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

我们最近发现HHV-6基因组和亚基因组克隆， pZVHI 4可使NIH 3 T3细胞发生形态学转化， 裸鼠肿瘤。在这个项目中，我们还观察到， 致瘤细胞可以在免疫活性小鼠中产生肿瘤， 转移到肺部由整个HHV-6基因组诱导的肿瘤， 将亚基因组克隆pZVHI 4命名为G-2 TS和14 - 2 TS， 分别我们已经从这些肿瘤中产生了TIL，并将其扩增， 用重组IL-2培养。这些TIL裂解各种靶点， （51）Cr释放测定。来源于G-2 TS肿瘤的TIL特异性地裂解其细胞因子， 自体靶点，并在6个月内保持其特异性。 然而，在培养的早期阶段，来自l4- 2 TS肿瘤的TIL 裂解的自体肿瘤以及包括Yac-1在内的同种异体靶标 淋巴瘤在培养几天后，这些14 - 2 TS TIL失去了它们的溶解性。 能力的自体目标，但继续裂解Yac-I目标，甚至 培养56天后。表型上，这些TIL为Thy 1.2+，Lyt-2+， 和L3 T4-。这些研究表明，HHV-6 DNA转染NIH 3 T3细胞 获得致瘤性和转移性潜能以及荷瘤宿主 可以产生针对这种肿瘤的免疫反应。

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.