PHOSPHOMANNOPROTEIN ADHESIN AS A VACCINE CANDIDATE

血行播散性念珠菌病的发病机制似乎是 涉及白色念珠菌酵母细胞之间的粘附事件和 特定的宿主组织。 我们假设宿主抗体特异性 念珠菌粘附素改变发病机制并可能有助于宿主生存。 在 PI 的实验室已分离出念珠菌粘附素 特定的酵母细胞粘附于小鼠脾边缘区巨噬细胞。 这些粘附素是磷酸甘露糖蛋白 (PMP) 复合物的一部分 念珠菌细胞表面。 由溶解的粘附素制成的疫苗 封装在脂质体中可激发小鼠的抗体反应 粘附素。 接种疫苗的动物抵抗力增强 播散性念珠菌病，其血清中和粘附素活性， 防止酵母细胞附着到脾脏上，并且似乎转移 保护。 针对 PMP 衍生的单克隆抗体 (mAb) PI 实验室提供粘附素。 多克隆的影响 和单克隆抗体与各种组织的粘附相互作用将 通过依从性测定进行广泛评估。 具体目标是：1. 确定保护反应的最佳免疫方案 正常小鼠。 变量包括粘附素的组成、剂量、效果 佐剂、给药途径和加强方案。 2. 确定 疫苗接种在预防传播性念珠菌病中的效果。 将在正常小鼠（BALB/c 雌性小鼠）中评估免疫效果 以及雄性小鼠和 BALB/c 杂交的外面包种）。 3.使用被动语态 转移实验以确定抗体是否负责 免疫力。 来自接种疫苗的动物的免疫血清，针对该疾病的单克隆抗体 白色念珠菌的粘附素和针对疏水蛋白的单克隆抗体将 测试了它们保护幼稚动物免受传播的能力 念珠菌病。 免疫能力强的小鼠，T 细胞缺陷 (nu/nu)， T 细胞和 B 细胞缺陷 (SCID) 以及诱导性中性粒细胞减少症小鼠 (mAb RB6-8C5 处理）将进行测试。 4. 启动调查 疫苗和单克隆抗体诱导的保护机制。 离体 测定、毛细管剪切依赖性粘附测定、内皮细胞 将使用粘附试验和体内活体显微镜方法 确定免疫血清和保护性单克隆抗体对依从性的影响 白色念珠菌对各种宿主细胞、组织和细胞的特性 糖蛋白。 免疫血清和单克隆抗体对依从性的影响 补体调理细胞和非调理细胞的特征将 被检查。 研究结果很可能会带来新的预防和治疗方法 传播性念珠菌病的策略。

The pathogenesis of hematogenous disseminated candidiasis appears to involve adhesion events between yeast cells of Candida albicans and specific host tissues. We hypothesize that host antibodies specific for candidal adhesins alter the pathogenesis and may aid host survival. In the PI's laboratory, candidal adhesins have been isolated that cause specific yeast cell adherence to mouse splenic marginal zone macrophages. These adhesins are part of the phosphomannoprotein (PMP) complex on the candidal cell surface. Vaccines made of solubilized adhesins encapsulated in liposomes provoke antibody responses in mice against the adhesins. Vaccinated animals have increased resistance against disseminated candidiasis, their serum neutralizes adhesin activity, prevents yeast cell attachment to the spleen, and appears to transfer protection. Monoclonal antibodies (mAbs) against the PMP-derived adhesins are available in the PI's laboratory. The effects of polyclonal and mAbs on adherence interactions with various tissues will be extensively evaluated by adherence assays. The specific aims are to: 1. Determine the optimal immunization protocol for protective responses in normal mice. Variables include adhesin composition, dose, effect of adjuvants, routes of administration and booster schedule. 2. Determine the effect of the vaccination in prevention of disseminated candidiasis. Immunization effectiveness will be assessed in normal mice (BALB/c female and male mice and a BALB/c crossed outbread strain). 3. Use passive transfer experiments to determine if antibodies are responsible for immunity. Immune sera from vaccinated animals, mAbs specific for the adhesins of C. albicans, and mAbs against hydrophobic proteins will be tested for their ability to protect naive animals against disseminated candidiasis. Immunologically competent mice, T-cell deficient (nu/nu), T- and B- cell deficient (SCID), and mice with induced neutropenia (mAb RB6-8C5 treated) will be tested. 4. Initiate investigations on mechanisms of protection induced by the vaccine and mAbs. The ex vivo assay, the capillary tube shear-dependent adhesion assay, the endothelial adherence assay, and in vivo intravital microscopic method will be used to determine the effect of immune sera and protective mAbs on adherence characteristics of C. albicans to various host cells, tissues and glycoproteins. The effect of immune sera and mAbs on adherence characteristics of complement opsonized cells and unopsonized cells will be examined. The results may well lead to new preventive and therapeutic strategies for disseminated candidiasis.