CANDIDA ADHERENCE MYCOLOGY RESEARCH UNIT

这项关于真菌学研究单位(MRU)的提议侧重于一项综合努力，以确定和评估候选抗原产生针对血源性播散性念珠菌病的保护性免疫的能力。这一努力背后的驱动力是念珠菌感染的高频率，未来DNA疫苗战略的吸引力，以及将抗真菌耐药性的发展降至最低的治疗方法的需要。项目1的基础是在目前的赠款期间发现，对白色念珠菌的磷甘露聚糖复合体的某些表位的抗体反应增强了对实验性播散性念珠菌病的抵抗力。建议的方法包括：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结合物，以及iii)基于保护性甘露聚糖表位的多肽模拟表位构建DNA疫苗。项目2基于的中心假设是，甘露聚糖的“保护性”表位的凝集素模拟表位。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各种宿主成分的显性黏附的基因。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中确定的磷甘露聚糖抗原结合使用，以优化免疫反应。项目三将利用从人血浆中亲和纯化大量抗甘露聚糖抗体的能力，直接测试这些抗体的生物学活性，并为保护人抗甘露聚糖抗体良好的表位特异性做出贡献。项目4利用高度创新的分子生物学策略来确定尚未发现的细胞表面蛋白，这些蛋白可能是有吸引力的疫苗靶标。在感染期间表达的基因将通过随机融合基因的筛选来识别。将功能序列与白念珠菌基因组序列进行比较，可以鉴定出可能的分泌型、细胞壁和跨膜蛋白，可与抗体相互作用。然后，这些基因产品可以单独使用，也可以与其他目标免疫原结合使用，以开发有效的疫苗。这项技术很好地将多种候选免疫原整合到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.