Characteristics of Candida albicans adherence to mouse tissues
Characteristics of Candida albicans adherence to mouse tissues
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DOI：
10.1128/iai.58.6.1902-1908.1990
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发表时间：
1990-06
影响因子：
3.1
通讯作者：
J. Cutler;D. L. Brawner;Kevin C. Hazen;M. Jutila
中科院分区：
文献类型：
--
作者：
J. Cutler;D. L. Brawner;Kevin C. Hazen;M. Jutila
最初描述用于确定淋巴细胞归巢受体的离体结合试验适用于研究未固定组织切片中的白色念珠菌-宿主细胞相互作用。处死BALB/cByJ小鼠，取出各种器官，在干冰上快速冷冻并切片。C.将白色念珠菌酵母细胞悬浮在补充有5%新生小牛血清的Dulbecco改良Eagle培养基中，浓度为1.5 x 10（8）个细胞/ml，并将100微升悬浮液添加到组织切片中，在4摄氏度或22至24摄氏度下旋转15分钟。然后将切片固定在戊二醛中，清洗并检查。静止期酵母细胞比对数期细胞粘附得更好，并且在4 ℃和22至24 ℃下的粘附特性相似。从9株C.白色念珠菌显示相似的组织特异性。与淋巴结组织的粘连仅限于包膜下间隙和小梁窦。在脾脏中，酵母细胞与边缘区结合。在这两种组织中，酵母细胞与组织巨噬细胞的协会建议与巨噬细胞特异性单克隆抗体和荧光或免疫过氧化物酶染色技术的结果。C.白色念珠菌粘附于肾曲小管、肾小球和小动脉的图尼卡中膜。在小鼠实验性真菌血症中，C.除了在体内试验中没有观察到与肾小动脉结合之外，白色念珠菌酵母细胞与离体测定中相同的组织部位结合。一株酿酒酵母菌表现出与C.白色念珠菌，这表明组织粘附不足以产生毒力。附着机制尚未确定，但C。白色念珠菌的附着能力在数量上不同，二价阳离子的螯合剂抑制了结合。
An ex vivo binding assay originally described for determining lymphocyte homing receptors was adapted for studying Candida albicans-host cell interactions in unfixed tissue sections. BALB/cByJ mice were sacrificed, and various organs were removed, rapidly frozen on dry ice, and sectioned. C. albicans yeast cells were suspended to 1.5 x 10(8) cells per ml in Dulbecco modified Eagle medium supplemented with 5% newborn calf serum, and 100 microliters of the suspension was added to tissue sections for 15 min with rotation at 4 degrees C or at 22 to 24 degrees C. The sections were then fixed in glutaraldehyde, washed, and examined. Stationary-phase yeast cells adhered better than log-phase cells, and adherence characteristics were similar at 4 degrees C and 22 to 24 degrees C. Yeast cells from nine strains of C. albicans showed similar tissue specificity. Adherence to lymph node tissue was confined to subcapsular spaces and trabecular sinuses. In the spleen, yeast cells bound to the marginal zones. In both tissues, an association of yeast cells with tissue macrophages was suggested by results with macrophage-specific monoclonal antibodies and fluorescent or immunoperoxidase staining techniques. C. albicans adhered to convoluted tubules, glomeruli, and the tunica media of arterioles in the kidney. During experimentally induced fungemia in mice, C. albicans yeast cells associated with the same tissue sites as in the ex vivo assay, except that binding to renal arterioles was not seen in the in vivo test. A strain of Saccharomyces cerevisiae showed some adherence patterns in common with C. albicans, which indicates that tissue adherence is not sufficient for virulence. Mechanisms of attachment were not determined, but strains of C. albicans varied quantitatively in their ability to attach, and binding was inhibited by chelators of divalent cations.