Archives of Dermatological Research

, Volume 310, Issue 10, pp 849–856 | Cite as

Topical treatment with the bacterium-derived c-Met agonist InlB321/15 accelerates healing in the abrasion wound mouse model

  • Yaroslava M. Chalenko
  • Elena V. Sysolyatina
  • Konstantin A. Sobyanin
  • Marina R. Kapkaeva
  • Alexandra Lavrikova
  • Egor Kalinin
  • Olga N. Scheglovitova
  • Svetlana A. ErmolaevaEmail author
Concise Communication


Studies of factors affecting wound-healing rates are encouraged by a critical need for new treatments to manage an increasing burden of non-healing wounds. The InlB protein produced by the Gram-positive bacterium Listeria monocytogenes is an agonist of the tyrosine kinase receptor c-Met and a functional analog of the hepatocyte growth factor (HGF), which is a mammalian ligand of c-Met. The recombinant InlB321 protein, which is the c-Met-binding InlB domain (amino acids 31–321), was cloned from the L. monocytogenes serovar 4b clinical strain VIMHA015 and serovar 1/2a strain EGDe (InlB321/15 and InlB321/EGDe, respectively). Both InlB321 variants stimulated proliferation of endothelial HUVEC cells. InlB321/15 was more active in Erk1/2 phosphorylation assay, and more potent than InlB321/EGDe in the 2D-scratch wound-healing assay. Scratch closure reached 86%, 29% and 72% for InlB321/15, InlB321/EGDe and HGF, respectively, 72 h post-wounding (p < 0.05). Topically applied glycerol-mixed InlB321/15 (300 µg ml− 1) increased abrasion wound-healing rates in mice. The 50% wound closing time (CT50) was reduced by InlB321/15 (4.18 ± 0.91 days; CI: 3.05; 5.31) compared with control animals (5.51 ± 1.21 days; CI: 4.01; 7.01; p < 0.05). Taken together, obtained results suggested a potential of InlB321/15 as a means of accelerating wound healing.


Cutaneous wounds Wound healing c-Met Hepatocyte growth factor InlB 



The work was supported by Russian Science Foundation to S.E. (Grant no. 16-15-00091).

Compliance with ethical standards

Conflict of interest

There is no conflict of interests to disclose.

Ethical approval

Experiments on animals were conducted in accordance with the Russian Federation National Standard (GOST R52379-2005), directives of Ministry of Health of Russian Federation (No753n from 26.08.2010, No774н from 31.08.2010), and with the approval of the Biomedical Ethics Committee of Gamaleya Research Center of Epidemiology and Microbiology (No9 from 24.01.2017).


