Department of Biochemistry, Genetics and Microbiology, University of Pretoria, Pretoria, South Africa.
Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, Indiana, United States of America.
PLoS One. 2019 Jul 29;14(7):e0220321. doi: 10.1371/journal.pone.0220321. eCollection 2019.
Listeria monocytogenes is an intracellular foodborne pathogen that employs a number of strategies to survive challenging gastrointestinal conditions. It proliferates in the gut and subsequently causes listeriosis in high-risk individuals. Therefore, inhibition of its adherence to the intestinal receptors is crucial in controlling its infection. In this study, the effect of our previously developed recombinant Lactobacillus casei strain expressing invasion protein, Internalin AB of L. monocytogenes (LbcInlAB) on epithelial infection processes of the latter under simulated intestinal conditions was investigated.
The confluent Caco-2 cell monolayer was pre-exposed to different L. casei strains at a multiplicity of exposure (MOE) of 10 for various periods before infection with L. monocytogenes at a multiplicity of infection (MOI) of 10 under simulated intestinal conditions. Subsequently, L. monocytogenes adhesion, invasion, and translocation, cytotoxicity and impact on tight junction integrity of the Caco-2 cells were analyzed.
Under the simulated gastrointestinal condition, LbcInlAB showed a significant increase (p<0.0001) in adherence to, invasion and translocation through the Caco-2 cells when compared with the wild type strain. Although LbcInlAB strain exhibited enhanced inhibition of L. monocytogenes, it was not able to displace L. monocytogenes cells already attached to the monolayer. Additionally, pre-exposure to LbcInlAB reduced L. monocytogenes-mediated cytotoxicity and protected the tight junction barrier function.
The recombinant L. casei expressing InlAB shows potential for use as a prophylactic intervention strategy for targeted control of L. monocytogenes during the intestinal phase of infection.
单核细胞增生李斯特菌是一种食源性胞内病原体,它采用多种策略来应对挑战性的胃肠道环境。它在肠道中增殖,然后在高危人群中引起李斯特菌病。因此,抑制其与肠道受体的黏附对于控制其感染至关重要。在这项研究中,我们研究了先前开发的表达侵袭蛋白 Internalin AB 的重组干酪乳杆菌菌株对模拟肠道条件下后者对上皮感染过程的影响。
在模拟肠道条件下,用单核细胞增生李斯特菌的感染复数(MOI)为 10 感染 Caco-2 细胞单层之前,用不同的干酪乳杆菌菌株以 10 的暴露多重数(MOE)预先暴露于 Caco-2 细胞单层不同时间。随后,分析单核细胞增生李斯特菌的黏附、侵袭和易位、细胞毒性以及对 Caco-2 细胞紧密连接完整性的影响。
在模拟胃肠道条件下,与野生型菌株相比,LbcInlAB 对 Caco-2 细胞的黏附、侵袭和易位显著增加(p<0.0001)。尽管 LbcInlAB 菌株对单核细胞增生李斯特菌的抑制作用增强,但它不能取代已附着在单层上的单核细胞增生李斯特菌细胞。此外,LbcInlAB 的预先暴露降低了单核细胞增生李斯特菌介导的细胞毒性并保护了紧密连接屏障功能。
表达 InlAB 的重组干酪乳杆菌具有作为预防干预策略的潜力,可用于在感染的肠道阶段靶向控制单核细胞增生李斯特菌。