Wendtner C M, Nolte A, Mangold E, Buhmann R, Maass G, Chiorini J A, Winnacker E L, Emmerich B, Kotin R M, Hallek M
Medizinische Klinik, Klinikum Innenstadt, Germany.
Gene Ther. 1997 Jul;4(7):726-35. doi: 10.1038/sj.gt.3300447.
Gene transfer of the costimulatory molecules B7-1 and B7-2 induces a potent antitumor immune response in a variety of tumor models. B cell neoplasms including multiple myeloma (MM) often show little or no expression of B7 antigens; they are therefore a potential target for this approach. To increase the expression of human B7 genes in MM cells, both genes and the neomycin phosphotransferase gene were packaged into recombinant adeno-associated virus vectors (rAAV). The resulting recombinant viruses rAAV/B7-1, rAAV/B7-2 and rAAV/Neo were used to transduce the MM cell lines LP-1 and RPMI 8226. This allowed the transduction of up to 80% of LP-1 cells 4 days after infection with purified rAAV particles. The response of human allogeneic T cells to rAAV/B7-1 and rAAV/B7-2 transduced, gamma-irradiated LP-1 cells was assessed by [3H]thymidine incorporation, by RT-PCR-based detection of immunostimulatory cytokine transcripts and by ELISA quantification of cytokines in the supernatant. Stimulation of T cells with rAAV/B7-1 or rAAV/B7-2 transduced LP-1 cells resulted in an up to 10-fold increase of T cell proliferation when compared with LP-1 cells transduced with rAAV/Neo. Similar results were obtained with RPMI 8226 cells. Both rAAV/B7-1 and rAAV/B7-2 transduced LP-1 cells stimulated the T cell secretion of IL-2 and IFN-gamma. Furthermore, [51Cr] release assays showed that rAAV/B7-1 or rAAV/B7-2 transduced LP-1 cells induced a cytolytic T cell (CTL) response, in contrast to LP-1 cells transduced with rAAV/Neo. In all assays, the effects of rAAV/B7-1 and rAAV/B7-2 were similar. Taken together, the results show that rAAV-mediated transfer of B7 genes into MM cell lines is able to enhance the antitumor T cell response and to elicit a cytolytic T cell response.
共刺激分子B7-1和B7-2的基因转移在多种肿瘤模型中可诱导强烈的抗肿瘤免疫反应。包括多发性骨髓瘤（MM）在内的B细胞肿瘤通常很少或不表达B7抗原；因此，它们是这种方法的潜在靶点。为了增加MM细胞中人B7基因的表达，将这两个基因和新霉素磷酸转移酶基因包装到重组腺相关病毒载体（rAAV）中。所得的重组病毒rAAV/B7-1、rAAV/B7-2和rAAV/Neo用于转导MM细胞系LP-1和RPMI 8226。在用纯化的rAAV颗粒感染后4天，这使得高达80%的LP-1细胞被转导。通过[3H]胸苷掺入、基于RT-PCR的免疫刺激细胞因子转录本检测以及酶联免疫吸附测定（ELISA）对上清液中细胞因子的定量，评估人同种异体T细胞对rAAV/B7-1和rAAV/B7-2转导的、经γ射线照射的LP-1细胞的反应。与用rAAV/Neo转导的LP-1细胞相比，用rAAV/B7-1或rAAV/B7-2转导的LP-1细胞刺激T细胞导致T细胞增殖增加高达10倍。用RPMI 8226细胞也获得了类似结果。rAAV/B7-1和rAAV/B7-2转导的LP-1细胞均刺激T细胞分泌白细胞介素-2（IL-2）和γ干扰素（IFN-γ）。此外，[51Cr]释放试验表明，与用rAAV/Neo转导的LP-1细胞相比，rAAV/B7-1或rAAV/B7-2转导的LP-1细胞诱导了细胞毒性T细胞（CTL）反应。在所有试验中，rAAV/B7-1和rAAV/B7-2的作用相似。综上所述，结果表明rAAV介导的B7基因向MM细胞系的转移能够增强抗肿瘤T细胞反应并引发细胞毒性T细胞反应。
