Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1992 Nov;60(11):4925–4931. doi: 10.1128/iai.60.11.4925-4931.1992

Laminin on Toxoplasma gondii mediates parasite binding to the beta 1 integrin receptor alpha 6 beta 1 on human foreskin fibroblasts and Chinese hamster ovary cells.

G C Furtado 1, Y Cao 1, K A Joiner 1
PMCID: PMC258249  PMID: 1399003

Abstract

We investigated the role of parasite-bound laminin and the host cell beta 1 integrin receptors for this extracellular matrix protein in Toxoplasma gondii binding to fibroblasts. Laminin but not fibronectin was detected on extracellular tachyzoites by immunofluorescence and immunoblotting. Binding of parasites to CHO cells was inhibited by polyclonal antibodies to laminin and by a monoclonal antibody directed against the globular carboxyl-terminal portion of the long arm of laminin (at or near the suggested ligand-binding sites for alpha 3 beta 1 and alpha 6 beta 1), but not by a monoclonal antibody directed against the lateral short arms of laminin near the cross region of the molecule. Antibodies to the alpha 6 but not the alpha 2, alpha 3, or alpha 5 chains of the beta 1 family of integrins blocked parasite attachment to human foreskin fibroblasts and CHO cells. Attachment of T. gondii to cells via laminin on the parasite surface and laminin receptors on the mammalian cell is consistent with the capacity of the parasite to invade almost all nucleated cells.

