Weinberg P B, Becker S, Granger D L, Koren H S
Center for Environmental Medicine, University of North Carolina at Chapel Hill.
Am Rev Respir Dis. 1987 Nov;136(5):1242-7. doi: 10.1164/ajrccm/136.5.1242.
Macrophage cytotoxicity for Cryptococcus neoformans was investigated by culturing human alveolar macrophage (AM) with a thin-capsuled clone of C. neoformans in a polypropylene culture tube assay system. Yeast replication was quantitated by electronic particle counting after detergent lysis of AM and viability by quantitative plate counts. Under appropriate conditions, fungal replication was inhibited in the presence of human AM. This effect persisted over the 48-h time course that was evaluated. During this period, organisms in medium alone proliferated rapidly, doubling their number every 4 h. Human AM did not require endotoxin, fetal calf serum, or specific rabbit anticryptococcal antibody for fungistasis. Under these conditions, microscopic evaluation of a cytocentrifuge preparation of AM-yeast cocultures, stained by a modified Giemsa technique, revealed all the fungi to be extracellular. In the presence of 10% fresh human serum, AM phagocytized C. neoformans and exhibited fungicidal activity. Tumor necrosis factor did not affect the replication rate of the yeast. These findings suggest that there may be at least 2 mechanisms by which human AM protect against C. neoformans. One is serum-independent and extracellular and results in fungistasis, and the other is dependent on a serum factor and leads to intracellular inhibition of growth and possibly killing of the organism.
通过在聚丙烯培养管检测系统中，将人肺泡巨噬细胞（AM）与新型隐球菌的薄荚膜克隆进行共培养，研究了巨噬细胞对新型隐球菌的细胞毒性。在对AM进行去污剂裂解后，通过电子颗粒计数对酵母复制进行定量，并通过定量平板计数对活力进行测定。在适当条件下，人AM存在时真菌复制受到抑制。在评估的48小时时间进程中，这种效应持续存在。在此期间，仅在培养基中的生物体迅速增殖，每4小时数量翻倍。人AM抑制真菌生长不需要内毒素、胎牛血清或特异性兔抗隐球菌抗体。在这些条件下，用改良吉姆萨技术染色的AM-酵母共培养物的细胞离心涂片的显微镜评估显示，所有真菌均位于细胞外。在10%新鲜人血清存在的情况下，AM吞噬新型隐球菌并表现出杀真菌活性。肿瘤坏死因子不影响酵母的复制速率。这些发现表明，人AM抵御新型隐球菌可能至少有两种机制。一种是不依赖血清且在细胞外，导致真菌生长抑制，另一种依赖血清因子，导致细胞内生长抑制并可能杀死生物体。
