T CELL ACTIVATION FOR CANCER IMMUNOTHERAPY

T 细胞激活用于癌症免疫治疗

基本信息

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
项目类别：
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

项目摘要

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淋巴结 (LN) 中生成肿瘤反应性 T 细胞，这些淋巴结是由进展性肿瘤或肿瘤疫苗接种引发的。基于广泛的动物研究，我们表明这些肿瘤引发的 LN 可以在体外被抗 CD3 mAb 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 激活细胞介导免疫特异性肿瘤消退。这种培养方法优先激活 CD8+ 细胞，这些细胞已被发现可介导体内抗肿瘤反应；无需 CD4+ 细胞。这些效应细胞似乎通过响应肿瘤抗原释放 1 型细胞因子（即 IFNγ 和 GM-CSF）来表现出其抗肿瘤作用。相比之下，释放2型细胞因子（即IL-10）的效应细胞不会介导体内肿瘤消退。最近，我们发现通过添加抗 CD28 mAb 作为共刺激信号，可以进一步刺激肿瘤引发的 LN 细胞。这导致响应肿瘤抗原的细胞因子释放增强。此外，我们还观察到，与未分级的 LN 群体相比，当纯化的 CD3+ 细胞被激活时，利用该培养程序优先激活 CD4+ 肿瘤反应性 T 细胞。这些发现表明细胞群的组成和体外激活条件可以对随后产生的细胞群产生深远的影响。我们的初步结果为我们提供了检查 CD4 与 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富集 T 细胞亚群。我们提出以下具体目标： l) 检查肿瘤引发的 LN 细胞在体外用抗 CD3 mAb 激活并与抗 CD28 mAb 共刺激后的抗肿瘤反应性； 2)检测肿瘤引发的LN细胞在用抗Vbeta mAb体外激活并与抗CD28 mAb共刺激后的抗肿瘤反应性； 3) 检测肿瘤引发的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) 检查肿瘤引发的 LN 细胞体外激活的其他共刺激信号（即抗 CD40 或抗 41BB）。这些目标的目的是让我们更好地了解肿瘤免疫破坏所涉及的细胞效应机制，并选择性地激活和扩展低频率存在的现有肿瘤反应性 T 细胞。