CARD9缺陷的暗色丝孢霉病患者中性粒细胞抗白念珠菌免疫功能的研究  

Functional studies of neutrophils from phaeohyphomycosis patients with CARD9 deficiencies against Candida albicans

作　　者：梁嫔[1,2] 王晓雯[1,2] 万喆[1,2] 李若瑜[1,2] 

机构地区：[1]北京大学第一医院皮肤科 [2]北京大学医学真菌研究中心皮肤病分子诊断北京市重点实验室,北京100034

出　　处：《中国真菌学杂志》2015年第1期1-5,共5页Chinese Journal of Mycology

基　　金：国家自然科学基金(81472890;81171510)

摘　　要：目的探讨CARD9缺陷的暗色丝孢霉病患者中性粒细胞抗白念珠菌的免疫功能。方法提取CARD9缺陷患者及正常人外周血中性粒细胞,并使其与白念珠菌进行孵育。采用菌落计数法及MTT法检测中性粒细胞对白念珠菌的杀伤,流式细胞术检测中性粒细胞活性氧的产生及吞噬能力,电镜观察吞噬溶酶体的超微结构,流式微球检测试剂盒检测培养上清中IL-8的分泌情况。结果与对照组比较,CARD9缺陷患者中性粒细胞对白念珠菌孢子的杀伤及IL-8的表达存在缺陷,菌丝杀伤能力、活性氧的产生、吞噬能力以及吞噬溶酶体的结构无明显异常,且血清调理后杀伤能力以及IL-8的表达得到恢复。结论 CARD9蛋白在中性粒细胞杀伤白念珠菌孢子免疫过程中发挥着关键作用,血清调理可恢复该缺陷。Caspase recruitment domain-containing protein 9 （ CARD9） is an adaptor molecule that is critical for NF-κB activa-tion,and forms a complex with B-cell lymphoma 10 （ BCL10） and mucosa-associated lymphoid tissue lymphoma translocation gene 1 （ MALT1） that mediates C-type lectin receptors （ CLRs）-triggered intracellular signaling during antifungal immunity. In the present study,we investigated the functions of polymorphonuclear neutrophils （ PMNs） from phaeohyphomycosis patients with CARD9 defi-ciencies against Candida albicans （C.albicans）.PMNs were isolated from patients and healthy blood donors and subsequently chal-lenged with C.albicans .By calculating colony-forming units and MTT assay,the killing of C.albicans by CARD9-deficient neutrophils was detected, and the ROS production and phagocytotic ability of CARD9-deficient neutrophils was assayed by Flow CytoMetry （FCM）.The phagolysosome ultrastructure and the secretion of IL-8 were analyzed separately by EM and CBA kit.As a result,we demonstrated that,compared with healthy donors,CARD9-deficient PMNs exhibited defects in C. albicans killing and IL-8 produc-tions,which can be rescued in the presence of serum;however,the CARD9-deficient PMNs exhibited normal reactive oxygen species generation,phagocytotic ability and phagolysosome ultrastructure.In conclusion,our results indicate that CARD9 is indispensable for C.albicans killing by PMNs,and serum-opsonization acts as a CARD9-independent way,which could be a promising immunotherapy in the future.

关 键 词：CANDIDA ALBICANS CARD9 中性粒细胞 ROS 吞噬 吞噬溶酶体 白细胞介素-8 

分 类 号：R379.4[医药卫生—病原生物学]