Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR, United States.
Department of Microbiology, College of Science, Oregon State University, Corvallis, OR, United States.
Front Cell Infect Microbiol. 2018 Apr 24;8:123. doi: 10.3389/fcimb.2018.00123. eCollection 2018.
subsp. () is a member of the non-tuberculous mycobacteria (NTM), and is a common cause of lung infection in patients with chronic NTM lung conditions. is an environmental bacterium believed to be transmitted from environmental sources. In this work we used a recently developed model in to ask whether can be transmitted from host-to-host, and the bacterial genes associated with host colonization. Infection of was carried out by placing the nematode in cultured with . Bacteria eliminated from the intestines of infected were used to infect naïve nematodes. In parallel experiments, to identify colonization associated genes, a transposon library of was screened for the ability to bind to HEp-2 mucosal cells. Thirty clones were identified and five selected clones with impaired adherence to HEp-2 epithelial cells were used to infect to determine the degree of colonization. It was determined that eliminated from infected were able to colonize a naïve with high efficiency. Thirty of the most adherence-deficient clones obtained from the HEp-2 cell screening were sequenced to identify the location of the transposon. Many of the genes associated with the bacterial cell wall synthesis were shown to be inactivated in the selected mutants. Five out of the 30 bacterial clones were then used to infect . All five mutants had impaired ability to colonize compared with the wild type bacteria (decrease of 1.5-2.0 logs, < 0.05). The limitation of this work is that the model can be used for initial screening, but other more complex systems should be used to confirm the findings. can be used as a model to test for adherence/colonization-associated virulence determinants. All the tested adherence-deficient clones that were examined had impaired ability to colonize the host , and some can be potentially used to prevent colonization.
亚种（）是一种非结核分枝杆菌（NTM），是慢性 NTM 肺部疾病患者肺部感染的常见原因。是一种环境细菌，据信是从环境来源传播的。在这项工作中，我们使用了最近在中开发的模型来询问是否可以从宿主传播到宿主，以及与宿主定植相关的细菌基因。通过将线虫放置在含有培养物中来进行感染。从感染线虫的肠道中消除的细菌用于感染未感染的线虫。在平行实验中，为了鉴定定植相关基因，对的转座子文库进行了筛选，以确定其与 HEp-2 粘膜细胞结合的能力。鉴定出 30 个克隆，并选择了 5 个对 HEp-2 上皮细胞粘附能力受损的克隆用于感染，以确定定植程度。结果表明，从感染线虫中消除的能够以高效率定植于未感染的线虫。从 HEp-2 细胞筛选中获得的 30 个最粘附缺陷克隆进行测序，以鉴定转座子的位置。许多与细菌细胞壁合成相关的基因被证明在选择的突变体中失活。然后，使用 30 个细菌克隆中的 5 个感染。与野生型细菌相比，所有 5 个突变体的定植能力都受损（减少 1.5-2.0 个对数级，<0.05）。这项工作的局限性在于该模型可用于初步筛选，但应使用其他更复杂的系统来确认研究结果。可以作为一种模型来测试粘附/定植相关毒力决定因素。所有经过测试的粘附缺陷克隆都表现出定植宿主的能力受损，其中一些可能可用于预防定植。
