CTL DEFINED ANTIGENS IN NON SMALL CELL LUNG CANCER

非小细胞肺癌(NSCLC)是世界上女性和女性的头号杀手。非小细胞肺癌患者需要更多的治疗选择。该建议旨在检测细胞毒性T淋巴细胞(CTL)对非小细胞肺癌的反应，以开发新的免疫治疗方案。该提案提出了一些问题，这些问题将定义和表征NSCLC特异性CTL识别的抗原。它还将研究一种将非小细胞肺癌抗原呈递给淋巴细胞前体的新方法。收集到的部分或全部信息可能会导致新型免疫治疗试剂在非小细胞肺癌中的临床试验。本文提出了四个具体的研究目标。前两个特异性目标将识别人类白细胞抗原A2限制性非小细胞肺癌特异性CTL，以及识别和表征被识别的抗原。在特定的目标3中，我们将研究从人类白细胞抗原-A3开始的其他限制元件在非小细胞肺癌多肽呈递中的使用。NSCLC特异性CTL的鉴定将涉及使用一种体外共培养方案(混合淋巴细胞肿瘤细胞培养，MLTC)，该方案将从正常人和NSCLC患者的外周血(PBMC)中获得的淋巴细胞与HLAA基因位点匹配的长期NSCLC肿瘤细胞株相结合。NSCLC系经过基因改造，可以表达淋巴细胞共刺激分子CD80(B7.1)。因此，携带I类MHC分子和潜在肿瘤抗原的肿瘤细胞将被制成有效的提呈细胞。将使用分子生物学技术和现有的基因克隆策略来鉴定特定目标2中CTL定义的抗原。虽然我们提供的初步数据表明，我们可以使用所描述的系统成功地产生人类白细胞抗原A2特异性CTL，但特殊目的4建议研究一种替代的、可能更有效的非小细胞肺癌抗原提呈方法。NSCLC肿瘤细胞系与树突状细胞之间的融合被提出。由此产生的体细胞杂交将用于在体外产生特定的CTL。这些在非小细胞肺癌中产生特异性CTL的尝试将极大地增加我们对这种疾病的免疫学反应的知识基础，并为疫苗或细胞免疫治疗方案提供关键试剂。

Non-small cell lung cancer (NSCLC), is a leading killer of both women and women around the world. Patients with NSCLC are in need of additional therapeutic options. This proposal seeks to examine the cytotoxic T lymphocyte (CTL) response against NSCLC in order to develop novel immunotherapeutic options. The proposal asks questions which will define and characterize the antigens recognized by NSCLC specific CTL. It also will investigate a novel means of presenting NSCLC antigens to lymphocyte precursors. Some or all of the information gleaned may lead to clinical trials of novel immunotherapeutic reagents in NSCLC. Four Specific aims are proposed for study. The first two Specific Aims will identify HLA- A2 restricted NSCLC specific CTL as well as identify and characterize the antigens being recognized. In Specific Aim 3, we will examine the use of other restriction elements beginning with HLA-A3, in the presentation of NSCLC peptides. The identification of NSCLC specific CTL will involve the use of an in vitro co- culture scheme (Mixed lymphocyte tumor cell culture, MLTC) combining lymphocytes obtained from peripheral blood (PBMC) of normal donors and NSCLC patients with HLA-A locus matched allogeneic long term NSCLC tumor cell lines. The NSCLC lines are gene modified to express the lymphocyte co-stimulatory molecule, CD80 (B7.1). Thus, the tumor cells bearing class I MHC molecules and potential tumor antigens will be made into efficient presenting cells. Identification of CTL defined antigens in Specific Aim 2 will be done using molecular biologic techniques and existing gene- cloning strategies. While we present preliminary data showing that we can successfully generate HLA-A2 specific CTL using the described system, Specific Aim 4 proposes to study an alternative and possibly more efficient means of NSCLC antigen presentation. Fusions are proposed between NSCLC tumor cell lines and dendritic cells. Resultant somatic cell hybrids will be used to generate specific CTL in vitro. These attempts to generate specific CTL in NSCLC will add greatly to our knowledge base of the immunological response to this disease as well as provide critical reagents for vaccine or cellular immunotherapy protocols.