Natural variants of Listeria monocytogenes internalin B with different ability to stimulate cell proliferation and cytoskeleton rearrangement in HEp-2 cells

Chalenko, Ya. M.; Sysolyatina, Elena V.; Kalinin, Egor; Sobyanin, K. A.; Sа, Ermolaeva

doi:10.3103/s0891416817020021

Cited by 7 publications

(11 citation statements)

References 16 publications

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“…This function might be important for the role of interactions between gC1q-R and idInlB in L. monocytogenes active cell invasion. Results obtained in this work are in line with our previous results demonstrating differences in lamellipodia and filopodia formation induced by different idInlB isoforms in HEp-2 cells [65]. The known role of InlB as not only an invasion factor but also as a endocytosis activating factor [66] provides a direction for further studies on functional roles of its phylogenetically defined variability.…”

Section: Discussionsupporting

confidence: 90%

Phylogenetically Defined Isoforms of Listeria monocytogenes Invasion Factor InlB Differently Activate Intracellular Signaling Pathways and Interact with the Receptor gC1q-R

Chalenko

Kalinin

Marchenkov

et al. 2019

IJMS

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The pathogenic Gram-positive bacterium Listeria monocytogenes has been evolving into a few phylogenetic lineages. Phylogenetically defined substitutions were described in the L. monocytogenes virulence factor InlB, which mediates active invasion into mammalian cells via interactions with surface receptors c-Met and gC1q-R. InlB internalin domain (idInlB) is central to interactions with c-Met. Here we compared activity of purified recombinant idInlB isoforms characteristic for L. monocytogenes phylogenetic lineage I and II. Size exclusion chromatography and intrinsic fluorescence were used to characterize idInlBs. Western blotting was used to study activation of c-Met-dependent MAPK- and PI3K/Akt-pathways. Solid-phase microplate binding and competition assay was used to quantify interactions with gCq1-R. Isogenic recombinant L. monocytogenes strains were used to elucidate the input of idInlB isoforms in HEp-2 cell invasion. Physicochemical parameters of idInlB isoforms were similar but not identical. Kinetics of Erk1/2 and Akt phosphorylation in response to purified idInlBs was lineage specific. Lineage I but not lineage II idInlB specifically bound gC1q-R. Antibody against gC1q-R amino acids 221–249 inhibited invasion of L. monocytogenes carrying lineage I but not lineage II idInlB. Taken together, obtained results suggested that phylogenetically defined substitutions in idInlB provide functional distinctions and might be involved in phylogenetically determined differences in virulence potential.

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Section: Discussionsupporting

confidence: 90%

Phylogenetically Defined Isoforms of Listeria monocytogenes Invasion Factor InlB Differently Activate Intracellular Signaling Pathways and Interact with the Receptor gC1q-R

Chalenko

Kalinin

Marchenkov

et al. 2019

IJMS

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“…Two male rabbits (weighing 2.5 kg) were immunized with the recombinant protein InlB321/15 according to the scheme ( Table 1 ). InlB321/15 was purified from the recombinant E. coli BL21::pET28b::inlBallele9 strain as described previously [ 94 ]. Briefly, after the first immunization with the protein applied intravenously, subcutaneously and intramuscularly injections were performed on the 4th, 8th and 10th weeks.…”

Section: Methodsmentioning

confidence: 99%

Listeria monocytogenes Infection of Bat Pipistrellus nathusii Epithelial cells Depends on the Invasion Factors InlA and InlB

Povolyaeva

Chalenko²,

Kalinin³

et al. 2020

Pathogens

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L. monocytogenes is a widespread facultative intracellular pathogen. The range of natural hosts that supporting L. monocytogenes persistence in the environment has not been fully established yet. In this study, we were interested in the potential of L. monocytogenes to infect cells of bats, which are being increasingly recognized as a reservoir for microorganisms that are pathogenic to humans and domestic animals. A stable epithelial cell line was developed from the kidneys of Pipistrellus nathusii, a small bat widely distributed across Europe. The wild-type L. monocytogenes strain EGDe infected this cell line with an invasion efficiency of 0.0078 ± 0.0009%. Once it entered bat cells, L. monocytogenes doubled within about 70 minutes. When L. monocytogenes lacked either of the major invasion factors, InlA and InlB, invasion efficiency decreased by a factor of 10 and 25 respectively (p < 0.000001). The obtained results suggest that bat epithelial cells are susceptible to L. monocytogenes infection and that L. monocytogenes invasion of bat cells depends on the major invasion factors InlA and InlB. These results constitute the first report on in vitro studies of L. monocytogenes infection in bats.

