Butterfield Lisa H, Buffo Mary Jo
University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Methods Mol Biol. 2014;1102:71-82. doi: 10.1007/978-1-62703-727-3_5.
Cancer vaccines are designed to activate an immune response to tumor-specific or tumor-associated antigens expressed by the tumor. Cancer vaccines take many forms, including synthetic peptides, tumor cells and lysates, cell lines, and autologous antigen presenting cells like dendritic cells. The target antigens may be known, or "defined" in the vaccine, or unknown. In melanoma, more so than in other cancers, a large number of immunogenic "shared" antigens (tumor-specific or tumor-associated) have been identified. This allows for vaccination of groups of patients with the same vaccine, and also allows for testing for melanoma tumor immunity even when the vaccine does not include defined antigens. For the cancer vaccine field, the goal of a prognostic or predictive biomarker has yet to be achieved. However, the primary immunologic goal of any cancer vaccine is the induction (or amplification) of an immune response against the tumor, therefore the primary goal of immunologic monitoring in this setting, is testing for that response. In this chapter, we present standardized methodology from a central immunologic monitoring laboratory for melanoma cancer vaccine immune response assessment by the Enzyme-Linked Immunosorbant Spot (ELISPOT) assay. This assay allows for enumeration of antigen-specific cells in a plate format. We present the Interferon (IFN)-γ-producing lymphocyte assay, but the platform is easily adjusted to several cell types and several secreted molecules.
癌症疫苗旨在激活针对肿瘤表达的肿瘤特异性或肿瘤相关抗原的免疫反应。癌症疫苗有多种形式,包括合成肽、肿瘤细胞和裂解物、细胞系以及自体抗原呈递细胞(如树突状细胞)。疫苗中的靶抗原可能是已知的,即“明确的”,也可能是未知的。与其他癌症相比,黑色素瘤中已鉴定出大量免疫原性“共享”抗原(肿瘤特异性或肿瘤相关)。这使得可以用相同的疫苗对一组患者进行接种,并且即使疫苗不包含明确的抗原,也可以检测黑色素瘤肿瘤免疫。对于癌症疫苗领域而言,预后或预测性生物标志物的目标尚未实现。然而,任何癌症疫苗的主要免疫学目标都是诱导(或增强)针对肿瘤的免疫反应,因此在这种情况下免疫监测的主要目标是检测这种反应。在本章中,我们介绍了一个中央免疫监测实验室用于通过酶联免疫斑点(ELISPOT)测定评估黑色素瘤癌症疫苗免疫反应的标准化方法。该测定允许以平板形式计数抗原特异性细胞。我们介绍了产生干扰素(IFN)-γ的淋巴细胞测定,但该平台可轻松调整以适用于多种细胞类型和多种分泌分子。