Validation of Coccidiodes Target Antigens for Immunotherapy

我们已经证明，我们可以逆转播散性疾病患者淋巴细胞中的抗原特异性无能。 用球虫球体裂解物致敏的树突状细胞(DC)孵育球孢子菌病。一个 观察小球裂解物对Th1细胞因子谱和细胞增殖的影响。自.以来 球体裂解物不太可能被批准作为临床使用的抗原制剂，总体目标是 这个项目的目的是验证我们DC系统中已被证明具有保护性的重组抗原 在小鼠模型中对抗球虫类真菌的致死攻击。我们假设DC脉冲波 球虫球虫小球在佐剂存在下定义的重组抗原将引起保护性 播散性疾病患者淋巴细胞Th1细胞因子与逆转球虫无能的关系 球孢子菌病。检验这一假设的具体目的是验证重组球虫抗原， AG2/PRA、CsA和AG2/PRA-CsA嵌合融合 通过评估来自NAFVE、健康免疫捐赠者和慢性粒细胞白血病患者的DC表型和功能 播散性球孢子菌病。在确定重组抗原如何影响DC生成的同时， 表型和功能，在有或没有单磷脂A(MPL)佐剂的情况下， 对抗原冲击的DC有反应的淋巴细胞亚群将被分析细胞因子的分泌和细胞 曲面标记。树突状细胞表面趋化因子受体以及中枢和效应性记忆淋巴细胞标志物 将通过流式细胞仪进行分析。从这些重组抗原研究中获得的数据将是 评估并与T27K球虫球体裂解物进行比较，以确定是否 淋巴细胞反应符合保护性特征。这个项目的总体目标是为我们的临床试验定位 重组球虫抗原免疫治疗在播散性疾病中的应用 对抗真菌治疗屈光的球孢子菌病。

We have shown that we can reverse antigen-specific anergy in lymphocytes from patients with disseminated coccidioidomycosis by incubating them with dendritic cells (DC) pulsed with C. immitis spherule lysate. A Thl cytokine profile and cellular proliferation were observed in response to the spherule lysate. Since spherule lysates are unlikely to be approved as an antigen preparation for clinical use, the overall objectives of this project are to validate recombinant antigens in our DC system that have been shown to be protective in murine models against lethal challenge with coccidioides fungus. We hypothesize that DC pulsed with defined recombinant antigens from Coccidioides spherules in the presence of adjuvant will elicit a protective TH1 cytokine profile and reverse coccidioidal anergy in lymphocytes from patients with disseminated coccidioidomycosis. Specific aims to test this hypothesis are to validate recombinant coccidioides antigens, Ag2/PRA (proline rich antigen), CSA (coccidioides specific antigen) and Ag2/PRA-CSA chimeric fusion protein by evaluating DC phenotype and function from nafve, healthy immune donors and from patients with disseminated coccidioidomycosis. While determining how recombinant antigens affect DC generation, phenotype and function, in the presence and absence of monophosphoryl Lipid A (MPL) adjuvant, lymphocyte subsets that respond to antigen-pulsed DC will be analyzed for cytokines secretion and cell surface markers. Chemokine receptors on DC as well as central and effector memory lymphocyte markers will be analyzed by flow cytometry. Data obtained from these studies with recombinant antigens will be evaluated and compared with that obtained with T27K coccidioides spherule lysate to determine if the lymphocyte responses fit a protective profile. The overall goal of this project is to position us for a clinical trial using immunotherapy with recombinant coccidoides antigens for the treatment of disseminated coccidioidomycosis that is refractive to anti-fungal treatment.