粒细胞-巨噬细胞集落刺激因子缺陷对小鼠寡育外瓶霉感染病程及免疫应答的影响  

Impact of GM-CSF deficiency on the disease course and immune response in mice infected with Exophiala oligosperma

作　　者：董琪 陆捷洁 吴伟伟 张瑞珺 DONG Qi;LU Jiejie;WU Weiwei;ZHANG Ruijun(Third Hospital of Shanxi Medical University,Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Tongji Shanxi Hospital,Taiyuan,Shanxi 030032,China;Department of Dermatology,Affiliated Dermatology Hospital of Hainan Medical University,Department of Dermatology,the Fifth People's Hospital of Hainan Province,Haikou,Hainan 570206,China)

机构地区：[1]山西医科大学第三医院,山西白求恩医院(山西医学科学院),同济山西医院,山西太原030032 [2]海南医科大学附属皮肤病医院皮肤科,海南省第五人民医院皮肤科,海南海口570206

出　　处：《中国热带医学》2025年第1期28-35,共8页China Tropical Medicine

基　　金：国家自然科学基金项目(No.82002125,No.82160393);山西省基础研究计划项目(No.202303021221196);海南省自然科学基金项目(No.822CXTD536)。

摘　　要：目的探讨粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony-stimulating factor,GM-CSF)缺乏在暗色真菌寡育外瓶霉(Exophiala oligosperma)感染发病机制中的作用,以期为暗色真菌感染的治疗提供新的思路和依据。方法选取C57BL/6野生型(wild type,WT)小鼠和Csf2基因敲除(knock out,KO)小鼠(C57BL/6遗传背景),利用从胱天蛋白酶募集域蛋白9(caspase recruitment domain-containing protein 9,CARD9)基因缺陷患者中分离的暗色真菌寡育外瓶霉,通过建立小鼠皮下感染模型,模拟人类感染途径。对感染小鼠的自然病程持续观测6周,并于适当节点取皮损组织进行病理学分析和免疫反应监测。结果WT小鼠和Csf2 KO小鼠在病程中均呈现自愈倾向,病原体自发清除,足部组织外观逐渐恢复正常,在观察终点生存率为100%。与WT小鼠相比,Csf2 KO小鼠的足部肿胀率在感染1、2周时较低,差异有统计学意义(t=4.674,t=5.961,P<0.01);Csf2 KO小鼠的真菌清除速度较慢,第4周时仍能在皮损组织中检测到真菌菌落,并且组织病理镜下观察可见过碘酸雪夫(periodic acid-schiff,PAS)染色阳性的真菌孢子结构;Csf2 KO小鼠早期(1~2周)巨噬细胞浸润的差异无统计学意义(P>0.05),而中性粒细胞浸润在感染2周时显著减少(t=3.287,P<0.01);在足部皮损匀浆液中,感染1周时,Csf2 KO小鼠IL-6和IL-1β水平降低(t=4.686,t=4.102,P<0.05)。结论本研究发现GM-CSF缺乏可延缓病原真菌的清除,延长病程,影响感染早期炎症因子的产生以及中性粒细胞的浸润。Objective To investigate the role of granulocyte-macrophage colony-stimulating factor(GM-CSF)deficiency in the pathogenesis of Exophiala oligosperma(E.oligosperma)infection,a dematiaceous fungus,aiming to provide new insights and evidence for the treatment of dematiaceous fungal infections.Methods C57BL/6 wild-type(WT)mice and Csf2 gene knockout(KO)mice(C57BL/6 background)were selected.Using E.oligosperma isolated from patients with caspase recruitment domain-containing protein 9(CARD9)gene deficiency,a murine subcutaneous infection model was established to simulate the human infection route.The natural progression of infection in the mice was observed for six weeks,with skin lesion tissues collected at appropriate time points for pathological analysis and monitoring immune responses.Results Both WT and Csf2 KO mice exhibited spontaneous pathogen clearance and gradual recovery of foot tissue appearance during the progression of infection,with a 100%survival rate at the end of observation.Compared to WT mice,Csf2 KO mice showed reduced footpad swelling at 1 and 2 weeks post-infection(t=4.674,t=5.961,P<0.01).Fungal clearance in Csf2 KO mice was delayed,with fungal colonies still detectable in lesion tissues at week 4,and Periodic Acid-Schiff(PAS)-positive fungal spores observed in histopathological sections.There was no significant difference found in macrophage infiltration between WT and Csf2 KO mice during the early stages of infection(1-2 weeks)(P>0.05),while neutrophil infiltration was significantly reduced in Csf2 KO mice at week 2(t=3.287,P<0.01).In addition,Csf2 KO mice exhibited lower levels of IL-6 and IL-1βin foot lesion homogenates at week 1(t=4.686,t=4.102,P<0.05).Conclusions This study demonstrated that GM-CSF deficiency delays pathogenic fungi clearance,prolongs the disease course and affects early inflammatory cytokine production as well as neutrophil infiltration during the early stages of fungal infections.

关 键 词：粒细胞-巨噬细胞集落刺激因子 暗色真菌 寡育外瓶霉 固有免疫反应 中性粒细胞 

分 类 号：R519[医药卫生—内科学]