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.

我们的实验室集中于产生源自淋巴结（LNS）的肿瘤反应性T细胞，该细胞由进行性肿瘤或肿瘤疫苗接种。基于广泛的动物研究，我们已经表明，这些肿瘤植入的LN可以第二次通过抗CD3 mAb作为替代抗原在体外激活，并在IL-2中扩展，以进行随后的采用免疫疗法。这些抗CD3激活的细胞介导免疫学特异性的肿瘤消退。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的1型细胞因子（即ifngamma和GM-CSF）的释放来表现其抗肿瘤作用。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的消退。最近，我们发现，我们可以通过添加抗CD28 mAb作为共刺激信号来进一步刺激肿瘤的LN细胞。这导致响应肿瘤抗原的细胞因子释放增强。此外，当纯化的CD3+细胞被激活而不是未分离的LN群体相反，我们已经观察到使用该培养程序的CD4+肿瘤反应性T细胞的优先激活利用这种培养程序。这些发现表明在细胞群的成分上，体外激活条件可能会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细胞在收养免疫疗法中的作用的方法。并确定是否可以选择性地培养具有肿瘤反应性的T细胞的富集亚群。我们提出以下特定目的：l）检查抗CD3 mAb体外激活后肿瘤引发的LN细胞的抗肿瘤反应性，并与抗CD28 MAB共刺激； 2）检查用抗VBETA mAb体外激活肿瘤引发的LN细胞的抗肿瘤反应性，并与抗CD28 MAB共同刺激； 3）检查在肿瘤培养的LN细胞体外激活过程中不同细胞因子对效应细胞成熟的影响； 4）检查有关肿瘤培养的LN细胞体外激活（即抗CD40或抗41BB）的其他共刺激信号。这些目的的目的是为我们提供有关肿瘤免疫破坏的细胞效应机制的更好理解，并在低频下选择性地激活和扩展存在先前存在的肿瘤反应性T细胞。