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“…InlB321/15 was purified via His-tag with Dynabeads TM (Invitrogen TM , Thermo Fisher Scientific) from the recombinant E. coli BL21::pET28b::inlBallele9 strain as described [13]. The pET26b::inlBallele9 plasmid carried the InlB internalin domain encoding part of inlB gene from the virulent L. monocytogenes strain VIMHA015 isolated from a stillborn baby with listeriosis in Russia [13]. Protein purity was confirmed with SDS-PAGE (data not shown).…”

Section: Methodsmentioning

confidence: 99%

“…InlB/HGFR interactions promote cell invasion, causing bacterial infection [9,11]. Purified InlB acts as a growth factor and stimulates cell proliferation and migration [11,12,13]. The InlB leucine-repeat rich (LRR)-domain flanked by specific N -cap and immunoglobulin-like domains, which are known together as an internalin domain or InlB321, is sufficient to activate HGFR controlled cascades [14,15].…”

Section: Introductionmentioning

confidence: 99%

“…Bacterial proteins have comparative advantages over recombinant human growth factors in regard to the accessibility, efficiency, and simplicity of production. Several studies have demonstrated the potential for InlB321 variants derived from different L. monocytogenes strains to stimulate the proliferation and motility of primary lung endothelial cells, keratinocytes, and Human Umblical Vein Endothelial Cells [13,14,15,16,17]. Recently, we showed that the variant InlB321 from L. monocytogenes strain VIMHA015 (InlB321/15) accelerated abrasion wound healing in mice [16].…”

Section: Introductionmentioning

confidence: 99%

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Hepatoprotective Activity of InlB321/15, the HGFR Ligand of Bacterial Origin, in CCI4-Induced Acute Liver Injury Mice

et al. 2019

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HGF (hepatocyte growth factor)/HGFR (HGF receptor) signaling pathway is a key pathway in liver protection and regeneration after acute toxic damage. Listeria monocytogenes toxin InlB contains a HGFR-interacting domain and is a functional analog of HGF. The aim of this work was to evaluate the hepatoprotective activity of the InlB HGFR-interacting domain. The recombinant HGFR-interacting domain InlB321/15 was purified from E. coli. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test was used to measure InlB321/15 mitogenic activity in HepG2 cells. Activation of MAPK- and PI3K/Akt-pathways was tracked with fluorescent microscopy, Western blotting, and ELISA. To evaluate hepatoprotective activity, InlB321/15 and recombinant human HGF (rhHGF) were intravenously injected at the same concentration of 2 ng·g−1 to BALB/c mice 2 h before liver injury with CCl4. InlB321/15 caused dose-dependent activation of MAPK- and PI3K/Akt-pathways and correspondent mitogenic effects. Both InlB321/15 and rhHGF improved macroscopic liver parameters (liver mass was 1.51, 1.27 and 1.15 g for the vehicle, InlB321/15 and rhHGF, respectively, p < 0.05), reduced necrosis (24.0%, 16.18% and 21.66% of the total area for the vehicle, InlB321/15 and rhHGF, respectively, p < 0.05). Obtained data suggest that InlB321/15 is a promising candidate for a tissue repair agent.

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Natural variants of Listeria monocytogenes internalin B with different ability to stimulate cell proliferation and cytoskeleton rearrangement in HEp-2 cells

Cited by 7 publications

References 16 publications

Phylogenetically Defined Isoforms of Listeria monocytogenes Invasion Factor InlB Differently Activate Intracellular Signaling Pathways and Interact with the Receptor gC1q-R

Phylogenetically Defined Isoforms of Listeria monocytogenes Invasion Factor InlB Differently Activate Intracellular Signaling Pathways and Interact with the Receptor gC1q-R

Listeria monocytogenes Infection of Bat Pipistrellus nathusii Epithelial cells Depends on the Invasion Factors InlA and InlB

Hepatoprotective Activity of InlB321/15, the HGFR Ligand of Bacterial Origin, in CCI4-Induced Acute Liver Injury Mice

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