摘要
目的:研究鼠疫耶尔森菌(鼠疫菌)在巨噬细胞内连续传代诱导后的表型变化情况。方法:布氏田鼠型鼠疫菌201株在小鼠巨噬细胞Raw264.7中连续传代50次后,获得诱导株(201-MI)。通过巨噬细胞内生存能力、生长曲线测定、生物膜形成能力、酸生存能力、过氧化氢生存能力、对Raw264.7及HeLa细胞的毒力以及小鼠攻毒等实验,比较野生株201和诱导株201-MI的特定表型差异。并利用全基因组测序初步分析了201株与201-MI株的遗传差异。结果:与野生型201株相比,201-MI株的巨噬细胞生存能力显著增强,在铁(Fe)缺乏培养基中生长较快,酸生存及过氧化氢耐受能力增强,生物膜形成能力减弱,对Raw264.7和HeLa细胞的损伤程度降低,但对小鼠毒力减弱(皮下攻毒与腹腔攻毒)。比较基因组分析未发现201-MI株存在基因片段的缺失,但存在一些小的插入/缺失(Indel)和单核苷酸突变(SNP)。结论:巨噬细胞连续传代诱导后,鼠疫菌出现了一些表型变化,可能反映了其巨噬细胞压力下的适应性进化。对诱导菌株进行后续多组学研究将有助于了解这些变化的遗传机制,以及鼠疫菌-巨噬细胞相互作用的生物学过程。
Objective To investigate the changes in adaptive phenotypes of Yersinia pestis(Yp)during successive passages in macrophages.Methods A Yp strain of 201-MI was induced by 50 successive passages of Yp 201 strain in Raw264.7 cells.Phenotypic characteristics of 201 and 201-MI strains were compared by analyzing their survival rates in macrophages,growth curves,biofilm formation abilities,acid and hydrogen peroxide-stress tolerance,and virulence to mammal cells(Raw264.7 and HeLa cells)and mice.Results Comparing with 201 strain,201-MI strain showed various phenotypic changes,including higher survival rate in Raw264.7 cells,faster growth in iron-deficient medium,higher tolerance to acid and hydrogen peroxide,decreased biofilm formation ability,and less damages to Raw264.7 and HeLa cells.More-over,201-MI strain showed decreased virulence to mice in both subcutaneous and intraperitoneal challenges.Preliminary comparative genomics analysis revealed some indel and nonsense mutations in 201-MI strain,which might account for its phenotype changes.Conclusions After successive passages in macrophages,Yp showed some phenotypic changes,which might reflect its adaptive evolution under the pressure of macrophages.Detailed multi-omics analysis would be of great help to understand the underlying genetic mechanisms of these changes,and the related Yp-macrophage interaction processes as well.
作者
陈欣
宋凯
武雅蓉
肖丽婷
金钧妍
杜奕溥
崔玉军
余莉
宋亚军
Chen Xin;Song Kai;Wu Yarong;Xiao Liting;Jin Junyan;Du Yipu;Cui Yujun;Yu Li;Song Yajun(School of Basic Medical Sciences,Anhui Medical University,Hefei 230032,China;Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences,Beijing 100071,China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2022年第4期251-257,共7页
Chinese Journal of Microbiology and Immunology
关键词
鼠疫菌
巨噬细胞
适应性变化
毒力
Yersinia pestis
Macrophages
Adaptive changes
Virulence
