Biologic Therapy for B-cell non-Hodgkin's Lymphoma

描述（由申请人提供）：B细胞非霍奇金淋巴瘤（NHL）是美国癌症相关死亡的第六大常见原因。虽然许多侵袭性淋巴瘤患者可以用细胞毒性疗法治愈，但大多数惰性淋巴瘤用目前的疗法无法治愈。因此，需要新的有效疗法来治疗这些患者。我们研究的目标是通过利用增强对恶性B细胞的免疫反应的策略，为B细胞NHL患者开发新的生物疗法。在先前的资助期间，我们评估了在B细胞淋巴瘤患者中加入IL-12（一种免疫刺激细胞因子）是否会增加利妥昔单抗的抗体依赖性细胞毒性。在临床试验中，我们发现IL-12没有显著增加反应率，高于单独使用利妥昔单抗的预期。在相关研究中，我们发现IL-12导致外周血细胞中基因的上调，但在肿瘤中没有类似的作用。在实验室研究中，我们发现效应细胞对IL-12免疫刺激反应的缺乏部分与具有抑制功能的肿瘤内T细胞的存在有关。最近的研究表明，CD 4 + CD 25+调节性T（Treg）细胞通过诱导对肿瘤特异性抗原的外周耐受而参与抗肿瘤免疫的调节。然而，关于Treg细胞对B细胞NHL中的肿瘤特异性T细胞免疫以及随后对恶性B细胞生长的影响的数据很少。在初步研究中，我们已经确定了一个亚组的CD 4 + CD 25 + T细胞与Treg细胞表型，是目前在B细胞NHL。此外，我们发现这些Treg细胞具有抑制B细胞NHL中肿瘤浸润性CD 4+和CD 8 + T细胞的能力，并且它们响应于恶性B细胞产生的趋化因子如CCL 22而迁移。我们的中心假设是肿瘤Treg细胞通过抑制肿瘤浸润性T细胞促进恶性淋巴瘤B细胞的生长，并且恶性B细胞通过选择性地将Treg细胞募集到B细胞NHL的区域而发挥积极作用。因此，我们建议首先确定这些Treg细胞被募集到非霍奇金淋巴瘤中的恶性B细胞微环境的机制，并发现它们在肿瘤微环境中存在时是否获得抑制活性（Aim 1）。其次，我们将评估恶性B细胞是否与肿瘤微环境中的Treg细胞直接相互作用，从而协调对它们存在的耐受性（目的2）。第三，我们将确定肿瘤内Treg细胞的消耗和恶性B细胞的抑制以减少Treg细胞募集是否会导致B细胞NHL患者的临床益处（目的3）。我们预计，拟议的研究将提供一个更好地了解Treg细胞介导的影响，在B细胞恶性肿瘤。我们还预期，地尼白细胞介素2和白喉毒素融合蛋白denileukin diftitox与抗CD 20单克隆抗体利妥昔单抗的组合的临床使用将抑制B细胞淋巴瘤患者中的Treg细胞，并且还将耗尽恶性淋巴结中的淋巴瘤B细胞，从而防止Treg细胞进一步募集到B细胞淋巴瘤区域中。这种治疗组合将导致一种新的治疗方法来调节Treg细胞，这将导致B细胞NHL患者的临床获益。

DESCRIPTION (provided by applicant): B-cell non-Hodgkin lymphomas (NHL) are the sixth most common cause of cancer-related deaths in the United States. While many patients with aggressive lymphomas may be cured with cytotoxic therapy, most indolent lymphomas are incurable with current therapy. Novel effective therapies are therefore needed to treat these patients. The goal of our research is to develop novel biologic therapies for patients with B-cell NHL by utilizing strategies that augment the immune response to the malignant B-cell. During the previously funded period we evaluated whether adding IL-12, an immunostimulatory cytokine, to rituximab in patients with B-cell lymphoma would increase the antibody dependent cytotoxicity of rituximab. In the clinical trial, we found that IL-12 did not significantly increase the response rate above what would be expected with rituximab alone. In correlative studies we found that IL-12 resulted in upregulation of genes in peripheral blood cells but did not have similar effects in the tumor. In laboratory studies, we found that the lack of effector cell response to immune stimulation with IL-12 was related in part to the presence of intratumoral T-cells with suppressive function. Recent studies have suggested that CD4+CD25+ regulatory T (Treg) cells are involved in the regulation of anti-tumor immunity by inducing peripheral tolerance to tumor specific antigens. However, there are little data regarding the effect of Treg cells on tumor-specific T cell immunity in B-cell NHL and subsequently on the malignant B-cell growth. In preliminary studies, we have identified a subset of CD4+CD25+ T cells with a Treg cell phenotype that are present in B-cell NHL. In addition, we find that these Treg cells have the ability to suppress tumor-infiltrating CD4+ and CD8+ T cells in B-cell NHL and that they migrate in response to chemokines such as CCL22 produced by the malignant B-cells. Our central hypothesis is that tumor Treg cells contribute to the growth of malignant lymphoma B cells by suppressing tumor-infiltrating T cells and that malignant B-cells play an active role by selectively recruiting Treg cells to the areas of B-cell NHL. We therefore propose to firstly determine the mechanism by which these Treg cells are recruited to the malignant B-cell microenvironment in non-Hodgkin lymphoma and to discover whether they gain suppressive activity when present in the tumor microenvironment (Aim 1). Secondly, we will assess whether malignant B-cells interact directly with Treg cells in the tumor microenvironment and thereby orchestrate tolerance to their presence (Aim 2). Thirdly, we will establish whether depletion of intratumoral Treg cells, and inhibition of malignant B-cells to decrease Treg cell recruitment, will result in clinical benefit for patients with B-cell NHL (Aim 3). We anticipate that the proposed research will provide a better understanding of the Treg cell-mediated effects in B-cell malignancies. We also anticipate that the clinical use of denileukin diftitox, an interleukin-2 and diphtheria toxin fusion protein, in combination with rituximab, an anti-CD20 monoclonal antibody, will inhibit Treg cells in B-cell lymphoma patients and will also deplete lymphoma B-cells in malignant lymph nodes thereby preventing further recruitment of Treg cells into areas of B-cell lymphoma. This treatment combination will lead to a novel therapeutic approach to modulating Treg cells that will result in clinical benefit for patients with B-cell NHL.