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. 1988 Jul 1;168(1):279–292. doi: 10.1084/jem.168.1.279

Complement receptor type 3 (CR3) binds to an Arg-Gly-Asp-containing region of the major surface glycoprotein, gp63, of Leishmania promastigotes

PMCID: PMC2188978  PMID: 3294332

Abstract

The major surface glycoprotein of Leishmania promastigotes, gp63, was isolated and reconstituted into a lipid membrane immobilized on the surface of 5-micron-diameter silica beads. These beads bound to the macrophage (MO), and the extent of binding correlated with the density of gp63 on the bead. The bead thus facilitated analysis of the binding specificity of a single ligand, gp63, without contribution from other molecules present on the surface of intact promastigotes. Plating of MO onto substrates coated with antibodies directed against several cell surface receptors indicated that the complement receptor CR3 was necessary for binding gp63. CR3 recognizes a portion of C3 that contains the sequence R G D. Since gp63 also contains such a sequence, we tested the ability of a synthetic peptide based on the R G D- containing region of gp63 to inhibit the binding of gp63 beads. The R G D-containing peptide from gp63 inhibited the binding of both gp63 beads and EC3bi to MO. Similarly, peptides previously shown to inhibit the binding of C3bi also inhibited the attachment of gp63 beads. The synthetic peptide from the R G D region of gp63 also reduced the binding of intact promastigotes to MO. These results indicate that gp63 binds directly to CR3.

