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一个问题，表现为从危及生命的粘膜皮肤到危及生命的血液播散性感染。念珠菌病的主要病原体是白色念珠菌，虽然有几种念珠菌很重要，但我们的研究将集中在白色念珠菌上。以前，我们提供了保护性疫苗的概念证明，但我们提议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）作为蛋白质载体偶联于粗真菌甘露聚糖部分，但计划中的改进包括用在疾病发病过程中表达的白色念珠菌表面蛋白替代BSA，以及合成β -连接寡糖甘露聚糖代替粗甘露聚糖。一个令人兴奋的方面

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.