Shahabi Vafa, Reyes-Reyes Mariela, Wallecha Anu, Rivera Sandra, Paterson Yvonne, Maciag Paulo
Research and Development, Advaxis, Inc, North Brunswick, NJ, USA.
Cancer Immunol Immunother. 2008 Sep;57(9):1301-13. doi: 10.1007/s00262-008-0463-z. Epub 2008 Feb 14.
Prostate specific antigen (PSA) is a likely immunotherapeutic target antigen for prostate cancer, the second leading cause of cancer-related death in American men. Previously, we demonstrated that attenuated strains of Listeria monocytogenes (Lm) can be used as effective vaccine vectors for delivery of tumor antigens causing regression of established tumors accompanied by strong immune responses toward these antigens in murine models of cancer. In the present study, we have developed and characterized a recombinant live attenuated L. monocytogenes/PSA (Lm-LLO-PSA) vaccine with potential use for the treatment of pCa. Human PSA gene was cloned into and expressed by an attenuated Lm strain. This recombinant bacterial vaccine, Lm-LLO-PSA was tested for stability, virulence, immunogenicity and anti-tumor effects in a murine model for pCa. Immunization with Lm-LLO-PSA was shown to lower the number of tumor infiltrating T regulatory cells and cause complete regression of over 80% of tumors formed by an implanted genetically modified mouse prostate adenocarcinoma cell line, which expressed human PSA. Lm-LLO-PSA was immunogenic in C57BL/6 mice and splenocytes from mice immunized with Lm-LLO-PSA showed significantly higher number of IFN-gamma secreting cells over that of the naïve animals in response to a PSA H2Db-specific peptide, as measured by both, ELISpot and intracellular cytokine staining. In addition, using a CTL assay we show that the T cells specific for PSA were able to recognize and lyse PSA-peptide pulsed target cells in vitro. In a comparison study with two other PSA-based vaccines (a pDNA and a vaccinia vaccine), Lm-LLO-PSA was shown to be more efficacious in regressing established tumors when used in a homologues prime/boost regimen. Together, these results indicate that Lm-LLO-PSA is a potential candidate for pCa immunotherapy and should be further developed.
前列腺特异性抗原(PSA)可能是前列腺癌的一种免疫治疗靶抗原,前列腺癌是美国男性癌症相关死亡的第二大原因。此前,我们证明单核细胞增生李斯特菌(Lm)的减毒株可作为有效的疫苗载体,用于递送肿瘤抗原,在癌症小鼠模型中使已形成的肿瘤消退,并伴随对这些抗原产生强烈的免疫反应。在本研究中,我们开发并鉴定了一种重组减毒活单核细胞增生李斯特菌/PSA(Lm-LLO-PSA)疫苗,其具有用于治疗前列腺癌(pCa)的潜力。将人PSA基因克隆到减毒Lm菌株中并进行表达。在pCa小鼠模型中对这种重组细菌疫苗Lm-LLO-PSA的稳定性、毒力、免疫原性和抗肿瘤作用进行了测试。结果显示,用Lm-LLO-PSA免疫可降低肿瘤浸润性调节性T细胞的数量,并使植入的表达人PSA的转基因小鼠前列腺腺癌细胞系形成的肿瘤中超过80%完全消退。Lm-LLO-PSA在C57BL/6小鼠中具有免疫原性,通过ELISpot和细胞内细胞因子染色检测,用Lm-LLO-PSA免疫的小鼠脾细胞在对PSA H2Db特异性肽作出反应时,分泌γ干扰素的细胞数量显著高于未免疫动物。此外,使用细胞毒性T淋巴细胞(CTL)检测,我们发现针对PSA的T细胞能够在体外识别并裂解PSA肽脉冲处理的靶细胞。在与另外两种基于PSA的疫苗(一种质粒DNA疫苗和一种痘苗疫苗)的比较研究中,当采用同源初免/加强免疫方案时,Lm-LLO-PSA在使已形成的肿瘤消退方面显示出更高的疗效。总之,这些结果表明Lm-LLO-PSA是pCa免疫治疗的一个潜在候选疫苗,应进一步研发。