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Selected References

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  1. Alexander J., Russell D. G. Parasite antigens, their role in protection, diagnosis and escape: the leishmaniases. Curr Top Microbiol Immunol. 1985;120:43–67. doi: 10.1007/978-3-662-09197-5_4. [DOI] [PubMed] [Google Scholar]
  2. Alexander J., Vickerman K. Fusion of host cell secondary lysosomes with the parasitophorous vacuoles of Leishmania mexicana-infected macrophages. J Protozool. 1975 Nov;22(4):502–508. doi: 10.1111/j.1550-7408.1975.tb05219.x. [DOI] [PubMed] [Google Scholar]
  3. Barnstable C. J., Bodmer W. F., Brown G., Galfre G., Milstein C., Williams A. F., Ziegler A. Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens-new tools for genetic analysis. Cell. 1978 May;14(1):9–20. doi: 10.1016/0092-8674(78)90296-9. [DOI] [PubMed] [Google Scholar]
  4. Berton G., Gordon S. Modulation of macrophage mannosyl-specific receptors by cultivation on immobilized zymosan. Effects on superoxide-anion release and phagocytosis. Immunology. 1983 Aug;49(4):705–715. [PMC free article] [PubMed] [Google Scholar]
  5. Blackwell J. M., Ezekowitz R. A., Roberts M. B., Channon J. Y., Sim R. B., Gordon S. Macrophage complement and lectin-like receptors bind Leishmania in the absence of serum. J Exp Med. 1985 Jul 1;162(1):324–331. doi: 10.1084/jem.162.1.324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bordier C., Etges R. J., Ward J., Turner M. J., Cardoso de Almeida M. L. Leishmania and Trypanosoma surface glycoproteins have a common glycophospholipid membrane anchor. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5988–5991. doi: 10.1073/pnas.83.16.5988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bouvier J., Etges R. J., Bordier C. Identification and purification of membrane and soluble forms of the major surface protein of Leishmania promastigotes. J Biol Chem. 1985 Dec 15;260(29):15504–15509. [PubMed] [Google Scholar]
  8. Brun R., Schönenberger Cultivation and in vitro cloning or procyclic culture forms of Trypanosoma brucei in a semi-defined medium. Short communication. Acta Trop. 1979 Sep;36(3):289–292. [PubMed] [Google Scholar]
  9. 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]
  10. Bullock W. E., Wright S. D. Role of the adherence-promoting receptors, CR3, LFA-1, and p150,95, in binding of Histoplasma capsulatum by human macrophages. J Exp Med. 1987 Jan 1;165(1):195–210. doi: 10.1084/jem.165.1.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Button L. L., McMaster W. R. Molecular cloning of the major surface antigen of leishmania. J Exp Med. 1988 Feb 1;167(2):724–729. doi: 10.1084/jem.167.2.724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chang C. S., Chang K. P. Monoclonal antibody affinity purification of a Leishmania membrane glycoprotein and its inhibition of leishmania-macrophage binding. Proc Natl Acad Sci U S A. 1986 Jan;83(1):100–104. doi: 10.1073/pnas.83.1.100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chang K. P., Dwyer D. M. Multiplication of a human parasite (Leishmania donovani) in phagolysosomes of hamster macrophages in vitro. Science. 1976 Aug 20;193(4254):678–680. doi: 10.1126/science.948742. [DOI] [PubMed] [Google Scholar]
  14. Chang K. P. Leishmania donovani-macrophage binding mediated by surface glycoproteins/antigens: characterization in vitro by a radioisotopic assay. Mol Biochem Parasitol. 1981 Nov;4(1-2):67–76. doi: 10.1016/0166-6851(81)90030-x. [DOI] [PubMed] [Google Scholar]
  15. Channon J. Y., Roberts M. B., Blackwell J. M. A study of the differential respiratory burst activity elicited by promastigotes and amastigotes of Leishmania donovani in murine resident peritoneal macrophages. Immunology. 1984 Oct;53(2):345–355. [PMC free article] [PubMed] [Google Scholar]
  16. English D., Andersen B. R. Single-step separation of red blood cells. Granulocytes and mononuclear leukocytes on discontinuous density gradients of Ficoll-Hypaque. J Immunol Methods. 1974 Aug;5(3):249–252. doi: 10.1016/0022-1759(74)90109-4. [DOI] [PubMed] [Google Scholar]
  17. Goldstein S. A., Mescher M. F. Cell-sized, supported artificial membranes (pseudocytes): response of precursor cytotoxic T lymphocytes to class I MHC proteins. J Immunol. 1986 Dec 1;137(11):3383–3392. [PMC free article] [PubMed] [Google Scholar]
  18. Handman E., Goding J. W. The Leishmania receptor for macrophages is a lipid-containing glycoconjugate. EMBO J. 1985 Feb;4(2):329–336. doi: 10.1002/j.1460-2075.1985.tb03633.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hemler M. E., Ware C. F., Strominger J. L. Characterization of a novel differentiation antigen complex recognize by a monoclonal antibody (A-1A5): unique activation-specific molecular forms on stimulated T cells. J Immunol. 1983 Jul;131(1):334–340. [PubMed] [Google Scholar]
  20. Hetland G., Eskeland T. Formation of the functional alternative pathway of complement by human monocytes in vitro as demonstrated by phagocytosis of agarose beads. Scand J Immunol. 1986 Mar;23(3):301–308. doi: 10.1111/j.1365-3083.1986.tb01971.x. [DOI] [PubMed] [Google Scholar]
  21. Johnston R. B., Jr, Lehmeyer J. E., Guthrie L. A. Generation of superoxide anion and chemiluminescence by human monocytes during phagocytosis and on contact with surface-bound immunoglobulin G. J Exp Med. 1976 Jun 1;143(6):1551–1556. doi: 10.1084/jem.143.6.1551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Klempner M. S., Cendron M., Wyler D. J. Attachment of plasma membrane vesicles of human macrophages to Leishmania tropica promastigotes. J Infect Dis. 1983 Sep;148(3):377–384. doi: 10.1093/infdis/148.3.377. [DOI] [PubMed] [Google Scholar]
  23. Kweider M., Lemesre J. L., Darcy F., Kusnierz J. P., Capron A., Santoro F. Infectivity of Leishmania braziliensis promastigotes is dependent on the increasing expression of a 65,000-dalton surface antigen. J Immunol. 1987 Jan 1;138(1):299–305. [PubMed] [Google Scholar]
  24. Lam S. C., Plow E. F., Smith M. A., Andrieux A., Ryckwaert J. J., Marguerie G., Ginsberg M. H. Evidence that arginyl-glycyl-aspartate peptides and fibrinogen gamma chain peptides share a common binding site on platelets. J Biol Chem. 1987 Jan 25;262(3):947–950. [PubMed] [Google Scholar]
