Skip to main content
Log in

Cell immortalization enhances Listeria monocytogenes invasion

  • Original Investigations
  • Published:
Medical Microbiology and Immunology Aims and scope Submit manuscript

Abstract

Recent outbreaks of human listeriosis have emphasized the importance of food in the etiology of epidemic listeriosis, suggesting that the gastrointestinal tract is the natural site of entry for Listeria monocytogenes into the organism. L. monocytogenes invasion of finite cell lines derived from the porcine ileum exhibited a 100-fold lower penetration level, without any intracellular multiplication, when compared to CaCo-2 cells, a widely used in vitro model for L. monocytogenes invasion. Same results were obtained with both pig kidney primary cells and mouse kidney finite cell lines. To demonstrate that cell immortalization enhances L. monocytogenes invasion, finite cell lines from porcine ileum and from murine kidney were immortalized by Simian virus 40 (SV40) large T oncogene. Unlike their untransformed counterparts, the immortal cells obtained were invaded by L. monocytogenes, as observed for CaCo-2 cells as well as for spontaneously immortal human (HeLa) and murine (3T3) cell lines. Extensive electron microscopy examinations of porcine epithelioid cells infected by L. monocytogenes showed numerous bacteria within the immortal cells, whereas neither intracellular bacteria nor any bacterial antigen were revealed inside finite cell lines. These data suggested that L. monocytogenes were not destroyed inside finite cell lines but only poorly entered the finite or primary cells. Speculating that L. monocytogenes invasion is under control of differentiation or proliferation of the cells, only an enterocyte subset at a defined state of differentiation or expressing particular receptors could be invaded in vivo.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Audurier A, Pardon P, Marly J, Lantier F, Loulergue J (1981) Mesure de la virulence chez la souris de différentes bactéries appartenant au genre Listeria. Ann (Inst. Pasteur) Immunol 132D:191–200

    Google Scholar 

  2. Brown KN, Percival A (1978) Penetration of antimicrobials into tissue culture cells and leukocytes. Scand J Infect Dis [Suppl] 14:251–260

    Google Scholar 

  3. Bubert A, Kuhn M, Goebel W, Kohler S (1992) Structural and functional properties of the p60 proteins from different Listeria species. J Bacteriol 174:8166–8171

    Google Scholar 

  4. Chakraborty T, Goebel W (1988) Recent developments in the study of virulence in Listeria monocytogenes. Curr Top Microbiol Immunol 138:41–58

    Google Scholar 

  5. Chen TR (1977) In situ detection of Mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. Exp Cell Res 104:255–262

    Google Scholar 

  6. Cloeckaert A, Zygmunt M, De Wergifosse P, Dubray G, Limet JN (1992) Demonstration of peptidoglycan-associated Brucella outermembrane proteins by use of monoclonal antibodies. J Gen Microbiol 138:1543–1550

    Google Scholar 

  7. Eveillard M, Fourel V, Barc MC, Kerneis S, Coconnier MH, Karjalainen T, Bourlioux P, Servin AL (1993) Identification and characterization of adhesive factors of Clostridium difficile involved in adhesion to human colonic enterocyte-like caco-2 and mucus-secreting HT29 cells in culture. Mol Microbiol 7:371–381

    Google Scholar 

  8. Falkow S (1991) Bacterial entry into eukaryotic cells. Cell 65:1099–1102

    Google Scholar 

  9. Falkow S, Isberg RR, Portnoy DA (1992) The interaction of bacteria with mammalian cells. Annu Rev Cell Biol 8:333–363

    Google Scholar 

  10. Fantini J (1992) Differentiation in human colon tumor cells. Importance of autocrine growth factors. Cancer J 5:78–83

    Google Scholar 

  11. Freshney RI (1987) Culture of animal cells. A manual of basic technique. Liss, New York

    Google Scholar 

  12. Gaillard JL, Berche P, Mounier J, Richard S, Sansonetti P (1978) In vitro model of penetration and intracellular growth of Listeria monocytogenes in the human enterocyte-like cell line CaCo-2. Infect Immun 55:2822–2829

    Google Scholar 

  13. Gaillard JL, Berche P, Frehel C, Gouin E, Cossart P (1991) Entry of L. monocytogenes into cells is mediated by internalin, a repeat protein reminiscent of surface antigens from Grampostivie Cocci. Cell 65:1127–1141

    Google Scholar 

  14. Galan JE, Pace J, Hayman MJ (1992) Involvement of the epidermal growth factor receptor in the invasion of cultured mammalian cells by Salmonella typhimurium. Nature 357:588–589

    Google Scholar 

  15. Gilbert SF, Migeon BR (1975) d-Valine as a selective agent for normal human and rodent epithelial cells in culture. Cell 5:11–17

    Google Scholar 

  16. Goulet V, Lepoutre A, Rocourt J, Courtieu AL, Dehaumont P, Veit P (1993) Epidémie de listériose en France. Bilan final et résultats de l'enquête épidémiologique. Bull Epidemiol Heb 4:13–14

    Google Scholar 

  17. Havell EA (1986) Synthesis and secretion of interferon by murine fibroblasts in response to intracellular Listeria monocytogenes. Infect Immun 54:787–792

    Google Scholar 

  18. Jones D (1990) Foodborne Listeriosis. Lancet 336:1171–1174

    Google Scholar 

  19. Kaeffer B, Bottreau E, Velge P, Pardon P (1993) Epithelioid and fibroblastic cell lines derived from the ileum of an adult histocompatible miniature boar (d/d haplotype) and immortalized by SV40 plasmid. Eur J Cell Biol 65:152–162

