Candida in vivo expressed protein as a mannan carrier

念珠菌病在免疫功能低下的患者中是个问题，表现为从危及生命的粘膜皮肤到危及生命的血源性传播感染的疾病。念珠菌病的主要病原体是白色念珠菌，尽管有几种念珠菌很重要，但我们的研究将集中在念珠菌上。白色念珠菌以前，我们提供了保护性疫苗的概念证明，但我们提出的工作将导致疫苗配方的基本改进。最初的疫苗利用牛血清白蛋白（BSA）作为蛋白载体，与粗真菌甘露聚糖组分偶联，但计划的改进包括用C.在疾病发病过程中表达的白念珠菌表面蛋白，以及合成的β-连接的寡甘露糖苷代替粗甘露聚糖。令人兴奋的方面， 我们的工作是与其他MRU项目的相互作用将使我们能够鉴定已知在人类念珠菌病发病过程中表达的真菌细胞壁表面蛋白，因为到目前为止，我们所有的工作都局限于这种人类疾病的小鼠模型。这项工作是由两个假设驱动的：用适当的体内（人）表达的C取代BSA。白色念珠菌细胞壁表面蛋白将导致针对蛋白质载体的保护性应答;并且，用合成的β-连接的甘露糖苷替代粗甘露聚糖部分将有利于保护性抗体应答的产生。这两个假设将通过以下具体目标进行检验。1.利用IVIAT和蛋白质微阵列来识别 候选真菌载体蛋白。2.通过利用细胞壁制备物的2-D凝胶电泳来确定没有广泛N-糖基化的蛋白质，从而细化目标1中定义的候选蛋白质的数量。3.疫苗将潜在的载体蛋白偶联到甘露聚糖提取物上，并在小鼠中测试偶联物的疫苗效力。4.用合成β-甘露糖苷代替甘露聚糖提取物。这些目标的实现应导致疫苗的发展，防止粘膜和传播形式的念珠菌病。

Candidiasis is problematic amongst immunocompromised patients, manifesting as disease ranging from life-compromising mucocutaneous to life-threatening hematogenously disseminated infections. The primary agent of candidiasis is Candida albicans and, although several species of Candida are important, our studies will focus on C. albicans. Previously we provided proof of concept for a protective vaccine, but our proposed work will result in essential refinement of the vaccine formulation. The original vaccine utilized bovine serum albumin (BSA) as a protein carrier coupled to a crude fungal mannan fraction, but the planned improvements include replacement of the BSA with a C. albicans surface protein expressed during pathogenesis of disease, and synthetic beta-linked oligomannosides instead of crude mannan. An exciting aspect of the work is that interaction with the other MRU projects will allow us to identify fungal cell wall surface proteins that are known to be expressed during pathogenesis of candidiasis in humans, as until now all of our work has been restricted to mouse models of this human disease. The work is driven by two hypotheses: replacement of the BSA with an appropriate in vivo (human) expressed C. albicans cell wall surface protein will lead to protective responses against the protein carrier; and, replacement of the crude mannan portion with synthetic beta-linked mannosides will favor production of protective antibody responses. The two hypotheses will be tested by the following specific aims. 1. Utilize IVIAT and protein microarrays to identify candidate fungal carrier proteins. 2. Refine the number of candidate proteins defined in aim 1 by utilizing 2-D gel electrophoresis of cell wall preparations to identify proteins without extensive N-glycosylation. 3. Covalently couple potential carrier proteins to mannan extracts and test the conjugates in mice for vaccine efficacy. 4. Replace the mannan extract with synthetic beta-mannosides. Fulfillment of these aims should lead to development of a vaccine that prevents both mucosal and disseminated forms of candidiasis.