  25. Lanier L. L., Arnaout M. A., Schwarting R., Warner N. L., Ross G. D. p150/95, Third member of the LFA-1/CR3 polypeptide family identified by anti-Leu M5 monoclonal antibody. Eur J Immunol. 1985 Jul;15(7):713–718. doi: 10.1002/eji.1830150714. [DOI] [PubMed] [Google Scholar]
  26. Mosser D. M., Edelson P. J. The mouse macrophage receptor for C3bi (CR3) is a major mechanism in the phagocytosis of Leishmania promastigotes. J Immunol. 1985 Oct;135(4):2785–2789. [PubMed] [Google Scholar]
  27. Mosser D. M., Edelson P. J. The third component of complement (C3) is responsible for the intracellular survival of Leishmania major. 1987 May 28-Jun 3Nature. 327(6120):329–331. doi: 10.1038/327329b0. [DOI] [PubMed] [Google Scholar]
  28. Mosser D. M., Vlassara H., Edelson P. J., Cerami A. Leishmania promastigotes are recognized by the macrophage receptor for advanced glycosylation endproducts. J Exp Med. 1987 Jan 1;165(1):140–145. doi: 10.1084/jem.165.1.140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Orlandi P. A., Jr, Turco S. J. Structure of the lipid moiety of the Leishmania donovani lipophosphoglycan. J Biol Chem. 1987 Jul 25;262(21):10384–10391. [PubMed] [Google Scholar]
  30. Pierschbacher M. D., Ruoslahti E. Variants of the cell recognition site of fibronectin that retain attachment-promoting activity. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5985–5988. doi: 10.1073/pnas.81.19.5985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rizvi F. S., Ouaissi M. A., Marty B., Santoro F., Capron A. The major surface protein of Leishmania promastigotes is a fibronectin-like molecule. Eur J Immunol. 1988 Mar;18(3):473–476. doi: 10.1002/eji.1830180323. [DOI] [PubMed] [Google Scholar]
  32. Ruoslahti E., Pierschbacher M. D. Arg-Gly-Asp: a versatile cell recognition signal. Cell. 1986 Feb 28;44(4):517–518. doi: 10.1016/0092-8674(86)90259-x. [DOI] [PubMed] [Google Scholar]
  33. Russell D. G., Alexander J. Effective immunization against cutaneous leishmaniasis with defined membrane antigens reconstituted into liposomes. J Immunol. 1988 Feb 15;140(4):1274–1279. [PubMed] [Google Scholar]
  34. Russell D. G. The macrophage-attachment glycoprotein gp63 is the predominant C3-acceptor site on Leishmania mexicana promastigotes. Eur J Biochem. 1987 Apr 1;164(1):213–221. doi: 10.1111/j.1432-1033.1987.tb11013.x. [DOI] [PubMed] [Google Scholar]
  35. Russell D. G., Wilhelm H. The involvement of the major surface glycoprotein (gp63) of Leishmania promastigotes in attachment to macrophages. J Immunol. 1986 Apr 1;136(7):2613–2620. [PubMed] [Google Scholar]
  36. Sacks D. L., Perkins P. V. Identification of an infective stage of Leishmania promastigotes. Science. 1984 Mar 30;223(4643):1417–1419. doi: 10.1126/science.6701528. [DOI] [PubMed] [Google Scholar]
  37. Sacks D. L., da Silva R. P. The generation of infective stage Leishmania major promastigotes is associated with the cell-surface expression and release of a developmentally regulated glycolipid. J Immunol. 1987 Nov 1;139(9):3099–3106. [PubMed] [Google Scholar]
  38. Sanchez-Madrid F., Krensky A. M., Ware C. F., Robbins E., Strominger J. L., Burakoff S. J., Springer T. A. Three distinct antigens associated with human T-lymphocyte-mediated cytolysis: LFA-1, LFA-2, and LFA-3. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7489–7493. doi: 10.1073/pnas.79.23.7489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Turco S. J., Hull S. R., Orlandi P. A., Jr, Shepherd S. D., Homans S. W., Dwek R. A., Rademacher T. W. Structure of the major carbohydrate fragment of the Leishmania donovani lipophosphoglycan. Biochemistry. 1987 Sep 22;26(19):6233–6238. doi: 10.1021/bi00393a042. [DOI] [PubMed] [Google Scholar]
  40. Wozencraft A. O., Sayers G., Blackwell J. M. Macrophage type 3 complement receptors mediate serum-independent binding of Leishmania donovani. Detection of macrophage-derived complement on the parasite surface by immunoelectron microscopy. J Exp Med. 1986 Oct 1;164(4):1332–1337. doi: 10.1084/jem.164.4.1332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wright S. D., Detmers P. A., Jong M. T., Meyer B. C. Interferon-gamma depresses binding of ligand by C3b and C3bi receptors on cultured human monocytes, an effect reversed by fibronectin. J Exp Med. 1986 May 1;163(5):1245–1259. doi: 10.1084/jem.163.5.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wright S. D., Jong M. T. Adhesion-promoting receptors on human macrophages recognize Escherichia coli by binding to lipopolysaccharide. J Exp Med. 1986 Dec 1;164(6):1876–1888. doi: 10.1084/jem.164.6.1876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wright S. D., Meyer B. C. Fibronectin receptor of human macrophages recognizes the sequence Arg-Gly-Asp-Ser. J Exp Med. 1985 Aug 1;162(2):762–767. doi: 10.1084/jem.162.2.762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wright S. D., Rao P. E., Van Voorhis W. C., Craigmyle L. S., Iida K., Talle M. A., Westberg E. F., Goldstein G., Silverstein S. C. Identification of the C3bi receptor of human monocytes and macrophages by using monoclonal antibodies. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5699–5703. doi: 10.1073/pnas.80.18.5699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wright S. D., Reddy P. A., Jong M. T., Erickson B. W. C3bi receptor (complement receptor type 3) recognizes a region of complement protein C3 containing the sequence Arg-Gly-Asp. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1965–1968. doi: 10.1073/pnas.84.7.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wright S. D., Silverstein S. C. Receptors for C3b and C3bi promote phagocytosis but not the release of toxic oxygen from human phagocytes. J Exp Med. 1983 Dec 1;158(6):2016–2023. doi: 10.1084/jem.158.6.2016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wright S. D., Silverstein S. C. Tumor-promoting phorbol esters stimulate C3b and C3b' receptor-mediated phagocytosis in cultured human monocytes. J Exp Med. 1982 Oct 1;156(4):1149–1164. doi: 10.1084/jem.156.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. da Silva R., Sacks D. L. Metacyclogenesis is a major determinant of Leishmania promastigote virulence and attenuation. Infect Immun. 1987 Nov;55(11):2802–2806. doi: 10.1128/iai.55.11.2802-2806.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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