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. 1995 Aug 1;182(2):345–355. doi: 10.1084/jem.182.2.345

The alpha 4 integrin chain is a ligand for alpha 4 beta 7 and alpha 4 beta 1

PMCID: PMC2192118  PMID: 7629498

Abstract

The heterodimeric alpha 4 integrins alpha 4 beta 7 lymphocyte Peyer's patch adhesion molecule ([LPAM]-1) and alpha 4 beta 1 (very late antigen-4) are cell surface adhesion molecules involved in lymphocyte trafficking and lymphocyte-cell and matrix interactions. Known cellular ligands include vascular cell adhesion molecule (VCAM)-1, which binds to alpha 4 beta 1 and alpha 4 beta 7, and the mucosal addressin cell adhesion molecule (MAdCAM)-1, which binds to alpha 4 beta 7. Here we show that the alpha 4 chain of these integrins can itself serve as a ligand. The alpha 4 chain, immunoaffinity purified and immobilized on glass slides, binds thymocytes and T lymphocytes. Binding exhibits divalent cation requirements and temperature sensitivity which are characteristic of integrin-mediated interactions, and is specifically inhibited by anti-alpha 4 integrin antibodies, which exert their effect at the cell surface. Cells expressing exclusively alpha 4 beta 7 (TK-1) or alpha 4 beta 1 (L1-2) both bound avidly, whereas alpha 4-negative cells did not. A soluble 34-kD alpha 4 chain fragment retained binding activity, and it inhibited lymphocyte adhesion to alpha 4 ligands. It has been shown that alpha 4 integrin binding to fibronectin involves an leucine-aspartic acid-valine (LDV) motif in the HepII/IIICS region of fibronectin (CS-1 peptide), and homologous sequences are important in binding to VCAM-1 and MAdCAM-1. Three conserved LDV motifs occur in the extracellular sequence of alpha 4. A synthetic LDV-containing alpha 4- derived oligopeptide supports alpha 4-integrin-dependent lymphocyte adhesion and blocks binding to the 34-kD alpha 4 chain fragment. Our results suggest that alpha 4 beta 7 and alpha 4 beta 1 integrins may be able to bind to the alpha 4 subunit on adjacent cells, providing a novel mechanism for alpha 4 integrin-mediated and activation-regulated lymphocyte interactions during immune responses.

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Selected References

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  1. Altevogt P., Kohl U., Von Hoegen P., Lang E., Schirrmacher V. Antibody 12-15 cross-reacts with mouse Fc gamma receptors and CD2: study of thymus expression, genetic polymorphism and biosynthesis of the CD2 protein. Eur J Immunol. 1989 Feb;19(2):341–346. doi: 10.1002/eji.1830190219. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bednarczyk J. L., Wygant J. N., Szabo M. C., Molinari-Storey L., Renz M., Fong S., McIntyre B. W. Homotypic leukocyte aggregation triggered by a monoclonal antibody specific for a novel epitope expressed by the integrin beta 1 subunit: conversion of nonresponsive cells by transfecting human integrin alpha 4 subunit cDNA. J Cell Biochem. 1993 Apr;51(4):465–478. doi: 10.1002/jcb.2400510412. [DOI] [PubMed] [Google Scholar]
  4. Berlin C., Berg E. L., Briskin M. J., Andrew D. P., Kilshaw P. J., Holzmann B., Weissman I. L., Hamann A., Butcher E. C. Alpha 4 beta 7 integrin mediates lymphocyte binding to the mucosal vascular addressin MAdCAM-1. Cell. 1993 Jul 16;74(1):185–195. doi: 10.1016/0092-8674(93)90305-a. [DOI] [PubMed] [Google Scholar]
  5. Briskin M. J., McEvoy L. M., Butcher E. C. MAdCAM-1 has homology to immunoglobulin and mucin-like adhesion receptors and to IgA1. Nature. 1993 Jun 3;363(6428):461–464. doi: 10.1038/363461a0. [DOI] [PubMed] [Google Scholar]
  6. Butcher E. C., Scollay R. G., Weissman I. L. Organ specificity of lymphocyte migration: mediation by highly selective lymphocyte interaction with organ-specific determinants on high endothelial venules. Eur J Immunol. 1980 Jul;10(7):556–561. doi: 10.1002/eji.1830100713. [DOI] [PubMed] [Google Scholar]
