T CELL ACTIVATION FOR CANCER IMMUNOTHERAPY

T 细胞激活用于癌症免疫治疗

• 批准号: 6173794
• 负责人: ALFRED E CHANG
• 金额: $ 28.67万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6173794
• 关键词: CD28 molecule; CD3 molecule; T lymphocyte; animal tissue; antitumor antibody; cellular immunity; cytokine; cytotoxic T lymphocyte; growth factor receptors; helper T lymphocyte; immunocytochemistry; laboratory mouse; leukocyte activation /transformation; lymph nodes; monoclonal antibody; neoplasm /cancer immunotherapy; neoplastic transformation; passive immunization; protein protein interaction; receptor binding; tissue /cell culture; tumor necrosis factor alpha

Our laboratory has focused on the generation of tumor reactive T cells derived from lymph nodes (LNs) primed by progressive tumors or by tumor vaccinations. Based upon extensive animal studies, we have shown that these tumor-primed LNs can be secondarily activated in vitro by anti-CD3 mAb, as a surrogate antigen, and expanded in IL-2 for subsequent adoptive immunotherapy. These anti-CD3 activated cells mediate immunologically specific tumor regression. This culture method preferentially activates CD8+ cells which have been found to mediate the antitumor responses in vivo; without the requirement of CD4+ cells. These effector cells appear to manifest their antitumor effects by the release of type 1 cytokines (ie. IFNgamma and GM-CSF) in response to tumor antigen. By contrast, effector cells which release type 2 cytokines (ie., IL-10) do not mediate tumor regression in vivo. More recently, we have found that we can further stimulate tumor- primed LN cells with the addition of anti-CD28 mAb as a co- stimulatory signal. This has resulted in enhanced cytokine release in response to tumor antigen. In addition, we have observed the preferential activation of CD4+ tumor reactive T cells utilizing this culture procedure when purified CD3+ cells are activated as opposed to the unfractionated LN population. These findings indicate at the components of the cell population and the in vitro activating conditions can have a profound effect on the subsequent cell population which is generated. Our preliminary results provide us with methods to examine the role of CD4 versus CD8+ T cells in adoptive immunotherapy; and to determine if enriched subpopulations of T cells with tumor reactivity can be selectively cultured. We propose the following specific aims: l) To examine the antitumor reactivity of tumor-primed LN cells after in vitro activation with anti-CD3 mAb and co-stimulation with anti-CD28 mAb; 2)To examine the antitumor reactivity of tumor-primed LN cells after in vitro activation with anti-Vbeta mAbs and co-stimulation with anti-CD28 mAb; 3) To examine the effect of different cytokines during the in vitro activation of tumor-primed LN cells on the maturation of pre- effector cells and; 4) To examine other co-stimulatory signals on the in vitro activation of tumor-primed LN cells (i.e. anti-CD40 or anti-41BB). The objectives of these aims are to provide us with a better understanding regarding the cellular effector mechanisms involved in the immune destruction of tumor, and to selectively activate and expand pre-existing tumor-reactive T cells present at low frequencies.

我们的实验室专注于从进展性肿瘤或肿瘤疫苗启动的淋巴结(LN)中产生肿瘤反应性T细胞。基于广泛的动物研究，我们已经证明这些肿瘤诱导的LN可以在体外被抗CD3单抗作为替代抗原二次激活，并在IL-2中扩增，以用于后续的过继免疫治疗。这些抗CD3激活的细胞介导免疫特异性肿瘤消退。这种培养方法优先激活体内已发现的介导抗肿瘤反应的CD8+细胞，而不需要CD4+细胞。这些效应细胞似乎通过释放1型细胞因子(即。干扰素和GM-CSF)来应答肿瘤抗原。相比之下，释放2型细胞因子(即IL-10)的效应细胞在体内并不介导肿瘤的消退。最近，我们发现通过添加抗CD28单抗作为共刺激信号，我们可以进一步刺激肿瘤诱导的LN细胞。这导致了对肿瘤抗原反应的细胞因子释放增强。此外，我们观察到，当纯化的CD3+细胞被激活时，利用这种培养程序优先激活CD4+肿瘤反应性T细胞，而不是未分离的LN群体。这些发现表明，细胞群体的组成和体外激活条件可以对随后产生的细胞群体产生深远的影响。我们的初步结果为我们提供了检测CD4和CD8+T细胞在过继免疫治疗中的作用的方法；并确定是否可以选择性培养具有肿瘤反应性的丰富的T细胞亚群。我们提出的具体目标如下：L)检测肿瘤诱导的LN细胞经抗CD3mAb体外激活和抗CD28mAb共刺激后的抗肿瘤活性；2)检测肿瘤诱导的LN细胞经抗Vbeta单抗体外激活和抗CD28mAb共同刺激后的抗肿瘤反应性；3)检测肿瘤诱导的LN细胞体外激活过程中不同细胞因子对前效应细胞成熟的影响；4)检测肿瘤诱导的LN细胞体外激活的其他共刺激信号(如抗CD40或抗41BB)。这些目标的目的是让我们更好地了解参与肿瘤免疫破坏的细胞效应机制，并选择性地激活和扩大现有的低频率的肿瘤反应性T细胞。