Vadivelu Jamuna, Vellasamy Kumutha Malar, Thimma Jaikumar, Mariappan Vanitha, Kang Wen-Tyng, Choh Leang-Chung, Shankar Esaki M, Wong Kum Thong
Department of Medical Microbiology, Faculty of Medicine, University of Malaya Kuala Lumpur, Malaysia.
Division of Infection Biology, Department of Life Sciences, Central University of Tamil Nadu (CUTN), Thiruvarur, India.
PLoS Negl Trop Dis. 2017 Jan 3;11(1):e0005241. doi: 10.1371/journal.pntd.0005241. eCollection 2017 Jan.
During infection, successful bacterial clearance is achieved via the host immune system acting in conjunction with appropriate antibiotic therapy. However, it still remains a tip of the iceberg as to where persistent pathogens namely, Burkholderia pseudomallei (B. pseudomallei) reside/hide to escape from host immune sensors and antimicrobial pressure.
We used transmission electron microscopy (TEM) to investigate post-mortem tissue sections of patients with clinical melioidosis to identify the localisation of a recently identified gut microbiome, B. pseudomallei within host cells. The intranuclear presence of B. pseudomallei was confirmed using transmission electron microscopy (TEM) of experimentally infected guinea pig spleen tissues and Live Z-stack, and ImageJ analysis of fluorescence microscopy analysis of in vitro infection of A549 human lung epithelial cells.
TEM investigations revealed intranuclear localization of B. pseudomallei in cells of infected human lung and guinea pig spleen tissues. We also found that B. pseudomallei induced actin polymerization following infection of A549 human lung epithelial cells. Infected A549 lung epithelial cells using 3D-Laser scanning confocal microscopy (LSCM) and immunofluorescence microscopy confirmed the intranuclear localization of B. pseudomallei.
B. pseudomallei was found within the nuclear compartment of host cells. The nucleus may play a role as an occult or transient niche for persistence of intracellular pathogens, potentially leading to recurrrent episodes or recrudescence of infection.
在感染过程中,通过宿主免疫系统与适当的抗生素治疗协同作用可成功清除细菌。然而,对于持久性病原体——类鼻疽伯克霍尔德菌(B. pseudomallei)藏身何处以逃避宿主免疫传感器和抗菌压力,这仍然只是冰山一角。
我们使用透射电子显微镜(TEM)研究临床类鼻疽病患者的尸检组织切片,以确定最近鉴定出的肠道微生物群B. pseudomallei在宿主细胞内的定位。通过对实验感染的豚鼠脾脏组织进行透射电子显微镜(TEM)、活细胞Z轴扫描以及对A549人肺上皮细胞体外感染进行荧光显微镜分析并用ImageJ软件分析,证实了B. pseudomallei存在于细胞核内。
透射电子显微镜研究揭示了B. pseudomallei在受感染的人肺和豚鼠脾脏组织细胞的细胞核内定位。我们还发现,B. pseudomallei感染A549人肺上皮细胞后可诱导肌动蛋白聚合。使用3D激光扫描共聚焦显微镜(LSCM)和免疫荧光显微镜对感染的A549肺上皮细胞进行观察,证实了B. pseudomallei的细胞核内定位。
在宿主细胞的核区发现了B. pseudomallei。细胞核可能作为细胞内病原体持续存在的隐匿或短暂生态位,可能导致感染复发或病情复发。