  7. Campanero M. R., Pulido R., Ursa M. A., Rodríguez-Moya M., de Landázuri M. O., Sánchez-Madrid F. An alternative leukocyte homotypic adhesion mechanism, LFA-1/ICAM-1-independent, triggered through the human VLA-4 integrin. J Cell Biol. 1990 Jun;110(6):2157–2165. doi: 10.1083/jcb.110.6.2157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clements J. M., Newham P., Shepherd M., Gilbert R., Dudgeon T. J., Needham L. A., Edwards R. M., Berry L., Brass A., Humphries M. J. Identification of a key integrin-binding sequence in VCAM-1 homologous to the LDV active site in fibronectin. J Cell Sci. 1994 Aug;107(Pt 8):2127–2135. doi: 10.1242/jcs.107.8.2127. [DOI] [PubMed] [Google Scholar]
  9. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  10. Coffman R. L., Weissman I. L. A monoclonal antibody that recognizes B cells and B cell precursors in mice. J Exp Med. 1981 Feb 1;153(2):269–279. doi: 10.1084/jem.153.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cybulsky M. I., Fries J. W., Williams A. J., Sultan P., Eddy R., Byers M., Shows T., Gimbrone M. A., Jr, Collins T. Gene structure, chromosomal location, and basis for alternative mRNA splicing of the human VCAM1 gene. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7859–7863. doi: 10.1073/pnas.88.17.7859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Ennis E., Isberg R. R., Shimizu Y. Very late antigen 4-dependent adhesion and costimulation of resting human T cells by the bacterial beta 1 integrin ligand invasin. J Exp Med. 1993 Jan 1;177(1):207–212. doi: 10.1084/jem.177.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Garcia-Pardo A., Wayner E. A., Carter W. G., Ferreira O. C., Jr Human B lymphocytes define an alternative mechanism of adhesion to fibronectin. The interaction of the alpha 4 beta 1 integrin with the LHGPEILDVPST sequence of the type III connecting segment is sufficient to promote cell attachment. J Immunol. 1990 May 1;144(9):3361–3366. [PubMed] [Google Scholar]
  15. Groux H., Huet S., Valentin H., Pham D., Bernard A. Suppressor effects and cyclic AMP accumulation by the CD29 molecule of CD4+ lymphocytes. Nature. 1989 May 11;339(6220):152–154. doi: 10.1038/339152a0. [DOI] [PubMed] [Google Scholar]
  16. Guan J. L., Hynes R. O. Lymphoid cells recognize an alternatively spliced segment of fibronectin via the integrin receptor alpha 4 beta 1. Cell. 1990 Jan 12;60(1):53–61. doi: 10.1016/0092-8674(90)90715-q. [DOI] [PubMed] [Google Scholar]
  17. Hahne M., Lenter M., Jäger U., Vestweber D. A novel soluble form of mouse VCAM-1 is generated from a glycolipid-anchored splicing variant. Eur J Immunol. 1994 Feb;24(2):421–428. doi: 10.1002/eji.1830240223. [DOI] [PubMed] [Google Scholar]
  18. Hemler M. E., Huang C., Takada Y., Schwarz L., Strominger J. L., Clabby M. L. Characterization of the cell surface heterodimer VLA-4 and related peptides. J Biol Chem. 1987 Aug 25;262(24):11478–11485. [PubMed] [Google Scholar]
  19. Hession C., Moy P., Tizard R., Chisholm P., Williams C., Wysk M., Burkly L., Miyake K., Kincade P., Lobb R. Cloning of murine and rat vascular cell adhesion molecule-1. Biochem Biophys Res Commun. 1992 Feb 28;183(1):163–169. doi: 10.1016/0006-291x(92)91623-x. [DOI] [PubMed] [Google Scholar]
  20. Holzmann B., McIntyre B. W., Weissman I. L. Identification of a murine Peyer's patch--specific lymphocyte homing receptor as an integrin molecule with an alpha chain homologous to human VLA-4 alpha. Cell. 1989 Jan 13;56(1):37–46. doi: 10.1016/0092-8674(89)90981-1. [DOI] [PubMed] [Google Scholar]
  21. Holzmann B., Weissman I. L. Peyer's patch-specific lymphocyte homing receptors consist of a VLA-4-like alpha chain associated with either of two integrin beta chains, one of which is novel. EMBO J. 1989 Jun;8(6):1735–1741. doi: 10.1002/j.1460-2075.1989.tb03566.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hu M. C., Crowe D. T., Weissman I. L., Holzmann B. Cloning and expression of mouse integrin beta p(beta 7): a functional role in Peyer's patch-specific lymphocyte homing. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8254–8258. doi: 10.1073/pnas.89.17.8254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hubbe M., Kowitz A., Schirrmacher V., Schachner M., Altevogt P. L1 adhesion molecule on mouse leukocytes: regulation and involvement in endothelial cell binding. Eur J Immunol. 