    Google Scholar 

  20. Kerneis S, Bilge SS, Fourel V, Chauviere G, Coconnier MH, Servin AL (1991) Use of purified f1845 fimbrial adhesin to study localization and expression of receptors for diffusely adhering Escherichia coli during enterocytic differentiation of human colon carcinoma cell lines HT-29 and CaCo-2 in culture. Infect Immun 59:4013–4018

    Google Scholar 

  21. Köhler G, Milstein C (1975) Continuous culture fused cells secreting antibody of predifined specificity. Nature 26:495–498

    Google Scholar 

  22. Kuhn M, Kathariou S, Goebel W (1988) Hemolysin supports survival but not entry on the intracellular bacterium Listeria monocytogenes. Infect Immun 56:79–82

    Google Scholar 

  23. Lefford MJ, Warner S, Amell L (1979) Listeria pneumonitis: influence of route immunization on resistance to airborne infection. Infect Immun 25:672–679

    Google Scholar 

  24. Linder S, Marshal H (1990) Immortalization of primary cells by DNA tumor viruses. Exp Cell Res 191:1–7

    Google Scholar 

  25. Louvard D, Kedinger M, Hauri HP (1992) The differentiating intestinal epithelial cell: establishment and maintenance of functions through interactions between cellular structures. Annu Rev Cell Biol 8:157–195

    Google Scholar 

  26. MacDonald TT, Carter PB (1980) Cell-mediated immunity to intestinal infection. Infect Immun 28:516–523

    Google Scholar 

  27. Marco AJ, Domingo M, Prats M, Briones V, Pumarola M, Dominguez L (1991) Pathogenesis of lymphoid lesions in murine experimental listeriosis. J Comp Pathol 105:1–15

    Google Scholar 

  28. Meyer DH, Bunduki M, Beliveau CM, Donnelly CW (1992) Differences in invasion and adherence of Listeria monocytogenes with mammalian gut cells. Food Microbiol 9:115–126

    Google Scholar 

  29. Morris A, Steinberg ML, Defendi V (1985) Keratin gene expression in simian virus 40-transformed human keratinocytes. Proc Natl Acad Sci USA 82:8498–8502

    Google Scholar 

  30. Payrastre B (1992) Cytosquelette, phospho-inositides et transduction du signal, médecine/sciences 8:127–133

    Google Scholar 

  31. Portnoy DA, Chakraborty T, Goebel W, Cossart P (1992) Molecular determinants of Listeria monocytogenes pathogenesis. Infect Immun 60:1263–1267

    Google Scholar 

  32. Portnoy DA, Jacks PS, Hinrichs DJ (1988) Role of hemolysin for the intracellular growth of L. monocytogenes. J Exp Med 167:1459–1471

    Google Scholar 

  33. Premaratne RJ, Lin WJ, Johnson EA (1991) Development of an improved chemically defined minimal medium for Listeria monocytogenes. Appl Environ Microbiol 57:3046–3048

    Google Scholar 

  34. Racz P, Tenner K, Mérö E (1972) Experimental Listeria enteritis I. An electron microscopic study of the epithelial phase in experimental Listeria infection. Lab Invest 26:694–700

    Google Scholar 

  35. Rosenshine I, Donnenberg MS, Kaper JB, Finlay BB (1992) Signal transduction between enteropathogenic Escherichia coli (EPEC) an epithelial cells — EPEC induces tyrosine phosphorylation of host cell proteins to initiate cytoskeletal rearrangement and bacterial uptake. EMBO J 11:3551–3560

    Google Scholar 

  36. Rosenshine I, Duronio V, Finlay BB (1992) Tyrosine protein kinase inhibitors block invasion-promoted bacterial uptake by epithelial cells. Infect Immun 60:2211–2217

    Google Scholar 

  37. Ryser ET, Marth EH (1991) Listeria monocytogenes: characteristics and classification in Listeria, Listeriosis, and food safety. Food science and technology serie 46:1–21

    Google Scholar 

  38. Schlech WF (1984) New perspectives on the gastrointestinal mode of transmission in invasive Listeria monocytogenes infection. J Invest Med 7:321–324

    Google Scholar 

  39. Southern PJ, Berg P (1982) Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet 1:327–341

    Google Scholar 

  40. Tabouret M, Derycke J, Dubray G (1992) Analysis of surface proteins of Listeria in relation to species, serovar and pathogenicity. J Gen Microbiol 138:743–753

    Google Scholar 

  41. Tang P, Foubister V, Pucciarelli MG, Finlay BB (1993) Methods to study bacterial invasion. J Microbiol Methods 240:227–238

    Google Scholar 

  42. Tilney LG, Portnoy DA (1989) Actin filaments and the growth, movement, and spread of the intracellular bacterial parasite, Listeria monocytogenes. J Cell Biol 109:1597–1608

    Google Scholar 

  43. Tilney LG, Derosier DJ, Tilney MS (1992) How Listeria exploits host cell actin to form its own cytoskeleton. 1. Formation of a tail and how that tail might be involved in movement. J Cell Biol 118:71–81

    Google Scholar 

  44. Velge P, Ouaissi MA, Cornette J, Afchain D, Capron A (1988) Identification and isolation of Trypanosoma cruzi trypomastigote collagen binding proteins: possible role in cell-parasite interaction. Parasitology 97:255–268

    Google Scholar 

  45. Velge P, Bottreau E, Kaeffer B, Yurdusev N, Pardon P, Van Langendonck N Protein Tyrosine Kinase inhibitors block the entries of Listeria monocytogenes and Listeria ivanovii into epithelial cells. Microbial Pathogenesis (in press)

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Velge, P., Bottreau, E., Kaeffer, B. et al. Cell immortalization enhances Listeria monocytogenes invasion. Med Microbiol Immunol 183, 145–158 (1994). https://doi.org/10.1007/BF00196049

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00196049

Keywords

Navigation