IL-1 HOMOLOGUES PROMOTE THE ACUTE INFLAMMATORY RESPONSE

我们现在有10个独立的成员的扩展IL-1家族[IL-1Fx]确定，包括IL-18作为最远的相关成员。许多这些细胞因子的生物学作用尚不清楚，但IL-1 α/IL-1F 1、IL-1 β/IL-1F 2、IL-18/IL-1F 4和IL-1H 4/IL-1F 7在用作遗传构建体时可促进抗肿瘤反应。我们先前已经在体外证明，在鼠肿瘤模型中递送的rIL-18的抗肿瘤作用可以用鼠DC、NK细胞、肿瘤、T细胞和IL-18的共培养物来模拟，导致肿瘤特异性T细胞的产生。IL-18基因治疗依赖于DC和NK细胞、Fas信号传导，但不依赖于NKT细胞、IFN γ或IL-12。一种新的IL-1同源物IL-1H 4/IL-1F 7也介导抗肿瘤作用 IL-12依赖性和Fas依赖性的生物学效应，证明了IL-12和IL-18的生物学效应的混合。我们现在将在体外研究中探索这些细胞因子，DC和NK细胞相互作用促进T细胞对人黑色素瘤的反应。因此，我们将探讨其他IL-1同系物，包括IL-1 α，IL-1 β，IL-1 RA，IL-1F 5/IL-1F 9和IL-1H 4 [IL-1F 7]是否在体外培养中介导相似或不同的作用。我们将评估与人黑色素瘤体外培养的人共培养系统的关键组成部分。我们还将评估在NK细胞中成功产生特异性T细胞的培养物中，IL-12的产生是否以IFN γ依赖性方式增强。 DC与肿瘤共同培养。我们将对比IL-1家族成员与项目I合作的作用，以评估小鼠黑色素瘤中的类似读数，以及TNF家族成员和扩展的IL-12家族成员与项目II的相对贡献作用。一个重要的问题是在黑色素瘤患者体内测试这些前提，我们将在创新的临床方案中评估这一点，评估多发性皮肤黑色素瘤转移患者的局部肿瘤缓解，并与项目II中黑色素瘤患者的离体策略进行对比。我们将在以下具体目标中检验这些概念：目标1。评估IL-1同系物及其促进NK和DC活化、极化的能力;目的2.在体外用IL-1同源物开发对自体和MHC匹配的黑素瘤的特异性T细胞反应性;目的3。整合NK和DC生物标志物与IL-1同系物和蛋白质组学测定在治疗的黑色素瘤患者和目的4。在序贯临床试验中向黑色素瘤患者给予ANK/DC。

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.