Raykov Zahari, Savelyeva Larissa, Balboni Ginette, Giese Thomas, Rommelaere Jean, Giese Nathalia A
Department of Surgery, Medical School, University of Heidelberg, INF 116, 60120 Heidelberg, Germany.
J Virol. 2005 Mar;79(6):3517-24. doi: 10.1128/JVI.79.6.3517-3524.2005.
Due to their oncolytic properties and apathogenicity, autonomous parvoviruses have attracted significant interest as possible anticancer agents. Recent preclinical studies provided evidence of the therapeutic potential of minute virus of mice prototype strain (MVMp) and its recombinant derivatives. In a murine model of hemangiosarcoma, positive therapeutic outcome correlated with high intratumoral expression of MVMp-encoded genes in tumors and lymphoid organs, especially in tumor-draining lymph nodes. The source and relevance of this extratumoral expression, which came as a surprise because of the known fibrotropism of MVMp, remained unclear. In the present study, we investigated (i) whether the observed expression pattern occurs in different tumor models, (ii) which cell population is targeted by the virus, and (iii) the immunological consequences of this infection. Significant MVMp gene expression was detected in lymphoid tissues from infected tumor-free as well as melanoma-, lymphoma-, and hemangiosarcoma-bearing mice. This expression was especially marked in lymph nodes draining virus-injected tumors. Fluorescent in situ hybridization analysis, multicolor fluorescence-activated cell sorting, and quantitative reverse transcription-PCR revealed that MVMp was expressed in rare subpopulations of CD11b (Mac1)-positive cells displaying CD11c+ (myeloid dendritic cells [MDC]) or CD45B (B220+ [B1 lymphocytes]) markers. Apart from the late deletion of cytotoxic memory cells (CD8+ CD44+ CD62L-), this infection did not lead to significant alteration of the immunological profile of cells populating lymphoid organs. However, subtle changes were detected in the production of specific proinflammatory cytokines in lymph nodes from virus-treated animals. Considering the role of B1 lymphocytes and MDC in cancer and immunological surveillance, the specific ability of these cell types to sustain parvovirus-driven gene expression may be exploited in gene therapy protocols.
由于其溶瘤特性和无致病性,自主细小病毒作为潜在的抗癌药物引起了广泛关注。最近的临床前研究提供了小鼠微小病毒原型株(MVMp)及其重组衍生物具有治疗潜力的证据。在血管肉瘤的小鼠模型中,积极的治疗结果与肿瘤和淋巴器官中MVMp编码基因的高肿瘤内表达相关,特别是在引流肿瘤的淋巴结中。这种肿瘤外表达的来源和相关性尚不清楚,因为已知MVMp具有嗜纤维性,这一现象令人惊讶。在本研究中,我们调查了:(i)在不同肿瘤模型中是否会出现观察到的表达模式;(ii)病毒靶向的细胞群体是什么;(iii)这种感染的免疫后果。在未感染肿瘤以及患有黑色素瘤、淋巴瘤和血管肉瘤的小鼠的淋巴组织中检测到了显著的MVMp基因表达。这种表达在引流病毒注射肿瘤的淋巴结中尤为明显。荧光原位杂交分析、多色荧光激活细胞分选和定量逆转录PCR显示,MVMp在显示CD11c+(髓样树突状细胞 [MDC])或CD45B(B220+ [B1淋巴细胞])标记的CD11b(Mac1)阳性细胞的罕见亚群中表达。除了细胞毒性记忆细胞(CD8+ CD44+ CD62L-)的晚期缺失外,这种感染并未导致淋巴器官中细胞免疫谱的显著改变。然而,在病毒处理动物的淋巴结中,特定促炎细胞因子的产生检测到了细微变化。考虑到B1淋巴细胞和MDC在癌症和免疫监视中的作用,这些细胞类型维持细小病毒驱动基因表达的特定能力可在基因治疗方案中加以利用。