Scholten Kirsten B J, Schreurs Marco W J, Ruizendaal Janneke J, Kueter Esther W M, Kramer Duco, Veenbergen Sharon, Meijer Chris J L M, Hooijberg Erik
Department of Pathology, VU University Medical Center, de Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
Clin Immunol. 2005 Feb;114(2):119-29. doi: 10.1016/j.clim.2004.11.005.
Human papilloma virus (HPV) type 16 infections of the genital tract are associated with the development of cervical cancer (CxCa) in women. HPV16-derived oncoproteins E6 and E7 are expressed constitutively in these lesions and might therefore be attractive candidates for T-cell-mediated adoptive immunotherapy. However, the low precursor frequency of HPV16E7-specific T cells in patients and healthy donors hampers routine isolation of these cells for adoptive transfer. To overcome this problem, we have isolated T cell receptor (TCR) genes from four different HPV16E7-specific healthy donor and patient-derived human cytotoxic T lymphocyte (CTL) clones. We examined whether genetic engineering of peripheral blood-derived CD8+ T cells in order to express HPV16E711-20-specific TCRs is feasible for adoptive transfer purposes. Reporter cells (Jurkat/MA) carrying a transgenic TCR were shown to bind relevant but not irrelevant tetramers. Moreover, these TCR-transgenic Jurkat/MA cells showed reactivity towards relevant target cells, indicating proper functional activity of the TCRs isolated from already available T cell clones. We next introduced an HPV16E711-20-specific TCR into blood-derived, CD8+ recipient T cells. Transgenic CTL clones stained positive for tetramers presenting the relevant HPV16E711-20 epitope and biological activity of the TCR in transduced CTL was confirmed by lytic activity and by interferon (IFN)-gamma secretion upon antigen-specific stimulation. Importantly, we show recognition of the endogenously processed and HLA-A2 presented HPV16E711-20 CTL epitope by A9-TCR-transgenic T cells. Collectively, our data indicate that HPV16E7 TCR gene transfer is feasible as an alternative strategy to generate human HPV16E7-specific T cells for the treatment of patients suffering from cervical cancer and other HPV16-induced malignancies.
生殖道16型人乳头瘤病毒（HPV）感染与女性宫颈癌（CxCa）的发生相关。HPV16衍生的癌蛋白E6和E7在这些病变中持续表达，因此可能是T细胞介导的过继性免疫治疗的有吸引力的候选对象。然而，患者和健康供体中HPV16E7特异性T细胞的前体频率较低，阻碍了这些细胞用于过继性转移的常规分离。为克服这一问题，我们从4个不同的HPV16E7特异性健康供体和患者来源的人细胞毒性T淋巴细胞（CTL）克隆中分离了T细胞受体（TCR）基因。我们研究了对外周血来源的CD8+ T细胞进行基因工程改造以表达HPV16E711-20特异性TCR用于过继性转移目的是否可行。携带转基因TCR的报告细胞（Jurkat/MA）显示能结合相关而非无关的四聚体。此外，这些TCR转基因Jurkat/MA细胞对相关靶细胞有反应性，表明从已有T细胞克隆中分离的TCR具有适当的功能活性。接下来，我们将HPV16E711-20特异性TCR导入血液来源的CD8+受体T细胞。转基因CTL克隆对呈现相关HPV16E711-20表位的四聚体染色呈阳性，转导的CTL中TCR的生物学活性通过裂解活性以及抗原特异性刺激后的干扰素（IFN）-γ分泌得以证实。重要的是，我们展示了A9-TCR转基因T细胞对内源性加工并由HLA-A2呈递的HPV16E711-20 CTL表位的识别。总体而言，我们的数据表明HPV16E7 TCR基因转移作为一种替代策略是可行的，可用于产生人HPV16E7特异性T细胞，以治疗宫颈癌和其他HPV16诱导的恶性肿瘤患者。
