CTL DEFINED ANTIGENS IN NON SMALL CELL LUNG CANCER

非小细胞肺癌（NSCLC）是世界各地女性和女性的主要杀手。NSCLC患者需要额外的治疗选择。该提案旨在研究针对NSCLC的细胞毒性T淋巴细胞（CTL）应答，以开发新的免疫选择。该提案提出的问题将定义和表征NSCLC特异性CTL识别的抗原。它还将研究一种将NSCLC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抑制剂在NSCLC中的临床试验。提出了四个具体的研究目标。前两个特异性目的将鉴定HLA-A2限制性NSCLC特异性CTL以及鉴定和表征被识别的抗原。在特定目标3中，我们将研究从HLA-A3开始的其他限制性元件在NSCLC肽呈递中的使用。NSCLC特异性CTL的鉴定将涉及使用体外共培养方案（混合淋巴细胞肿瘤细胞培养物，MLTC），将从正常供体和NSCLC患者的外周血（PBMC）获得的淋巴细胞与HLA-A基因座匹配的同种异体长期NSCLC肿瘤细胞系组合。NSCLC细胞系经基因修饰以表达淋巴细胞共刺激分子CD 80（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呈递细胞。将使用分子生物学技术和现有基因克隆策略鉴定特异性目标2中CTL定义的抗原。虽然我们目前的初步数据表明，我们可以成功地产生HLA-A2特异性CTL使用所描述的系统，具体目标4建议研究一种替代的，可能更有效的手段，NSCLC抗原呈递。提出了NSCLC肿瘤细胞系和树突状细胞之间的融合。所得体细胞杂交体将用于体外产生特异性CTL。这些在NSCLC中产生特异性CTL的尝试将大大增加我们对这种疾病的免疫应答的知识基础，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.