GENERATION OF TUMOR INFILTRATING LYMPHOCYTES (TIL) FROM HHV-6 DNA INDUCED TUMORS

我们最近发现了HHV-6基因组和一个亚基因组克隆， PZVHI4，可以从形态上转化NIH 3T3细胞，使其能够产生 裸鼠体内的肿瘤。在这个项目中，我们还观察到这些 成瘤细胞可在免疫活性小鼠体内产生肿瘤 转移到肺部。由HHV-6全基因组和 亚基因组克隆pZVHI4命名为G-2TS和L4-2TS， 分别进行了分析。我们已经从这些肿瘤中产生了TIL，并将它们扩大到 用重组IL-2培养。直到把不同的目标分解到 (51)铬释放试验。G-2TS肿瘤来源的TIL特异性地裂解其 自体靶点，并在6个月期间保持其特异性。 然而，在培养的早期阶段，TIL来源于L4-2TS肿瘤 裂解的自体肿瘤和包括Yac-1在内的同种异体靶点 淋巴瘤。在培养几天后，这些L4-2TS TIL失去了裂解能力 自体目标的能力，但甚至继续溶解YAC-I目标 经过56天的培养。这些TIL的表型为Thy 1.2，Lyt-2 和L3T4-。这些研究表明，HHV-6DNA转染的NIH 3T3细胞 获得致瘤和转移潜能以及荷瘤宿主 可以对这类肿瘤产生免疫反应。

We have recently shown that the HHV-6 genome and a subgenomic clone, pZVHI4, can morphologically transform NIH 3T3 cells which can produce tumors in nude mice. In this project, we also observed that these tumorigenic cells could produce tumors in immunocompetent mice and metastasize to the lung. The tumors induced by the entire HHV-6 genome and the subgenomic clone, pZVHI4, were designated as G-2TS and l4-2TS, respectively. We have generated TIL from these tumors and expanded them in culture with recombinant IL-2. These TIL lysed various targets in (51)Cr-release assays. TIL derived from G-2TS tumors lysed specifically its autologous targets and maintained their specificity over 6-month period. However, at the early phase of culture, TIL derived from l4-2TS tumors lysed autologous tumors as well as allogeneic targets including Yac-1 lymphoma. After several days in culture, these l4-2TS TIL lost their lytic capability of autologous targets, but continued to lyse Yac-I target even after 56 days of culture. Phenotypically, these TIL were Thy 1.2+, Lyt-2+ and L3T4-. These studies indicate that HHV-6 DNA transfected NIH 3T3 cells acquire both tumorigenic and metastatic potential and tumor bearing hosts can mount immune responses against such tumors.