CELLULAR HOST DEFENSES IN COCCIDIOIDOMYCOSIS

球孢子菌病中的细胞宿主防御

• 批准号: 2457742
• 负责人: D MITCHELL MAGEE
• 金额: $ 19.13万
• 依托单位: TEXAS CENTER FOR INFECTIOUS DISEASE
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2457742
• 关键词: Coccidioides immitis; cell type; cellular immunity; coccidioidomycosis; cytokine; delayed hypersensitivity; enzyme linked immunosorbent assay; fungal vaccines; genetic strain; host organism interaction; interferon gamma; interleukin 12; interleukin 2; interleukin 4; laboratory mouse; liver cells; lung; lymph nodes; neutralizing antibody; polymerase chain reaction; spleen

DESCRIPTION (Adapted from applicant abstract): Host defenses against C. immitis are mediated by specifically sensitized T cells. In the mouse model, both genetically susceptible (BALB/c) and genetically resistant (DBA/2) mice mount cell-mediated immunity (CMI), as measured by delayed- type hypersensitivity (DTH), by day 9 of infection. Thereafter, responses of these mouse strains diverge. Whereas DTH persists in DBA/2 mice, BALB/c mice become anergic. Dr. Mitchell has found that these differences extend into the types of cytokines produced during active disease. DBA/2 mice produce IFN- earlier in infection and at higher levels than BALB/c mice; conversely, BALB/c mice produce IL-4 earlier and at higher levels than DBA/2 mice. The importance of these cytokines, as determinants of host response to disease, was established by showing that protection is potentiated in BALB/c mice by either treatment with recombinant IFN- , recombinant IL-12, or by in vivo depletion of IL-4. Similarly, in resistant DBA/2 mice, in vivo depletion of IFN- exacerbated disease. Taken together, these data provide evidence that the preferential induction of these immunomodulatory cytokines may have profound effects on the course of disease. Two specific aims are proposed. The first aim will be to delineate the kinetics and magnitude of cytokine expression in BALB/c and DBA/2 mice challenged by the pulmonary and systemic routes. Cytokine-specific mRNA will be measured by reverse-transcription polymerase chain reaction (RT- PCR) of RNA isolated from tissues and from unfractionated cells and purified cell populations from lungs, spleen, liver, and lymph nodes. Production of cytokine protein will be quantified by enzyme-linked immunosorbent assays (ELISA) of homogenates prepared from these tissues. The second aim will be to assess the biological relevance of individual cytokines on the host response by in vivo depletion of cytokines with neutralizing anti-cytokine antibodies. The effects of these interventions will be assessed by determining fungal load and cytokine modulation at the molecular and protein levels in tissues. Dr. Magee argues that completion of these aims will delineate new and fundamentally important aspects of mechanisms of host defenses in coccidioidomycosis and have significant implications for immunotherapeutic intervention in this disease.

描述（改编自申请人摘要）：宿主针对 C. 免疫炎是由特异性致敏的 T 细胞介导的。 在鼠标中 模型，具有遗传易感性（BALB/c）和遗传抗性 (DBA/2) 小鼠产生细胞介导的免疫 (CMI)，通过延迟- 型超敏反应 (DTH)，感染第 9 天。 此后， 这些小鼠品系的反应存在差异。 而 DTH 在 DBA/2 中仍然存在 小鼠、BALB/c 小鼠变得无反应。 米切尔博士发现这些 差异延伸到活动期间产生的细胞因子类型 疾病。 DBA/2 小鼠在感染早期和更高水平产生 IFN- 水平高于 BALB/c 小鼠；相反，BALB/c 小鼠更早产生 IL-4 且水平高于 DBA/2 小鼠。 这些的重要性 细胞因子作为宿主对疾病反应的决定因素被确立 通过证明 BALB/c 小鼠的保护作用通过以下任一方式得到增强 用重组 IFN-、重组 IL-12 或体内治疗 IL-4 耗竭。 同样，在耐药 DBA/2 小鼠中，体内耗竭 IFN-加剧疾病。 综合起来，这些数据提供了 有证据表明优先诱导这些免疫调节剂 细胞因子可能对疾病进程产生深远的影响。 提出了两个具体目标。 第一个目标是描绘 BALB/c 和 DBA/2 小鼠细胞因子表达的动力学和强度 受到肺部和全身途径的挑战。 细胞因子特异性 mRNA 将通过逆转录聚合酶链式反应（RT- PCR）从组织和未分级细胞中分离的 RNA 以及 从肺、脾、肝和淋巴结中纯化细胞群。 细胞因子蛋白的产生将通过酶联定量 对这些组织制备的匀浆进行免疫吸附测定 (ELISA)。 第二个目标是评估个体的生物学相关性 通过体内消耗细胞因子来影响细胞因子对宿主反应的影响 中和抗细胞因子抗体。 这些的影响 将通过确定真菌负荷和细胞因子来评估干预措施 在组织中的分子和蛋白质水平的调节。 马吉博士认为，这些目标的完成将描绘出新的和 宿主防御机制的根本重要方面 球孢子菌病并具有重大影响 对这种疾病进行免疫治疗干预。