Full text

PDF
4931

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aumailley M., Nurcombe V., Edgar D., Paulsson M., Timpl R. The cellular interactions of laminin fragments. Cell adhesion correlates with two fragment-specific high affinity binding sites. J Biol Chem. 1987 Aug 25;262(24):11532–11538. [PubMed] [Google Scholar]
  2. Baughn R. E. Role of fibronectin in the pathogenesis of syphilis. Rev Infect Dis. 1987 Jul-Aug;9 (Suppl 4):S372–S385. doi: 10.1093/clinids/9.supplement_4.s372. [DOI] [PubMed] [Google Scholar]
  3. Bednarczyk J. L., McIntyre B. W. A monoclonal antibody to VLA-4 alpha-chain (CDw49d) induces homotypic lymphocyte aggregation. J Immunol. 1990 Feb 1;144(3):777–784. [PubMed] [Google Scholar]
  4. Buck C. A., Horwitz A. F. Cell surface receptors for extracellular matrix molecules. Annu Rev Cell Biol. 1987;3:179–205. doi: 10.1146/annurev.cb.03.110187.001143. [DOI] [PubMed] [Google Scholar]
  5. Casta e Silva Filho F., de Souza W., Lopes J. D. Presence of laminin-binding proteins in trichomonads and their role in adhesion. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8042–8046. doi: 10.1073/pnas.85.21.8042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Charonis A. S., Skubitz A. P., Koliakos G. G., Reger L. A., Dege J., Vogel A. M., Wohlhueter R., Furcht L. T. A novel synthetic peptide from the B1 chain of laminin with heparin-binding and cell adhesion-promoting activities. J Cell Biol. 1988 Sep;107(3):1253–1260. doi: 10.1083/jcb.107.3.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clément B., Segui-Real B., Savagner P., Kleinman H. K., Yamada Y. Hepatocyte attachment to laminin is mediated through multiple receptors. J Cell Biol. 1990 Jan;110(1):185–192. doi: 10.1083/jcb.110.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dawson J. R., Ellen R. P. Tip-oriented adherence of Treponema denticola to fibronectin. Infect Immun. 1990 Dec;58(12):3924–3928. doi: 10.1128/iai.58.12.3924-3928.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elices M. J., Osborn L., Takada Y., Crouse C., Luhowskyj S., Hemler M. E., Lobb R. R. VCAM-1 on activated endothelium interacts with the leukocyte integrin VLA-4 at a site distinct from the VLA-4/fibronectin binding site. Cell. 1990 Feb 23;60(4):577–584. doi: 10.1016/0092-8674(90)90661-w. [DOI] [PubMed] [Google Scholar]
  10. Finlay B. B., Falkow S. Common themes in microbial pathogenicity. Microbiol Rev. 1989 Jun;53(2):210–230. doi: 10.1128/mr.53.2.210-230.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fuhrman S. A., Joiner K. A. Toxoplasma gondii: mechanism of resistance to complement-mediated killing. J Immunol. 1989 Feb 1;142(3):940–947. [PubMed] [Google Scholar]
  12. Furtado G. C., Slowik M., Kleinman H. K., Joiner K. A. Laminin enhances binding of Toxoplasma gondii tachyzoites to J774 murine macrophage cells. Infect Immun. 1992 Jun;60(6):2337–2342. doi: 10.1128/iai.60.6.2337-2342.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gehlsen K. R., Dickerson K., Argraves W. S., Engvall E., Ruoslahti E. Subunit structure of a laminin-binding integrin and localization of its binding site on laminin. J Biol Chem. 1989 Nov 15;264(32):19034–19038. [PubMed] [Google Scholar]
  14. Gehlsen K. R., Dillner L., Engvall E., Ruoslahti E. The human laminin receptor is a member of the integrin family of cell adhesion receptors. Science. 1988 Sep 2;241(4870):1228–1229. doi: 10.1126/science.2970671. [DOI] [PubMed] [Google Scholar]
  15. Gehlsen K. R., Sriramarao P., Furcht L. T., Skubitz A. P. A synthetic peptide derived from the carboxy terminus of the laminin A chain represents a binding site for the alpha 3 beta 1 integrin. J Cell Biol. 1992 Apr;117(2):449–459. doi: 10.1083/jcb.117.2.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Graf J., Iwamoto Y., Sasaki M., Martin G. R., Kleinman H. K., Robey F. A., Yamada Y. Identification of an amino acid sequence in laminin mediating cell attachment, chemotaxis, and receptor binding. Cell. 1987 Mar 27;48(6):989–996. doi: 10.1016/0092-8674(87)90707-0. [DOI] [PubMed] [Google Scholar]
  17. Hall D. E., Frazer K. A., Hann B. C., Reichardt L. F. Isolation and characterization of a laminin-binding protein from rat and chick muscle. J Cell Biol. 1988 Aug;107(2):687–697. doi: 10.1083/jcb.107.2.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hall D. E., Reichardt L. F., Crowley E., Holley B., Moezzi H., Sonnenberg A., Damsky C. H. The alpha 1/beta 1 and alpha 6/beta 1 integrin heterodimers mediate cell attachment to distinct sites on laminin. J Cell Biol. 1990 Jun;110(6):2175–2184. doi: 10.1083/jcb.110.6.2175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hemler M. E. VLA proteins in the integrin family: structures, functions, and their role on leukocytes. Annu Rev Immunol. 1990;8:365–400. doi: 10.1146/annurev.iy.08.040190.002053. [DOI] [PubMed] [Google Scholar]
