PHOSPHOMANNOPROTEIN ADHESIN AS A VACCINE CANDIDATE

血源性播散性念珠菌病的发病机制似乎是 涉及白念珠菌酵母细胞与白念珠菌之间的黏附事件 特定的宿主组织。我们假设宿主抗体是针对 念珠菌粘附素改变了发病机制，可能有助于宿主的生存。在……里面 PI的实验室，假丝酵母菌粘附素已经被分离出来 小鼠脾边缘带巨噬细胞与酵母细胞的特异性黏附。 这些粘附素是磷脂蛋白(PMP)复合体的一部分 念珠菌细胞表面。增溶粘附素制成的疫苗 脂质体包裹后在小鼠体内引起抗体反应 粘附素。接种疫苗的动物增加了对 播散性念珠菌病，他们的血清中和粘附素活性， 防止酵母细胞附着在脾上，并似乎转移 保护。抗PMP来源的单抗 粘附素在PI的实验室中可用。多克隆抗体的作用 而单抗与各种组织的黏附相互作用将是 通过依从性试验进行广泛评估。具体目标是：1. 确定保护性反应的最佳免疫方案 正常的小鼠。变量包括粘附素的组成、剂量、作用 佐剂、给药途径和助剂时间表。2.确定 接种疫苗在预防播散性念珠菌病中的作用 将在正常小鼠(BALB/c雌性)中评估免疫效果 和雄性小鼠和BALB/c杂交的外交系)。3.使用被动语态 转移实验以确定抗体是否对 豁免权。来自接种疫苗的动物的免疫血清，特异性的单抗 白念珠菌的粘附素和抗疏水蛋白的单抗将 测试了它们保护幼稚动物免受传播的能力 念珠菌病。免疫功能正常的小鼠，T细胞缺陷(nu/nu)， T和B细胞缺陷(SCID)和诱导性中性粒细胞减少症(MAb) RB6-8C5处理)将进行测试。4.对……展开调查 疫苗和单抗诱导的保护机制。体外实验 毛细管剪切黏附实验、内皮细胞黏附实验 将使用贴壁试验和活体内显微镜方法 检测免疫血清和保护性单抗对黏附的影响 白念珠菌对不同宿主细胞、组织和 糖蛋白。免疫血清和单抗对黏附的影响 补体调理细胞和未调理细胞的特性 接受检查。这一结果很可能导致新的预防和治疗方法 播散性念珠菌病的治疗策略。

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.