Validation of Coccidiodes Target Antigens for Immunotherapy

我们已经证明，我们可以逆转播散性淋巴瘤患者淋巴细胞中的抗原特异性无反应性， 球孢子菌感染的树突状细胞（DC）。绵羊小球体裂解物。一 观察到响应于小球裂解物的Thl细胞因子谱和细胞增殖。以来 小球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的DC系统中的重组抗原，这些抗原已被证明具有保护作用。 在小鼠模型中对抗球孢子菌属真菌的致死攻击。我们假设直流电脉冲 在佐剂存在下，来自球孢子菌球的确定的重组抗原将引发保护性免疫应答。 播散性肺炎患者淋巴细胞中TH 1细胞因子谱和逆转球孢子虫无能 球孢子菌病检验这一假设的具体目的是验证重组球孢子虫抗原， Ag 2/PRA（富含脯氨酸的抗原）、CSA（球孢子菌特异性抗原）和Ag 2/PRA-CSA嵌合融合体 通过评估来自nafve，健康免疫供体和来自患有nafve的患者的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，在存在和不存在单磷酰脂质A（MPL）佐剂的情况下， 将分析对抗原脉冲的DC应答的淋巴细胞亚群的细胞因子分泌和细胞增殖。 表面标记。DC上的趋化因子受体以及中枢和效应记忆淋巴细胞标志物 将通过流式细胞术进行分析。从这些重组抗原研究中获得的数据将 评价并与T27 K球孢子菌球溶胞产物进行比较，以确定 淋巴细胞反应符合保护作用。这个项目的总体目标是定位我们的临床试验 使用重组球虫抗原的免疫疗法治疗播散性 球孢子菌病是折射到抗真菌治疗。

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.