CANDIDA ADHERENCE MYCOLOGY RESEARCH UNIT

真菌学研究单位（MRU）的建议集中在一个全面的努力，以确定和评估候选抗原的能力，以产生保护性免疫对血源性播散性念珠菌病的能力。这一努力背后的驱动力是念珠菌感染的高频率，未来DNA疫苗策略的吸引力，以及对将抗真菌耐药性发展降到最低的治疗方法的需求。项目1建立在当前资助期发现的基础上，即抗体对白色念珠菌磷酸甘露聚糖复合物的某些表位的反应增强了对实验性播散性念珠菌病的抵抗力。拟议期间的方法包括：i)使用稳定的脂质体构建物来改进脂质体疫苗配方，ii)生产关键甘露聚糖表位的蛋白质偶联物，以及iii)基于“保护性”甘露聚糖表位的肽模位构建DNA疫苗。项目2是基于“保护性”甘露聚糖表位的ASL（凝集素半模位）的中心假设。项目2基于一个中心假设，即念珠菌的ASL（凝集素样序列）基因家族包含编码念珠菌对多种宿主成分的显性粘附的基因。ALS基因家族的基因产物将被评估为主动免疫和被动免疫的潜在疫苗靶点。主动免疫将使用DNA疫苗方法完成，并可与项目1中鉴定的磷酸甘露聚糖抗原结合使用，以优化免疫反应。项目3将利用从人血浆中获得的大量抗甘露聚糖抗体的亲和纯化能力，直接检测这些抗体的生物活性，以及对人抗甘露聚糖抗体优良表位特异性保护的贡献。项目4利用高度创新的分子生物学策略来鉴定可能成为有吸引力的疫苗靶点的尚未发现的细胞表面蛋白质。在感染过程中表达的基因将通过筛选随机融合基因来确定。将功能序列与白色念珠菌基因组序列进行比较，可以鉴定可能与抗体相互作用的分泌蛋白、细胞壁蛋白和跨膜蛋白。这些基因产物可以单独使用，也可以与其他目标免疫原联合使用，以开发有效的疫苗。这项技术很好地将多种候选免疫原结合到DNA疫苗中。

This proposal for a Mycology Research Unit (MRU) is focused on a comprehensive effort to identify and evaluate the ability the ability of candidate antigens to produce protective immunity against hematogenously disseminated candidiasis. The driving forces behind this effort are the high frequency of candidal infections, the attractiveness of a future DNA vaccine strategy, and the need for treatment approaches that will minimize the development of antifungal resistance. Project 1 builds on the discovery during the current grant period that antibody response to certain epitopes of the phosphomannan complex of C. albicans enhances resistance to experimental disseminated candidiasis. Approaches for the proposed period include: i) use of stabilized liposomal constructs to improve the liposomal vaccine formulations, ii) production of protein conjugates of the critical mannan epitopes, and iii) construction of a DNA vaccine based on peptide mimotopes of the "protective" mannan epitopes. Project 2 is based on the central hypothesis that the ASL (agglutinin mimotopes of the "protective" mannan epitopes. Project 2 is based on the central hypothesis that the ASL (agglutinin like sequence) gene family of Candida contains genes that encode dominant adhesions of Candida for a variety of host constituents. Gene products of the ALS gene family will be evaluated as potential vaccine targets for both active and passive immunization. The active immunization will be accomplished using DNA vaccine approaches and may be used in combination with phosphomannan antigens identified in project 1 to optimize an immune response. Project 3 will utilize the ability of affinity-purify large amounts of anti-mannan antibodies from human plasma to directly test the biological activities of these antibodies and the contribution to protection of the fine epitope specificity of human anti-mannan antibody. Project 4 utilizes highly innovative molecular biology strategies to identify yet undiscovered cell surface proteins that may be attractive vaccine targets. Genes that are expressed during infection will be identified by screening of random fusion genes. Comparison of functional sequences to the C. albicans genomic sequence will permit identification of likely secreted, cell wall, and transmembrane proteins, available for interaction with antibody. These gene products can then be used either alone or in combination with other target immunogens to develop effective vaccines. This technology lends itself well to the incorporation of multiple candidate immunogens into DNA vaccines.