Suzuki T, Piche A, Kasono K, Xiang J, Gomez-Navarro J, Moriuchi S, Krisky D M, Oligino T, Glorioso J C, Curiel T J, Curiel D T
Gene Therapy Program, University of Alabama at Birmingham, Alabama, 35294-3000, USA.
Biochem Biophys Res Commun. 1998 Nov 27;252(3):686-90. doi: 10.1006/bbrc.1998.9685.
Subgroups of the B cell malignancies are known to be associated with Epstein-Barr virus (EBV) infection, especially in immunocompromised patients. These are fatal and refractory to conventional antineoplastic therapy. B cells are usually post-mitotic cells and even mitogen activated or transformed B cells have shown relative resistance against viral mediated gene transfer. To address this issue, we employed a replication-defective herpes simplex virus-1 (HSV-1) to mediate gene transfer into EBV-transformed B cells. The virus expresses the herpes simplex virus thymidine kinase (HSV-TK) and the E. coli lacZ reporter genes and is designated T0Z.1. We used the lymphoblastoid cell line SWEIG as a model for human EBV-related B cell malignancy. This cell line was established by in vitro EBV infection of primary human peripheral blood mononuclear cells. When SWEIG cells were infected with T0Z.1, X-gal staining revealed lacZ expression in more than 20% cells even at multiplicity of infection (MOI) as low as 1 and the expression persisted for at least one week. Ganciclovir (GCV) administration after T0Z.1 infection effectively decreased the number of the infected tumor cells in a dose-responsive manner. Viral toxicity was analyzed by cell proliferation assay (MTS assay) and found to be little even at 10 MOI infection. Three MOI of the virus yielded maximum antineoplastic effect and more than 50% tumor cells were killed by HSV-TK/GCV. These results suggest the potential utility of replication-defective HSV-1 for the treatment of EBV-related B cell malignancies.
已知B细胞恶性肿瘤的亚组与爱泼斯坦-巴尔病毒（EBV）感染有关，尤其是在免疫功能低下的患者中。这些肿瘤是致命的，并且对传统的抗肿瘤治疗具有抗性。B细胞通常是有丝分裂后细胞，即使是有丝分裂原激活或转化的B细胞对病毒介导的基因转移也表现出相对抗性。为了解决这个问题，我们采用了复制缺陷型单纯疱疹病毒1型（HSV-1）介导基因转移到EBV转化的B细胞中。该病毒表达单纯疱疹病毒胸苷激酶（HSV-TK）和大肠杆菌lacZ报告基因，命名为T0Z.1。我们使用淋巴母细胞系SWEIG作为人类EBV相关B细胞恶性肿瘤的模型。该细胞系通过体外EBV感染原代人外周血单核细胞建立。当SWEIG细胞用T0Z.1感染时，X-gal染色显示即使在感染复数（MOI）低至1时，超过20%的细胞中有lacZ表达，并且该表达持续至少一周。T0Z.1感染后给予更昔洛韦（GCV）以剂量反应方式有效减少了感染肿瘤细胞的数量。通过细胞增殖测定（MTS测定）分析病毒毒性，发现即使在10 MOI感染时毒性也很小。病毒的三个MOI产生最大抗肿瘤效果，超过50%的肿瘤细胞被HSV-TK/GCV杀死。这些结果表明复制缺陷型HSV-1在治疗EBV相关B细胞恶性肿瘤方面具有潜在效用。
