Invasion of Mycoplasma bovis into bovine synovial cells utilizing the clathrin-dependent endocytosis pathway

Nishi, Koji; Gondaira, Satoshi; Fujiki, Jumpei; Katagata, Michiko; Sawada, Chizuru; Eguchi, Ayako; Iwasaki, Tomohito; Iwano, Hidetomo; Higuchi, Hidetoshi

doi:10.1016/j.vetmic.2020.108956

Cited by 11 publications

(17 citation statements)

References 20 publications

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“…[72][73][74] A lipid raft-mediated endocytosis pathway was exploited by some pathogenic microorganisms, such as Chlamydia, for cell entry. Nishi K et al 50 reported that M. bovis can be internalized through a clathrin-dependent pathway or a clathrin-independent, lipid raft-mediated pathway, but the virulence factors responsible the mediated internalization process reunknown. In the current work, we hypothesized LppB could be beneficial for M. bovis to invade host cells through internalized pathways of clathrin-and lipid raft-mediated endocytosis.…”

Section: Discussionmentioning

confidence: 99%

“…Colocalization of LppB with clathrin and caveolin was examined by confocal microscopy. Endocytosis inhibition assays were performed as described previously with minor modifications 50 . To study the endocytic pathway involved in LppB internalization, two specific endocytic inhibitors were used 1 : pitstop‐2 (SML1169, Sigma), an inhibitor of clathrin‐mediated endocytosis 2 ; simvastatin (S6196, Sigma), an inhibitor of lipid raft‐mediated endocytosis.…”

Section: Methodsmentioning

confidence: 99%

“…Endocytosis inhibition assays were performed as described previously with minor modifications. 50 To study the endocytic pathway involved in LppB internalization, two specific endocytic inhibitors were used 1 : pitstop-2 (SML1169, Sigma), an inhibitor of clathrin-mediated endocytosis 2 ; simvastatin (S6196, Sigma), an inhibitor of lipid raft-mediated endocytosis. Cell membranes were stained with a red fluorochrome DiI (C1991S, Beyotime Biotechnology, Shanghai, China).…”

Section: Endocytic Pathways Of Lppbmentioning

confidence: 99%

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A genome‐wide transposon mutagenesis screening identifies LppB as a key factor associated with Mycoplasma bovis colonization and invasion into host cells

Lan,

Li,

Hao

et al. 2023

The FASEB Journal

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Mycoplasma spp., the smallest self‐replicating and genome‐reduced organisms, have raised a great concern in both the medical and veterinary fields due to their pathogenicity. The molecular determinants of these wall‐less bacterium efficiently use their limited genes to ensure successful infection of the host remain unclear. In the present study, we used the ruminant pathogen Mycoplasma bovis as a model to identify the key factors for colonization and invasion into host cells. We constructed a nonredundant fluorescent transposon mutant library of M. bovis using a modified transposon plasmid, and identified 34 novel adhesion‐related genes based on a high‐throughput screening approach. Among them, the ΔLppB mutant exhibited the most apparent decrease in adhesion to embryonic bovine lung (EBL) cells. The surface‐localized lipoprotein LppB, which is highly conserved in Mycoplasma species, was then confirmed as a key factor for M. bovis adhesion with great immunogenicity. LppB interacted with various components (fibronectin, vitronectin, collagen IV, and laminin) of host extracellular matrix (ECM) and promoted plasminogen activation through tPA to degrade ECM. The 439–502 amino acid region of LppB is a critical domain, and F465 and Y493 are important residues for the plasminogen activation activity. We further revealed LppB as a key factor facilitating internalization through clathrin‐ and lipid raft‐mediated endocytosis, which helps the Mycoplasma invade the host cells. Our study indicates that LppB plays a key role in Mycoplasma infection and is a potential new therapeutic and vaccine target for Mycoplasma species.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Endocytic Pathways Of Lppbmentioning

confidence: 99%

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A genome‐wide transposon mutagenesis screening identifies LppB as a key factor associated with Mycoplasma bovis colonization and invasion into host cells

Lan,

Li,

Hao

et al. 2023

The FASEB Journal

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“…Adhesion is an important step in M. bovis infection, the functional proteins (known as adhesins) are closely linked to the pathophysiological process ( 12 , 13 ). Traditionally, it is generally believed that M. bovis mainly parasitizes outside cells, but mounting evidence shows that M. bovis can invade host cells, and this mechanism is related to its distribution in host animals and immune escape ( 14 – 16 ). To date, M. bovis has been detected in neutrophils, macrophages, bronchial epithelial cells, hepatocytes and kidney cells of infected cattle ( 17 , 18 ).…”

Section: Adhesion Of M Bovis To Host Cellsmentioning

confidence: 99%

“…M. pneumoniae associates with integrin β1 on the surface of epithelial cells via interactions with surface-bound fibronectin and initiates signaling events that stimulate pathogen uptake into clathrin-coated vesicles and caveosomes ( 20 ). Some adhesins of M. bovis also take FN as the target protein, but whether M. bovis invades host cell also adopts the this mechanism remains to be further studied, although recent research shows that clathrin-dependent endocytosis is one of the major pathways by which M. bovis invades into synovial cells ( 14 ). The invasion of M. bovis into different cells may contribute to the spread of pathogens to various colonization sites, which is related to weakening the therapeutic effect of antibiotics and escaping from the immune system killing.…”

Section: Adhesion Of M Bovis To Host Cellsmentioning

confidence: 99%

Mycoplasma Bovis adhesins and their target proteins

Pan²,

Wu³

et al. 2022

Front. Immunol.

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Bovine mycoplasmosis is an important infectious disease of cattle caused by Mycoplasma bovis (M. bovis) which poses a serious threat to the breeding industry. Adhesin is involved in the initial process of M. bovis colonization, which is closely related to the infection, cell invasion, immune escape and virulence of this pathogenic microorganism. For the reason that M. bovis lacks a cell wall, its adhesin is predominantly located on the surface of the cell membrane. The adhesins of M. bovis are usually identified by adhesion and adhesion inhibition analysis, and more than 10 adhesins have been identified so far. These adhesins primarily bind to plasminogen, fibronectin, heparin and amyloid precursor-like protein-2 of host cells. This review aims to concisely summarize the current knowledge regarding the adhesins of M. bovis and their target proteins of the host cell. Additionally, the biological characteristics of the adhesin will be briefly analyzed.

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Effects of intra-articular inoculation with Mycoplasma bovis on immunological responses in calf joints

Nishi

Hirano

Sato

et al. 2022

Veterinary Immunology and Immunopathology

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Invasion of Mycoplasma bovis into bovine synovial cells utilizing the clathrin-dependent endocytosis pathway

Cited by 11 publications

References 20 publications

A genome‐wide transposon mutagenesis screening identifies LppB as a key factor associated with Mycoplasma bovis colonization and invasion into host cells

A genome‐wide transposon mutagenesis screening identifies LppB as a key factor associated with Mycoplasma bovis colonization and invasion into host cells

Mycoplasma Bovis adhesins and their target proteins

Effects of intra-articular inoculation with Mycoplasma bovis on immunological responses in calf joints

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