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Dynamic expression of cell-surface antigens probed with Candida albicans-specific monoclonal antibodies

机译:用念珠菌特异性单克隆抗体探测细胞表面抗原的动态表达

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IgG hybridomas were produced which preferentially reacted with cell-surface antigens of either yeast cells or hyphae of Candida albicans. Four mAbs were used in an immunostaining procedure to follow the expression dynamics of these antigens in media supplemented with glucose or galactose. Yeast cell growth was analysed during the lag phase, the early- and late-exponential phases and the stationary phase, and mycelium formation was analysed between 0.5 and 24 h induction at 37 0C. It appears that yeast cell-surface antigens 5C11 and 2E11 are expressed throughout all phases of yeast cell growth as well as on young hyphae after up to 1 h induction. Longer hyphae only faintly react with these two mAbs as they switch to hyphal cell-surface antigens 2G8 and 4E1 after 3 h induction. The reactivity to mAbs 2G8 and 4E1 was induced after a 3 h temperature shift and was confined to the terminal third of growing mycelia. Growth and hyphae induction in galactose prolonged the reactivity of young hyphae with the two anti-yeast-cell mAbs, whereas the expression of surface antigens 2G8 and 2E11 appeared delayed and desynchronized on hyphae. Whereas a similar reactivity was found with ten ATCC strains of C. albicans, four clinical isolates had a unique pattern of reactivity. Immunoblot analyses of DTT extracts of cell-surface constituents indicated that the antigens were proteinaceous in nature and showed that yeast-cell antigens 5C11 and 2E11 are detected in four bands between 68 and 104 kDa, whereas mycelial antigens 4E1 and 2G8 are detected in 117 kDa and 104 kDa bands found in mycelial but not in yeast-cell extracts. Present data support the concept of a dynamic balance in the expression of phase-specific antigens in C. albicans.
机译:产生IgG杂交瘤,其优先与酵母细胞或念珠菌蛋白染蛋白的细胞表面抗原反应。在免疫染色过程中使用四种mAb以遵循补充有葡萄糖或半乳糖的培养基中这些抗原的表达动态。在滞后期期间分析酵母细胞生长,在37 0℃下分析早期和晚期阶段和固定相,和菌丝形成0.5至24小时诱导。似乎酵母细胞表面抗原5C11和2E11在酵母细胞生长的所有阶段表达,以及在高达1小时内的年轻菌丝上表达。更长的菌丝只有在3小时内切换到亚腿细胞表面抗原2G8和4E1时,仍然与这两种MAb进行微弱地反应。在3小时温度偏移后诱导对MAb 2G8和4E1的反应性,并限制在生长菌丝体的末端。半乳糖的生长和菌丝诱导延长了幼苗与两种抗酵母细胞mAb的反应性,而表面抗原2G8和2E11的表达出现在菌丝上延迟和去异步。虽然发现了具有10个ATCC的C. albicans的ATCC菌株的类似反应性,但是四个临床分离物具有独特的反应性模式。细胞表面成分的DTT提取物的免疫印迹分析表明,抗原本质上是蛋白质,并且显示酵母 - 细胞抗原5c11和2e11在68和104kda之间的四个条带中检测到,而在117kda中检测到菌丝体抗原4e1和2g8在菌丝体中发现104个KDA带,但不在酵母细胞提取物中。目前数据支持在C. albicans中表达相对相抗原的动态平衡的概念。

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