Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
PLoS One. 2019 Nov 14;14(11):e0225224. doi: 10.1371/journal.pone.0225224. eCollection 2019.
The largest current disease-induced loss of vertebrate biodiversity is due to chytridiomycosis and despite the increasing understanding of the pathogenesis, knowledge unravelling the early host-pathogen interactions remains limited. Batrachochytrium dendrobatidis (Bd) zoospores attach to and invade the amphibian epidermis, with subsequent invasive growth in the host skin. Availability of an in vitro assay would facilitate in depth study of this interaction while reducing the number of experimental animals needed. We describe a fluorescent cell-based in vitro infection model that reproduces host-Bd interactions. Using primary keratinocytes from Litoria caerulea and the epithelial cell line A6 from Xenopus laevis, we reproduced different stages of host cell infection and intracellular growth of Bd, resulting in host cell death, a key event in chytridiomycosis. The presented in vitro models may facilitate future mechanistic studies of host susceptibility and pathogen virulence.
目前，最大的脊椎动物生物多样性疾病相关损失是由壶菌病引起的。尽管对发病机制的理解日益加深，但对于揭示宿主-病原体早期相互作用的知识仍然有限。两栖动物壶菌（Bd）游动孢子附着并侵入两栖动物表皮，随后在宿主皮肤中进行侵袭性生长。体外检测方法的出现将有助于深入研究这种相互作用，同时减少所需的实验动物数量。我们描述了一种基于荧光细胞的体外感染模型，可重现宿主与 Bd 的相互作用。使用来自蓝纹石龙子（Litoria caerulea）的原代角质形成细胞和非洲爪蟾（Xenopus laevis）的上皮细胞系 A6，我们重现了宿主细胞感染和 Bd 细胞内生长的不同阶段，导致宿主细胞死亡，这是壶菌病的一个关键事件。所提出的体外模型可能有助于未来对宿主易感性和病原体毒力的机制研究。