1993 Nov;23(11):2927–2931. doi: 10.1002/eji.1830231130. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Kadmon G., Eckert M., Sammar M., Schachner M., Altevogt P. Nectadrin, the heat-stable antigen, is a cell adhesion molecule. J Cell Biol. 1992 Sep;118(5):1245–1258. doi: 10.1083/jcb.118.5.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Komoriya A., Green L. J., Mervic M., Yamada S. S., Yamada K. M., Humphries M. J. The minimal essential sequence for a major cell type-specific adhesion site (CS1) within the alternatively spliced type III connecting segment domain of fibronectin is leucine-aspartic acid-valine. J Biol Chem. 1991 Aug 15;266(23):15075–15079. [PubMed] [Google Scholar]
  27. Lang E., Kohl U., Schirrmacher V., Brossmer R., Altevogt P. Structural basis for altered soybean agglutinin lectin binding between a murine metastatic lymphoma and an adhesive low malignant variant. Exp Cell Res. 1987 Nov;173(1):232–243. doi: 10.1016/0014-4827(87)90348-x. [DOI] [PubMed] [Google Scholar]
  28. Lenter M., Uhlig H., Hamann A., Jenö P., Imhof B., Vestweber D. A monoclonal antibody against an activation epitope on mouse integrin chain beta 1 blocks adhesion of lymphocytes to the endothelial integrin alpha 6 beta 1. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):9051–9055. doi: 10.1073/pnas.90.19.9051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Maeda T., Oyama R., Titani K., Sekiguchi K. Engineering of artificial cell-adhesive proteins by grafting EILDVPST sequence derived from fibronectin. J Biochem. 1993 Jan;113(1):29–35. doi: 10.1093/oxfordjournals.jbchem.a123998. [DOI] [PubMed] [Google Scholar]
  30. Makarem R., Newham P., Askari J. A., Green L. J., Clements J., Edwards M., Humphries M. J., Mould A. P. Competitive binding of vascular cell adhesion molecule-1 and the HepII/IIICS domain of fibronectin to the integrin alpha 4 beta 1. J Biol Chem. 1994 Feb 11;269(6):4005–4011. [PubMed] [Google Scholar]
  31. Miyake K., Medina K., Ishihara K., Kimoto M., Auerbach R., Kincade P. W. A VCAM-like adhesion molecule on murine bone marrow stromal cells mediates binding of lymphocyte precursors in culture. J Cell Biol. 1991 Aug;114(3):557–565. doi: 10.1083/jcb.114.3.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Miyake K., Weissman I. L., Greenberger J. S., Kincade P. W. Evidence for a role of the integrin VLA-4 in lympho-hemopoiesis. J Exp Med. 1991 Mar 1;173(3):599–607. doi: 10.1084/jem.173.3.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mould A. P., Wheldon L. A., Komoriya A., Wayner E. A., Yamada K. M., Humphries M. J. Affinity chromatographic isolation of the melanoma adhesion receptor for the IIICS region of fibronectin and its identification as the integrin alpha 4 beta 1. J Biol Chem. 1990 Mar 5;265(7):4020–4024. [PubMed] [Google Scholar]
  34. Osborn L., Hession C., Tizard R., Vassallo C., Luhowskyj S., Chi-Rosso G., Lobb R. Direct expression cloning of vascular cell adhesion molecule 1, a cytokine-induced endothelial protein that binds to lymphocytes. Cell. 1989 Dec 22;59(6):1203–1211. doi: 10.1016/0092-8674(89)90775-7. [DOI] [PubMed] [Google Scholar]
  35. Osborn L., Vassallo C., Browning B. G., Tizard R., Haskard D. O., Benjamin C. D., Dougas I., Kirchhausen T. Arrangement of domains, and amino acid residues required for binding of vascular cell adhesion molecule-1 to its counter-receptor VLA-4 (alpha 4 beta 1). J Cell Biol. 1994 Feb;124(4):601–608. doi: 10.1083/jcb.124.4.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Polte T., Newman W., Gopal T. V. Full length vascular cell adhesion molecule 1 (VCAM-1). Nucleic Acids Res. 1990 Oct 11;18(19):5901–5901. doi: 10.1093/nar/18.19.5901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Prieto J., Takei F., Gendelman R., Christenson B., Biberfeld P., Patarroyo M. MALA-2, mouse homologue of human adhesion molecule ICAM-1 (CD54). Eur J Immunol. 1989 Sep;19(9):1551–1557. doi: 10.1002/eji.1830190906. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Qian F., Vaux D. L., Weissman I. L. Expression of the integrin alpha 4 beta 1 on melanoma cells can inhibit the invasive stage of metastasis formation. Cell. 1994 May 6;77(3):335–347. doi: 10.1016/0092-8674(94)90149-x. [DOI] [PubMed] [Google Scholar]
