IL-1 HOMOLOGUES PROMOTE THE ACUTE INFLAMMATORY RESPONSE

我们现在已经确定了扩展的IL-1家族[IL-1Fx]的10个不同的成员，其中包括IL-18作为最亲缘关系的成员。其中许多细胞因子的生物学作用尚不清楚，但IL-1α/IL-1F1、IL-1β/IL-1F2、IL-18/IL-1F4和IL-1H4/IL-1F7用作基因构建物时可促进抗肿瘤反应。我们先前在体外已经证明，rIL-18在小鼠肿瘤模型中的抗肿瘤作用可以与小鼠DC、NK细胞、肿瘤、T细胞和IL-18共培养模拟，导致肿瘤特异性T细胞的产生。IL-18的基因治疗依赖于DC和NK细胞、Fas信号，而不依赖于NKT细胞、IFNGamma或IL-12。一种新的IL-1同系物IL-1H4/IL-1F7也介导了抗肿瘤作用 IL-12依赖和Fas依赖的效应，显示了可归因于IL-12和IL-18的混合生物效应。我们现在将在人类体外研究中探索这些细胞因子，DC和NK细胞相互作用，以促进T细胞对人类黑色素瘤的反应。因此，我们将探索其他IL-1同源物，包括IL-1α、IL-1β、IL-1RA、IL-1F5/IL-1F9和IL-1H4[IL-1F7]在体外培养中是否介导相似或不同的作用。我们将评估人类黑色素瘤体外培养中共培养系统的关键组成部分。我们还将评估IL-12的产生是否以依赖于IFN的方式在NK中成功地产生特定的T细胞的培养中得到增强 DC与肿瘤共培养。我们将与项目II合作，比较IL-1家族成员的作用，以评估小鼠黑色素瘤的类似读数，以及项目II中肿瘤坏死因子家族成员和扩展的IL-12家族成员的相对贡献作用。一个重要的问题将是在黑色素瘤患者体内测试这些前提，我们将在评估多发性皮肤黑色素瘤转移患者的局部肿瘤反应的创新临床方案中进行评估，并在项目II中与黑色素瘤患者的体外策略进行对比。我们将在以下具体目标中测试这些概念：目的1.评估IL-1同源物及其促进NK和DC激活、极化的能力；目的2.体外用IL-1同源物建立对自体和MHC相合的黑色素瘤的特异性T细胞反应性；目的3.将NK和DC生物标志物与IL-1同源物和蛋白质组学检测整合在治疗黑色素瘤患者中；目的4.在序贯临床试验中对黑色素瘤患者进行ANK/DC治疗。

We now have ten separate members of the extended IL-1 family [IL-1Fx] identified including IL-18 as the most distantly related member. The biologic role of many of these cytokines is unclear but IL-1alpha/IL-1F1, IL-1beta/IL-1F2, IL-18/IL-1F4, and IL-1H4/IL-1F7 promote antitumor responses when used as genetic constructs. We have previously shown in vitro that the antitumor effects of rIL-18 delivered in murine tumor models can be mimicked with coculture of murine DCs, NK cells, tumor, T-cells and IL-18, leading to the generation of tumor specific T-cells. IL-18 gene therapy is dependent on DC and NK cells, Fas signaling but not on NKT cells nor IFNgamma nor IL-12. A novel IL-1 homologue, IL-1H4/IL-1F7 also mediates antitumor effects, which are IL-12-dependent and Fas dependent, demonstrating a mixture of biologic effects attributable to IL-12 and IL-18. We will now explore these cytokines in human in vitro studies with DC and NK cells interacting to promote a T-cell response to human melanoma. We will thus explore whether the other IL-1 homologues including IL-1alpha, IL-1beta, IL1RA, IL-1F5/IL-1F9 and IL-1H4 [IL-1F7] mediate similar or dissimilar effects in in vitro culture. We will evaluate the critical components of the coculture system in human in vitro cultures with human melanoma. We will also evaluate whether IL-12 production is enhanced in an IFNgamma dependent manner in such cultures generating successful development of specific T-cells in NK and DC cultured with tumor. We will contrast the role of the IL-1 family members in cooperation with Project I to assess similar readouts in murine melanoma and the relative contributory roles of the TNF family members and the extended IL-12 family members with Project II. An important question will be to test these premises in vivo in patients with melanoma and we will assess this in innovative clinical protocols assessing local tumor response in patients with multiple cutaneous melanoma metastases and contrast with ex vivo strategies in patients with melanoma in Project II. We will test these notions in the following Specific Aims: Aim 1. Evaluate IL-1 homologues and their ability to promote NK and DC activation, polarization; Aim 2. Develop specific T-cell reactivity to autologous and MHC matched melanoma in vitro with IL-1 homologues; Aim 3. Integrate NK and DC biomarkers with IL-1 homologues and proteomic assays in treated melanoma patients and Aim 4. Administer ANK/DC to melanoma patients in sequential clinical trials.