Genetic Screening of Candida albicans Inactivation Mutants Identifies New Genes Involved in Macrophage-Fungal Cell Interactions.
Genetic Screening of Candida albicans Inactivation Mutants Identifies New Genes Involved in Macrophage-Fungal Cell Interactions.
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DOI：
10.3389/fmicb.2022.833655
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发表时间：
2022
影响因子：
5.2
通讯作者：
Whiteway, Malcolm
中科院分区：
文献类型：
--
作者：
Godoy, Pablo;Darlington, Peter John;Whiteway, Malcolm
关键词：
白色念珠菌是一种重要的人类真菌病原体，具有不同的形态，如酵母、假菌丝和菌丝，它们被天然免疫反应的吞噬细胞不同地识别。一旦白念珠菌细胞入侵宿主组织，巨噬细胞等免疫细胞就会被吸引到感染部位，并被激活以识别、吞噬和杀死病原体。我们通过高通量筛选白念珠菌GRACE文库来研究这种真菌细胞-巨噬细胞界面，以确定可以影响这种相互作用的基因，并修改吞噬动力学。与野生型(WT)菌株相比，我们发现那些具有组成假菌丝和菌丝表型的真菌菌株的吞噬速度普遍更快，而酵母形态锁定的菌株表现出被巨噬细胞识别和内化的减少和延迟。我们鉴定了一些GRACE菌株，它们的形态发育正常，但对培养的巨噬细胞表现出不同的识别和吞噬动力学，并鉴定了两个突变株，它们改变了与小鼠和人来源的巨噬细胞的相互作用。一个突变体失活了一个没有特征的白色念珠菌开放阅读框架，它是酿酒酵母OPY1的同源基因，另一个失活的CaKRE1。在4h的共培养过程中监测修改后的相互作用。在相互作用的早期，与WT细胞相比，opy1和kre1突变株对巨噬细胞的识别和吞噬速度都有所降低。在真菌芽管启动时，突变体和WT细胞的吞噬动力学都增加，但WT细胞在相互作用2小时内仍表现出较高的巨噬细胞内化能力。随后，在相互作用的2~4h之间，当大多数巨噬细胞含有被吞噬的真菌细胞时，opy1突变体的吞噬动力学增加，而kre1突变体的吞噬动力学进一步降低。表明真菌形态影响巨噬细胞与白念珠菌细胞的结合，OPY1和KRE1在真菌细胞与吞噬细胞的相互作用中发挥作用。
Candida albicans, an important fungal pathogen of humans, displays different morphologies, such as yeast, pseudo-hyphae and hyphae, which are recognized unequally by phagocytic cells of the innate immune response. Once C. albicans cells invade host tissues, immune cells such as macrophages are attracted to the site of infection and activated to recognize, engulf and kill the pathogen. We have investigated this fungal cell-macrophage interface by using high-throughput screening of the C. albicans GRACE library to identify genes that can influence this interaction and modify the kinetics of engulfment. Compared with the wild-type (WT) strain, we identified generally faster rates of engulfment for those fungal strains with constitutive pseudo-hyphal and hyphal phenotypes, whereas yeast-form-locked strains showed a reduced and delayed recognition and internalization by macrophages. We identified a number of GRACE strains that showed normal morphological development but exhibited different recognition and engulfment kinetics by cultured macrophages and characterized two mutants that modified interactions with the murine and human-derived macrophages. One mutant inactivated an uncharacterized C. albicans open reading frame that is the ortholog of S. cerevisiae OPY1, the other inactivated CaKRE1. The modified interaction was monitored during a 4 h co-culture. Early in the interaction, both opy1 and kre1 mutant strains showed reduced recognition and engulfment rates by macrophages when compared with WT cells. At fungal germ tube initiation, the engulfment kinetics increased for both mutants and WT cells, however the WT cells still showed a higher internalization by macrophages up to 2 h of interaction. Subsequently, between 2 and 4 h of the interaction, when most macrophages contain engulfed fungal cells, the engulfment kinetics increased for the opy1 mutant and further decreased for the kre1 mutant compared with Ca-WT. It appears that fungal morphology influences macrophage association with C. albicans cells and that both OPY1 and KRE1 play roles in the interaction of the fungal cells with phagocytes.
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影响因子：
11.8
作者：
Horn, David L.;Neofytos, Dionissios;Webster, Karen M.
通讯作者：
Webster, Karen M.
影响因子：
3.6
作者：
Hall RA
通讯作者：
Hall RA
影响因子：
16.6
作者：
Ding X;Kambara H;Guo R;Kanneganti A;Acosta-Zaldívar M;Li J;Liu F;Bei T;Qi W;Xie X;Han W;Liu N;Zhang C;Zhang X;Yu H;Zhao L;Ma F;Köhler JR;Luo HR
通讯作者：
Luo HR
影响因子：
2.8
作者：
Breinig, F;Schleinkofer, K;Schmitt, MJ
通讯作者：
Schmitt, MJ
影响因子：
6.7
作者：
Hopke A;Nicke N;Hidu EE;Degani G;Popolo L;Wheeler RT
通讯作者：
Wheeler RT