  20. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  21. Ignatius M. J., Reichardt L. F. Identification of a neuronal laminin receptor: an Mr 200K/120K integrin heterodimer that binds laminin in a divalent cation-dependent manner. Neuron. 1988 Oct;1(8):713–725. doi: 10.1016/0896-6273(88)90170-5. [DOI] [PubMed] [Google Scholar]
  22. Isberg R. R. Discrimination between intracellular uptake and surface adhesion of bacterial pathogens. Science. 1991 May 17;252(5008):934–938. doi: 10.1126/science.1674624. [DOI] [PubMed] [Google Scholar]
  23. Isberg R. R., Leong J. M. Multiple beta 1 chain integrins are receptors for invasin, a protein that promotes bacterial penetration into mammalian cells. Cell. 1990 Mar 9;60(5):861–871. doi: 10.1016/0092-8674(90)90099-z. [DOI] [PubMed] [Google Scholar]
  24. Joiner K. A. Complement evasion by bacteria and parasites. Annu Rev Microbiol. 1988;42:201–230. doi: 10.1146/annurev.mi.42.100188.001221. [DOI] [PubMed] [Google Scholar]
  25. Joiner K. A., Fuhrman S. A., Miettinen H. M., Kasper L. H., Mellman I. Toxoplasma gondii: fusion competence of parasitophorous vacuoles in Fc receptor-transfected fibroblasts. Science. 1990 Aug 10;249(4969):641–646. doi: 10.1126/science.2200126. [DOI] [PubMed] [Google Scholar]
  26. Kawamoto F., Kumada N. Fluorescent probes for detection of protozoan parasites. Parasitol Today. 1987 Sep;3(9):284–286. doi: 10.1016/0169-4758(87)90110-4. [DOI] [PubMed] [Google Scholar]
  27. Kramer R. H., Cheng Y. F., Clyman R. Human microvascular endothelial cells use beta 1 and beta 3 integrin receptor complexes to attach to laminin. J Cell Biol. 1990 Sep;111(3):1233–1243. doi: 10.1083/jcb.111.3.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  29. Languino L. R., Gehlsen K. R., Wayner E., Carter W. G., Engvall E., Ruoslahti E. Endothelial cells use alpha 2 beta 1 integrin as a laminin receptor. J Cell Biol. 1989 Nov;109(5):2455–2462. doi: 10.1083/jcb.109.5.2455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lopes J. D., dos Reis M., Brentani R. R. Presence of laminin receptors in Staphylococcus aureus. Science. 1985 Jul 19;229(4710):275–277. doi: 10.1126/science.3160113. [DOI] [PubMed] [Google Scholar]
  31. Lotz M. M., Korzelius C. A., Mercurio A. M. Human colon carcinoma cells use multiple receptors to adhere to laminin: involvement of alpha 6 beta 4 and alpha 2 beta 1 integrins. Cell Regul. 1990 Feb;1(3):249–257. doi: 10.1091/mbc.1.3.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Malinoff H. L., Wicha M. S. Isolation of a cell surface receptor protein for laminin from murine fibrosarcoma cells. J Cell Biol. 1983 May;96(5):1475–1479. doi: 10.1083/jcb.96.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mercurio A. M., Shaw L. M. Macrophage interactions with laminin: PMA selectively induces the adherence and spreading of mouse macrophages on a laminin substratum. J Cell Biol. 1988 Nov;107(5):1873–1880. doi: 10.1083/jcb.107.5.1873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mollenhauer J., von der Mark K. Isolation and characterization of a collagen-binding glycoprotein from chondrocyte membranes. EMBO J. 1983;2(1):45–50. doi: 10.1002/j.1460-2075.1983.tb01378.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Neugebauer K. M., Reichardt L. F. Cell-surface regulation of beta 1-integrin activity on developing retinal neurons. Nature. 1991 Mar 7;350(6313):68–71. doi: 10.1038/350068a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nichols B. A., O'Connor G. R. Penetration of mouse peritoneal macrophages by the protozoon Toxoplasma gondii. New evidence for active invasion and phagocytosis. Lab Invest. 1981 Apr;44(4):324–335. [PubMed] [Google Scholar]
  37. Ouaissi M. A., Afchain D., Capron A., Grimaud J. A. Fibronectin receptors on Trypanosoma cruzi trypomastigotes and their biological function. Nature. 1984 Mar 22;308(5957):380–382. doi: 10.1038/308380a0. [DOI] [PubMed] [Google Scholar]
  38. Ouaissi M. A. Role of the RGD sequence in parasite adhesion to host cells. Parasitol Today. 1988 Jun;4(6):169–173. doi: 10.1016/0169-4758(88)90153-6. [DOI] [PubMed] [Google Scholar]
  39. Proctor R. A. The staphylococcal fibronectin receptor: evidence for its importance in invasive infections. Rev Infect Dis. 1987 Jul-Aug;9 (Suppl 4):S335–S340. doi: 10.1093/clinids/9.supplement_4.s355. [DOI] [PubMed] [Google Scholar]
  40. Pulido R., Elices M. J., Campanero M. R., Osborn L., Schiffer S., García-Pardo A., Lobb R., Hemler M. E., Sánchez-Madrid F. Functional evidence for three distinct and independently inhibitable adhesion activities mediated by the human integrin VLA-4. Correlation with distinct alpha 4 epitopes. J Biol Chem. 1991 Jun 5;266(16):10241–10245. [PubMed] [Google Scholar]
  41. Rao N. C., Barsky S. H., Terranova V. P., Liotta L. A. Isolation of a tumor cell laminin receptor. Biochem Biophys Res Commun. 1983 Mar 29;111(3):804–808. doi: 10.1016/0006-291x(83)91370-0. [DOI] [PubMed] [Google Scholar]
