a Division of Pediatric Infectious Diseases , Vanderbilt University School of Medicine.
b Tennessee Valley Healthcare Systems , Department of Veterans Affairs.
Infect Dis (Lond). 2016;48(3):177-88. doi: 10.3109/23744235.2015.1100323. Epub 2015 Oct 22.
Bordetella pertussis colonizes the human respiratory mucosa. Most studies on B. pertussis adherence have relied on cultured mammalian cells that lack key features present in differentiated human airway cells or on animal models that are not natural hosts of B. pertussis. The objectives of this work were to evaluate B. pertussis infection in highly differentiated human airway cells in vitro and to show the role of B. pertussis fimbriae in cell adherence.
Primary human airway epithelial (PHAE) cells from human bronchi and a human bronchial epithelial (HBE) cell line were grown in vitro under air-liquid interface conditions.
PHAE and HBE cells infected with B. pertussis wild-type strain revealed bacterial adherence to the apical surface of cells, bacteria-induced cytoskeleton changes, and cell detachment. Mutations in the major fimbrial subunits Fim2/3 or in the minor fimbrial adhesin subunit FimD affected B. pertussis adherence to predominantly HBE cells. This cell model recapitulates the morphologic features of the human airway infected by B. pertussis and confirms the role of fimbriae in B. pertussis adherence. Furthermore, HBE cells show that fimbrial subunits, and specifically FimD adhesin, are critical in B. pertussis adherence to airway cells.
The relevance of this model to study host-parasite interaction in pertussis lies in the striking physiologic and morphologic similarity between the PHAE and HBE cells and the human airway ciliated and goblet cells in vivo. These cells can proliferate in vitro, differentiate, and express the same genetic profile as human respiratory cells in vivo.
百日咳博德特氏菌定植于人体呼吸道黏膜。大多数关于百日咳博德特氏菌黏附的研究依赖于培养的哺乳动物细胞,这些细胞缺乏存在于分化的人呼吸道细胞中的关键特征,或者依赖于不是百日咳博德特氏菌天然宿主的动物模型。本研究的目的是评估体外高度分化的人呼吸道细胞中的百日咳博德特氏菌感染,并证明百日咳博德特氏菌菌毛在细胞黏附中的作用。
体外采用空气-液体界面条件培养原代人呼吸道上皮(PHAE)细胞和人支气管上皮(HBE)细胞系。
感染百日咳博德特氏菌野生型菌株的 PHAE 和 HBE 细胞显示出细菌黏附在细胞的顶端表面、细菌诱导的细胞骨架变化和细胞脱落。主要菌毛亚基 Fim2/3 或次要菌毛黏附因子亚基 FimD 的突变影响了百日咳博德特氏菌对主要是 HBE 细胞的黏附。该细胞模型再现了被百日咳博德特氏菌感染的人呼吸道的形态特征,并证实了菌毛在百日咳博德特氏菌黏附中的作用。此外,HBE 细胞表明菌毛亚基,特别是 FimD 黏附因子,在百日咳博德特氏菌对气道细胞的黏附中是至关重要的。
该模型在研究百日咳中的宿主-寄生虫相互作用的相关性在于 PHAE 和 HBE 细胞与体内人呼吸道纤毛和杯状细胞之间存在显著的生理和形态相似性。这些细胞可以在体外增殖、分化,并表达与体内人呼吸道细胞相同的遗传特征。