  1. 1.
    Appasamy R, Tanabe M, Murase N et al (1993) Hepatocyte growth factor, blood clearance, organ uptake, and biliary excretion in normal and partially hepatectomized rats. Lab Invest 68:270–276PubMedGoogle Scholar
  2. 2.
    Barrientos S, Stojadinovic O, Golinko MS et al (2008) Growth factors and cytokines in wound healing. Wound Repair Regen 16:585–601. CrossRefPubMedGoogle Scholar
  3. 3.
    Bierne H, Cossart P (2002) InlB, a surface protein of Listeria monocytogenes that behaves as an invasin and a growth factor. J Cell Sci 115:3357–3367PubMedGoogle Scholar
  4. 4.
    Bottaro DP, Rubin JS, Faletto DL et al (1991) Identification of the hepatocyte growth factor receptor as the c-met proto-oncogene product. Science 251:802–804. CrossRefPubMedGoogle Scholar
  5. 5.
    Buchstein N, Hoffmann D, Smola H et al (2009) Alternative proteolytic processing of hepatocyte growth factor during wound repair. Am J Pathol 174:2116–2128. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Bussolino F, Di Renzo MF, Ziche M et al (1992) Hepatocyte growth factor is a potent angiogenic factor which stimulates endothelial cell motility and growth. J Cell Biol 119:629–641. CrossRefPubMedGoogle Scholar
  7. 7.
    Chalenko YM, Sysolyatina EV, Kalinin EV et al (2017) Natural variants of Listeria monocytogenes internalin B with different ability to stimulate cell proliferation and cytoskeleton rearrangement in HEp-2 cells. Mol Genet Microbiol Virol 32:80–86. CrossRefGoogle Scholar
  8. 8.
    Chmielowiec J, Borowiak M, Morkel M et al (2007) c-Met is essential for wound healing in the skin. J Cell Biol 177:151–162. CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    D’Angelo F, Bernasconi E, Schäfer M et al (2013) Macrophages promote epithelial repair through hepatocyte growth factor secretion. Clin Exp Immunol 174:60–72. CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Ferraris DM, Gherardi E, Di Y et al (2010) Ligand-mediated dimerization of the met receptor tyrosine kinase by the bacterial invasion protein InlB. J Mol Biol 395:522–532. CrossRefPubMedGoogle Scholar
  11. 11.
    Ireton K, Payrastre B, Cossart P (1999) The Listeria monocytogenes protein InlB is an agonist of mammalian phosphoinositide 3-kinase. J Biol Chem 274:17025–17032. CrossRefPubMedGoogle Scholar
  12. 12.
    Kolditz F, Krausze J, Heinz DW et al (2014) Wound healing potential of a dimeric InlB variant analyzed by in vitro experiments on re-epithelialization of human skin models. Eur J Pharm Biopharm 86:277–283. CrossRefPubMedGoogle Scholar
  13. 13.
    Lecuit M, Ohayon H, Braun L et al (1997) Internalin of Listeria monocytogenes with an intact leucine-rich repeat region is sufficient to promote internalization. Infect Immun 65:5309–5319PubMedPubMedCentralGoogle Scholar
  14. 14.
    Matsumoto K, Funakoshi H, Takahashi H, Sakai K (2014) HGF-Met pathway in regeneration and drug discovery. Biomedicines 2:275–300. CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Miyazawa K, Tsubouchi H, Naka D et al (1989) Molecular cloning and sequence analysis of cDNA for human hepatocyte growth factor. Biochem Biophys Res Commun 163:967–973. CrossRefPubMedGoogle Scholar
  16. 16.
    Morishita R, Makino H, Aoki M et al (2011) Phase I/IIa clinical trial of therapeutic angiogenesis using hepatocyte growth factor gene transfer to treat critical limb ischemia. Arterioscler Thromb Vasc Biol 31:713–720. CrossRefPubMedGoogle Scholar
  17. 17.
    Nakamura T, Teramoto H, Ichihara A (1986) Purification and characterization of a growth factor from rat platelets for mature parenchymal hepatocytes in primary cultures. Proc Natl Acad Sci USA 83:6489–6493CrossRefGoogle Scholar
  18. 18.
    Niemann HH, Jäger V, Butler PJG et al (2007) Structure of the human receptor tyrosine kinase Met in complex with the Listeria invasion protein InlB. Cell 130:235–246. CrossRefPubMedGoogle Scholar
  19. 19.
    Scheglovitova ON, Romanov YA, Maksianina EV et al (2002) Herpes simplex type I virus infected human vascular endothelial cells induce the production of anti-viral and proinflammatory factors by peripheral blood leukocytes in vitro. Russ J Immunol 7:115–122PubMedGoogle Scholar
  20. 20.
    Shigematsu H, Yasuda K, Iwai T et al (2010) Randomized, double-blind, placebo-controlled clinical trial of hepatocyte growth factor plasmid for critical limb ischemia. Gene Ther 17:1152–1161. CrossRefPubMedGoogle Scholar
  21. 21.
    Sobyanin K, Sysolyatina E, Krivozubov M et al (2017) Naturally occurring InlB variants that support intragastric Listeria monocytogenes infection in mice. FEMS Microbiol Lett. CrossRefPubMedGoogle Scholar
  22. 22.
    Suzuki K, Takahashi K (2008) Regulation of lamellipodia formation and cell invasion by CLIP-170 in invasive human breast cancer cells. Biochem Biophys Res Commun 368:199–204. CrossRefPubMedGoogle Scholar
  23. 23.
    Toyoda M, Takayama H, Horiguchi N et al (2001) Overexpression of hepatocyte growth factor/scatter factor promotes vascularization and granulation tissue formation in vivo. FEBS Lett 509:95–100. CrossRefPubMedGoogle Scholar
  24. 24.
    Usatyuk PV, Fu P, Mohan V et al (2014) Role of c-Met/phosphatidylinositol 3-kinase (PI3k)/Akt signaling in Hepatocyte growth factor (HGF)-mediated lamellipodia formation, reactive oxygen species (ROS) generation, and motility of lung endothelial cells. J Biol Chem 289:13476–13491. CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Werner S, Grose R (2003) Regulation of wound healing by growth factors and cytokines. Physiol Rev 83:835–870. CrossRefPubMedGoogle Scholar
  26. 26.
    Yoshida S, Yamaguchi Y, Itami S et al (2003) Neutralization of hepatocyte growth factor leads to retarded cutaneous wound healing associated with decreased neovascularization and granulation tissue formation. J Invest Dermatol 120:335–343. CrossRefPubMedGoogle Scholar
  27. 27.
    Zarnegar R, DeFrances MC, Oliver L, Michalopoulos G (1990) Identification and partial characterization of receptor binding sites for HGF on rat hepatocytes. Biochem Biophys Res Commun 173:1179–1185. doi: S0006-291X(05)80910-6 [pii]CrossRefGoogle Scholar
  28. 28.
    Zioncheck TF, Richardson L, DeGuzman GG et al (1994) The pharmacokinetics, tissue localization, and metabolic processing of recombinant human hepatocyte growth factor after intravenous administration in rats. Endocrinology 134:1879–1887. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Yaroslava M. Chalenko
    • 1
  • Elena V. Sysolyatina
    • 1
  • Konstantin A. Sobyanin
    • 1
  • Marina R. Kapkaeva
    • 1
  • Alexandra Lavrikova
    • 1
  • Egor Kalinin
    • 1
  • Olga N. Scheglovitova
    • 1
  • Svetlana A. Ermolaeva
    • 1
    Email author
  1. 1.Gamaleya Research Center of Epidemiology and MicrobiologyMoscowRussia

Personalised recommendations