  42. Relman D., Tuomanen E., Falkow S., Golenbock D. T., Saukkonen K., Wright S. D. Recognition of a bacterial adhesion by an integrin: macrophage CR3 (alpha M beta 2, CD11b/CD18) binds filamentous hemagglutinin of Bordetella pertussis. Cell. 1990 Jun 29;61(7):1375–1382. doi: 10.1016/0092-8674(90)90701-f. [DOI] [PubMed] [Google Scholar]
  43. Rimoldi M. T., Tenner A. J., Bobak D. A., Joiner K. A. Complement component C1q enhances invasion of human mononuclear phagocytes and fibroblasts by Trypanosoma cruzi trypomastigotes. J Clin Invest. 1989 Dec;84(6):1982–1989. doi: 10.1172/JCI114388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Ruoslahti E., Pierschbacher M. D. New perspectives in cell adhesion: RGD and integrins. Science. 1987 Oct 23;238(4826):491–497. doi: 10.1126/science.2821619. [DOI] [PubMed] [Google Scholar]
  45. Russell D. G., Talamas-Rohana P. Leishmania and the macrophage: a marriage of inconvenience. Immunol Today. 1989 Oct;10(10):328–333. doi: 10.1016/0167-5699(89)90188-6. [DOI] [PubMed] [Google Scholar]
  46. Simpson W. A., Courtney H. S., Ofek I. Interactions of fibronectin with streptococci: the role of fibronectin as a receptor for Streptococcus pyogenes. Rev Infect Dis. 1987 Jul-Aug;9 (Suppl 4):S351–S359. doi: 10.1093/clinids/9.supplement_4.s351. [DOI] [PubMed] [Google Scholar]
  47. Skubitz A. P., McCarthy J. B., Charonis A. S., Furcht L. T. Localization of three distinct heparin-binding domains of laminin by monoclonal antibodies. J Biol Chem. 1988 Apr 5;263(10):4861–4868. [PubMed] [Google Scholar]
  48. Sonnenberg A., Linders C. J., Modderman P. W., Damsky C. H., Aumailley M., Timpl R. Integrin recognition of different cell-binding fragments of laminin (P1, E3, E8) and evidence that alpha 6 beta 1 but not alpha 6 beta 4 functions as a major receptor for fragment E8. J Cell Biol. 1990 Jun;110(6):2145–2155. doi: 10.1083/jcb.110.6.2145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Sonnenberg A., Modderman P. W., Hogervorst F. Laminin receptor on platelets is the integrin VLA-6. Nature. 1988 Dec 1;336(6198):487–489. doi: 10.1038/336487a0. [DOI] [PubMed] [Google Scholar]
  50. Switalski L. M., Speziale P., Hök M., Wadström T., Timpl R. Binding of Streptococcus pyogenes to laminin. J Biol Chem. 1984 Mar 25;259(6):3734–3738. [PubMed] [Google Scholar]
  51. Talamás-Rohana P., Meza I. Interaction between pathogenic amebas and fibronectin: substrate degradation and changes in cytoskeleton organization. J Cell Biol. 1988 May;106(5):1787–1794. doi: 10.1083/jcb.106.5.1787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Tashiro K., Sephel G. C., Weeks B., Sasaki M., Martin G. R., Kleinman H. K., Yamada Y. A synthetic peptide containing the IKVAV sequence from the A chain of laminin mediates cell attachment, migration, and neurite outgrowth. J Biol Chem. 1989 Sep 25;264(27):16174–16182. [PubMed] [Google Scholar]
  53. Timpl R., Rohde H., Robey P. G., Rennard S. I., Foidart J. M., Martin G. R. Laminin--a glycoprotein from basement membranes. J Biol Chem. 1979 Oct 10;254(19):9933–9937. [PubMed] [Google Scholar]
  54. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Turner D. C., Flier L. A., Carbonetto S. Identification of a cell-surface protein involved in PC12 cell-substratum adhesion and neurite outgrowth on laminin and collagen. J Neurosci. 1989 Sep;9(9):3287–3296. doi: 10.1523/JNEUROSCI.09-09-03287.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Velge P., Ouaissi M. A., Cornette J., Afchain D., Capron A. Identification and isolation of Trypanosoma cruzi trypomastigote collagen-binding proteins: possible role in cell-parasite interaction. Parasitology. 1988 Oct;97(Pt 2):255–268. doi: 10.1017/s0031182000058467. [DOI] [PubMed] [Google Scholar]
  57. Wayner E. A., Carter W. G., Piotrowicz R. S., Kunicki T. J. The function of multiple extracellular matrix receptors in mediating cell adhesion to extracellular matrix: preparation of monoclonal antibodies to the fibronectin receptor that specifically inhibit cell adhesion to fibronectin and react with platelet glycoproteins Ic-IIa. J Cell Biol. 1988 Nov;107(5):1881–1891. doi: 10.1083/jcb.107.5.1881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Werk R. How does Toxoplasma gondii enter host cells? Rev Infect Dis. 1985 Jul-Aug;7(4):449–457. doi: 10.1093/clinids/7.4.449. [DOI] [PubMed] [Google Scholar]
  59. Wewer U. M., Liotta L. A., Jaye M., Ricca G. A., Drohan W. N., Claysmith A. P., Rao C. N., Wirth P., Coligan J. E., Albrechtsen R. Altered levels of laminin receptor mRNA in various human carcinoma cells that have different abilities to bind laminin. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7137–7141. doi: 10.1073/pnas.83.19.7137. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES