Greiner S, Humrich J Y, Thuman P, Sauter B, Schuler G, Jenne L
Department of Dermatology, University Hospital Erlangen, Erlangen, Germany.
Clin Exp Immunol. 2006 Nov;146(2):344-53. doi: 10.1111/j.1365-2249.2006.03177.x.
Vaccinia virus (VV) has been tested as oncolytic virus against malignant melanoma in clinical trials for more than 40 years. Until now, mainly strains comparable to viral strains used for smallpox vaccination have been probed for anti-tumoral therapy. We have shown recently that the wild-type strain Western Reserve (WR) can interfere with crucial functions of monocyte-derived dendritic cells (DCs). Our aim was to examine whether viral immune evasion mechanisms might be responsible for the ineffectiveness of WR-based vaccination strategies and whether the highly attenuated strain modified virus Ankara (MVA) differs from WR with respect to its possible immunostimulatory capacity after intratumoral injection. Using in vitro experiments, we compared the effect of both strains on melanoma cells and on local bystander DCs. We found that both VV-strains infected melanoma cells efficiently and caused disintegration of the actin cytoskeleton, as shown by fluorescence microscopy. In addition, both VV-strains caused apoptotic cell death in melanoma cells after infection. In contrast to MVA, WR underwent a complete viral replication cycle in melanoma cells. Bystander DCs were consecutively infected by newly generated WR virions and lost their capacity to induce allogeneic T cell proliferation. DCs in contact with MVA-infected melanoma cells retained their capacity to induce T cell proliferation. Immature DCs were capable of phagocytosing MVA-infected melanoma cells. Priming of autologous CD8(+) T cells by DCs that had phagocytosed MVA-infected, MelanA positive melanoma cells resulted in the induction of T cell clones specifically reactive against the model antigen MelanA as shown by enzyme-linked immunospot (ELISPOT) analysis. We conclude that the clinical trials with oncolytic wild-type VV failed probably because of suppression of bystander DCs and consecutive suppression of T cell-mediated anti-melanoma immunity. The attenuated VV-strain MVA facilitates the generation of tumour associated antigen (TAA)-specific T cell response as it is oncolytic for melanoma cells, but non-toxic for DC, and should be a promising candidate for intralesional metastatic melanoma therapy.
牛痘病毒(VV)作为溶瘤病毒用于恶性黑色素瘤的临床试验已超过40年。到目前为止,主要是对与用于天花疫苗接种的病毒株相当的毒株进行抗肿瘤治疗研究。我们最近发现,野生型毒株西储株(WR)可干扰单核细胞衍生树突状细胞(DC)的关键功能。我们的目的是研究病毒免疫逃逸机制是否可能是基于WR的疫苗接种策略无效的原因,以及高度减毒的毒株改良安卡拉病毒(MVA)在瘤内注射后其可能的免疫刺激能力是否与WR不同。通过体外实验,我们比较了这两种毒株对黑色素瘤细胞和局部旁观者DC的影响。我们发现,两种VV毒株均能有效感染黑色素瘤细胞,并导致肌动蛋白细胞骨架解体,荧光显微镜检查显示了这一点。此外,两种VV毒株在感染后均导致黑色素瘤细胞发生凋亡性细胞死亡。与MVA不同,WR在黑色素瘤细胞中经历了完整的病毒复制周期。旁观者DC被新产生的WR病毒粒子连续感染,并失去了诱导同种异体T细胞增殖的能力。与感染MVA的黑色素瘤细胞接触的DC保留了其诱导T细胞增殖的能力。未成熟DC能够吞噬感染MVA的黑色素瘤细胞。吞噬感染MVA的、MelanA阳性黑色素瘤细胞的DC对自体CD8(+) T细胞的致敏导致诱导出对模型抗原MelanA具有特异性反应的T细胞克隆,酶联免疫斑点(ELISPOT)分析显示了这一点。我们得出结论,溶瘤野生型VV的临床试验失败可能是因为旁观者DC受到抑制以及随后T细胞介导的抗黑色素瘤免疫受到抑制。减毒VV毒株MVA促进肿瘤相关抗原(TAA)特异性T细胞反应的产生,因为它对黑色素瘤细胞具有溶瘤作用,但对DC无毒,应该是瘤内转移性黑色素瘤治疗的一个有前景的候选药物。