Vaccine and Drug Combination Therapy for Human Cancers

某些化疗方案在诱导树突状细胞的同时触发癌细胞死亡， 细胞成熟和随后的免疫反应。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试剂。相比之下， 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， 作为模型，我们检查了最广泛使用的癌症化疗剂的表型和 肿瘤细胞的功能性后果不会死于免疫原性细胞死亡。多西他 肿瘤细胞的处理不诱导ATP或HMGB 1分泌或细胞死亡。然而，在这方面， 在化疗处理后检查的所有细胞系中观察到钙网蛋白暴露。 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。 治疗这种杀伤作用与抗原加工成分的增加有关。 机械，并主要由钙网蛋白膜易位介导，如由 钙网蛋白、PERK或钙网蛋白阻断肽的功能性敲低。一 通过连续暴露于紫杉醇，选择紫杉醇耐药细胞系（MDR-1+，CD 133+）。 多西他赛。这些细胞虽然对多西他赛的直接细胞生长抑制作用有抗性，但对多西他赛的细胞生长抑制作用没有影响。 对导致CTL杀伤增强的化学调节作用具有抗性。我们 提供了“免疫原性调节”的操作定义，其中肿瘤细胞的暴露 非致死/亚致死剂量的化疗改变了肿瘤表型， 对CTL杀伤敏感。这些观察结果与免疫原性 细胞死亡，并强调了一种机制，化疗可以与 免疫疗法铂双联化疗，包括顺铂加长春瑞滨， 非小细胞肺癌（NSCLC）的标准治疗。亚致死暴露于某些 已经证明化疗剂改变人肿瘤的表型或生物学 细胞，使它们更容易受到细胞毒性T淋巴细胞（CTL）介导的裂解。 然而，顺铂/长春瑞滨对肿瘤对T细胞毒性的敏感性的影响， 其分子机制尚未完全阐明。在这里，我们研究了这一点的影响， 化疗对生长、细胞表面表型和CTL介导的五种不同人类肿瘤细胞溶解的影响 肺癌细胞株，并研究了与肺癌相关的分子机制。 增强CTL敏感性。这些研究表明，人类肺部的亚致死暴露 肿瘤细胞与铂双联体的结合调节肿瘤细胞表型，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， 暴露于化疗显著降低TGF-β/IL-8的蛋白分泌比率。我们 检测了两种肺肿瘤细胞系的基因表达谱， 对亚致死性顺铂/长春瑞滨的应答共享基因签名， 仅表达16种转录本，包括细胞因子/趋化因子表达的转录本， 细胞凋亡，如TNF-α、IL 8、CXCL 5和BCL-2样基因。总的来说，这些结果表明， 顺铂/长春瑞滨的亚致死暴露增加了对 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。数据 这里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 免疫疗法以及提供的潜在临床效益的理由， 联合使用疫苗与顺铂/长春瑞滨方案。

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.