Vaccine and Drug Combination Therapy for Human Cancers

某些化疗方案触发癌细胞死亡，同时诱导树突细胞成熟和随后的免疫应答。然而，化疗诱导的免疫原性细胞死亡（ICD）迄今为止仅限于选择药剂。相反，几种化疗药物调节抗肿瘤免疫应答，尽管不诱导经典ICD。此外，在许多情况下，肿瘤细胞在治疗后不会死亡。在这里，使用多西他赛，最广泛使用的癌症化疗药物之一，作为一个模型，我们检查了肿瘤细胞的表型和功能的后果，不死于免疫原性细胞死亡。多西他赛治疗肿瘤细胞不诱导ATP或HMGB 1分泌或细胞死亡。然而，钙网蛋白暴露在化疗治疗后检查的所有细胞系中观察到。多西他赛治疗后，CEA、MUC-1或PSA特异性CD 8 + CTL的杀伤作用显著增强。这种杀伤与抗原加工机制的组分增加有关，并且主要由钙网蛋白膜易位介导，如钙网蛋白、PERK或钙网蛋白阻断肽的功能性敲低所确定。通过连续暴露于多西他赛来选择多西他赛耐药细胞系（MDR-1+，CD 133+）。这些细胞，而耐多西他赛的直接细胞生长抑制作用，不耐的化学调节作用，导致增强的CTL杀伤。我们提供了“免疫原性调节”的操作定义，其中肿瘤细胞暴露于非致死/亚致死剂量的化疗改变肿瘤表型，使肿瘤对CTL杀伤更敏感。这些观察结果与免疫原性细胞死亡是不同的和互补的，并突出了化疗可以与免疫治疗联合使用的机制。铂双联化疗，包括顺铂加长春瑞滨，是非小细胞肺癌（NSCLC）的标准治疗。已证明亚致死暴露于某些化疗剂可改变人肿瘤细胞的表型或生物学，使其对细胞毒性T淋巴细胞（CTL）介导的裂解更敏感。然而，顺铂/长春瑞滨对肿瘤对T细胞毒性的敏感性的影响及其分子机制尚未完全阐明。在这里，我们研究了这种化疗对生长的影响，细胞表面表型，和CTL介导的裂解五种不同的人肺癌细胞系在体外，并研究了与增强CTL敏感性的分子机制。这些研究表明，人肺肿瘤细胞亚致死暴露于铂双联体调节肿瘤细胞表型，并增加敏感性MHC限制性穿孔素/颗粒酶介导的CTL杀伤。这些研究还表明，暴露于化疗显着降低TGF-β/IL-8的蛋白质分泌比。我们检查了两种肺肿瘤细胞系的基因表达谱，以确定响应亚致死顺铂/长春瑞滨的共享基因签名，并发现仅16种转录物的协调表达，包括用于细胞因子/趋化因子表达和凋亡的转录物，如TNF-α、IL 8、CXCL 5和BCL-2样基因。总之，这些结果表明，亚致死暴露于顺铂/长春瑞滨通过调节a）肿瘤表型、B）细胞因子/趋化因子环境和c）促凋亡/抗凋亡基因比率增加了对穿孔素/颗粒酶介导的CTL杀伤的敏感性。这里提供的数据提出了一个复杂的机制，这是不同的和互补的免疫原性细胞死亡。这种分子特征可能有助于预测对免疫治疗的反应，并为疫苗与顺铂/长春瑞滨方案联合使用的潜在临床获益提供依据。

Certain chemotherapeutic regimens trigger cancer cell death while inducing dendritic cell maturation and subsequent immune responses. However, chemotherapy-induced immunogenic cell death (ICD) has thus far been restricted to select agents. In contrast, several chemotherapeutic drugs modulate antitumor immune responses, despite not inducing classic ICD. In addition, in many cases tumor cells do not die after treatment. Here, using docetaxel, one of the most widely used cancer chemotherapeutic agents, as a model, we examined phenotypic and functional consequences of tumor cells that do not die from immunogenic cell death. Docetaxel treatment of tumor cells did not induce ATP or HMGB1 secretion, or cell death. However, calreticulin exposure was observed in all cell lines examined after chemotherapy treatment. Killing by CEA, MUC-1, or PSA-specific CD8+ CTLs was significantly enhanced after docetaxel treatment. This killing was associated with increases in components of antigen-processing machinery, and mediated largely by calreticulin membrane translocation, as determined by functional knockdown of calreticulin, PERK, or calreticulin-blocking peptide. A docetaxel-resistant cell line was selected (MDR-1+, CD133+) by continuous exposure to docetaxel. These cells, while resistant to direct cytostatic effects of docetaxel, were not resistant to the chemomodulatory effects that resulted in enhancement of CTL killing. We provided an operational definition of "immunogenic modulation," where exposure of tumor cells to nonlethal/sublethal doses of chemotherapy alters tumor phenotype to render the tumor more sensitive to CTL killing. These observations are distinct and complementary to immunogenic cell death and highlight a mechanism whereby chemotherapy can be used in combination with immunotherapy. Chemotherapy with platinum doublets, including cisplatin plus vinorelbine, is standard of care for non-small cell lung cancer (NSCLC). Sublethal exposure to certain chemotherapeutic agents has been demonstrated to alter the phenotype or biology of human tumor cells, rendering them more susceptible to cytotoxic T lymphocyte (CTL)-mediated lysis. However, the effects of cisplatin/vinorelbine on tumor sensitivity to T-cell cytotoxicity and its molecular mechanisms have not been fully elucidated. Here, we examined the effect of this chemotherapy on growth, cell-surface phenotype, and CTL-mediated lysis of five distinct human lung carcinoma cell lines in vitro, and examined the molecular mechanisms associated with enhanced CTL sensitivity. These studies demonstrate that sublethal exposure of human lung tumor cells to the platinum doublet modulates tumor cell phenotype, and increases sensitivity to MHC-restricted perforin/granzyme-mediated CTL killing. These studies also demonstrate that exposure to chemotherapy markedly decreased the protein secretion ratio of TGF-beta/IL-8. We examined the gene expression profile of two lung tumor cell lines in order to identify a shared gene signature in response to sublethal cisplatin/vinorelbine and found coordinate expression of only 16 transcripts, including those for cytokine/chemokine expression and apoptosis such as TNF-alpha, IL8, CXCL5, and BCL-2 like genes. Overall, these results suggest that sublethal exposure to cisplatin/vinorelbine increases sensitivity to perforin/granzyme-mediated CTL killing by modulation of a) tumor phenotype, b) cytokine/chemokine milieu, and c) the pro-apoptotic/anti-apoptotic gene ratio. The data presented here propose a complex mechanism that is distinct from and complementary to that of immunogenic cell death. This molecular signature may be useful in predicting responses to immunotherapy as well as provide the rationale for the potential clinical benefit of the combined use of vaccine with cisplatin/vinorelbine regimens.