cervical cancer showed that cross-reactivity with HPV16 E7(11-19(20)) and coronavirus NS2(52-60) represents a common feature of this antiviral immune response defined by cytokine production. Zero of 10 patients with carcinoma in situ neoplasia and 3 of 18 patients with cervical cancer showed > or =0.1% HPV16 E7-reactive T cells in CD8(+) peripheral blood lymphocytes. In vivo priming with HPV16 was confirmed in patients with cervical cancer or preinvasive HPV16-positive lesions using HLA-A2 tetramer complexes loaded with the E6-derived epitope KLPQLCTEL. In contrast, we could not detect E6-reactive T cells in healthy individuals. These data imply that the measurement of the HPV16 E7(11-19(20)) CD8(+)-T-cell response may reflect cross-reactivity with a common pathogen and that variant peptides may be employed to drive an effective cellular immune response against HPV.
人乳头瘤病毒16型（HPV16）E6和E7癌蛋白是细胞转化所必需的，并且是宫颈癌患者中HPV特异性及主要组织相容性复合体I类限制性CD8⁺ T细胞应答的候选靶点。最近的证据表明，交叉反应性是T细胞库的固有特性。我们从人、细菌或病毒来源中鉴定出了与HLA - A2结合的HPV16 E7变异肽，这些肽在体外能够驱动针对野生型HPV16 E7氨基酸11至19/20（E7(11 - 19/20)）表位YMLDLQPET(T)的CD8⁺ T细胞应答。对来自HPV16 E7(11 - 19(20))且由HLA - A2呈递的肽产生反应的CD8⁺ T细胞，也识别来自人冠状病毒OC43 NS2基因产物的与HLA - A2结合的肽TMLDIQPED（氨基酸52至60）。建立冠状病毒NS2特异性、HLA - A2限制性CD8⁺ T细胞克隆，并对通过HLA - A2四聚体引导分选从宫颈癌患者的外周血单个核细胞（PBL）或肿瘤浸润淋巴细胞中获得的HPV16 E7特异性T细胞进行体外分析，结果表明与HPV16 E7(11 - 19(20))和冠状病毒NS2(52 - 60)的交叉反应性是这种由细胞因子产生所定义的抗病毒免疫应答的一个共同特征。10例原位癌患者中无1例、18例宫颈癌患者中有3例在CD8⁺外周血淋巴细胞中显示出≥0.1%的HPV16 E7反应性T细胞。使用负载有E6衍生表位KLPQLCTEL的HLA - A2四聚体复合物，在宫颈癌或HPV16阳性的癌前病变患者中证实了HPV16的体内启动。相比之下，我们在健康个体中未检测到E6反应性T细胞。这些数据表明，对HPV16 E7(11 - 19(20)) CD8⁺ T细胞应答的检测可能反映了与一种常见病原体的交叉反应性，并且变异肽可用于驱动针对HPV的有效细胞免疫应答。