  40. Reske-Kunz A. B., Rüde E. Insulin-specific T cell hybridomas derived from (H-2b x H-2k)F1 mice preferably employ F1-unique restriction elements for antigen recognition. Eur J Immunol. 1985 Oct;15(10):1048–1054. doi: 10.1002/eji.1830151017. [DOI] [PubMed] [Google Scholar]
  41. Rice G. E., Munro J. M., Bevilacqua M. P. Inducible cell adhesion molecule 110 (INCAM-110) is an endothelial receptor for lymphocytes. A CD11/CD18-independent adhesion mechanism. J Exp Med. 1990 Apr 1;171(4):1369–1374. doi: 10.1084/jem.171.4.1369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rüegg C., Postigo A. A., Sikorski E. E., Butcher E. C., Pytela R., Erle D. J. Role of integrin alpha 4 beta 7/alpha 4 beta P in lymphocyte adherence to fibronectin and VCAM-1 and in homotypic cell clustering. J Cell Biol. 1992 Apr;117(1):179–189. doi: 10.1083/jcb.117.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sarmiento M., Dialynas D. P., Lancki D. W., Wall K. A., Lorber M. I., Loken M. R., Fitch F. W. Cloned T lymphocytes and monoclonal antibodies as probes for cell surface molecules active in T cell-mediated cytolysis. Immunol Rev. 1982;68:135–169. doi: 10.1111/j.1600-065x.1982.tb01063.x. [DOI] [PubMed] [Google Scholar]
  44. Schirrmacher V., Fogel M., Russmann E., Bosslet K., Altevogt P., Beck L. Antigenic variation in cancer metastasis: immune escape versus immune control. Cancer Metastasis Rev. 1982;1(3):241–274. doi: 10.1007/BF00046830. [DOI] [PubMed] [Google Scholar]
  45. Shimizu Y., Newman W., Tanaka Y., Shaw S. Lymphocyte interactions with endothelial cells. Immunol Today. 1992 Mar;13(3):106–112. doi: 10.1016/0167-5699(92)90151-V. [DOI] [PubMed] [Google Scholar]
  46. Springer T. A. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994 Jan 28;76(2):301–314. doi: 10.1016/0092-8674(94)90337-9. [DOI] [PubMed] [Google Scholar]
  47. Sriramarao P., Steffner P., Gehlsen K. R. Biochemical evidence for a homophilic interaction of the alpha 3 beta 1 integrin. J Biol Chem. 1993 Oct 15;268(29):22036–22041. [PubMed] [Google Scholar]
  48. Strauch U. G., Lifka A., Gosslar U., Kilshaw P. J., Clements J., Holzmann B. Distinct binding specificities of integrins alpha 4 beta 7 (LPAM-1), alpha 4 beta 1 (VLA-4), and alpha IEL beta 7. Int Immunol. 1994 Feb;6(2):263–275. doi: 10.1093/intimm/6.2.263. [DOI] [PubMed] [Google Scholar]
  49. Streeter P. R., Berg E. L., Rouse B. T., Bargatze R. F., Butcher E. C. A tissue-specific endothelial cell molecule involved in lymphocyte homing. Nature. 1988 Jan 7;331(6151):41–46. doi: 10.1038/331041a0. [DOI] [PubMed] [Google Scholar]
  50. Sánchez-Mateos P., Campanero M. R., Balboa M. A., Sánchez-Madrid F. Co-clustering of beta 1 integrins, cytoskeletal proteins, and tyrosine-phosphorylated substrates during integrin-mediated leukocyte aggregation. J Immunol. 1993 Oct 1;151(7):3817–3828. [PubMed] [Google Scholar]
  51. Takada Y., Elices M. J., Crouse C., Hemler M. E. The primary structure of the alpha 4 subunit of VLA-4: homology to other integrins and a possible cell-cell adhesion function. EMBO J. 1989 May;8(5):1361–1368. doi: 10.1002/j.1460-2075.1989.tb03516.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Teixidó J., Parker C. M., Kassner P. D., Hemler M. E. Functional and structural analysis of VLA-4 integrin alpha 4 subunit cleavage. J Biol Chem. 1992 Jan 25;267(3):1786–1791. [PubMed] [Google Scholar]
  53. Vonderheide R. H., Tedder T. F., Springer T. A., Staunton D. E. Residues within a conserved amino acid motif of domains 1 and 4 of VCAM-1 are required for binding to VLA-4. J Cell Biol. 1994 Apr;125(1):215–222. doi: 10.1083/jcb.125.1.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Wadsworth S., Halvorson M. J., Chang A. C., Coligan J. E. Multiple changes in VLA protein glycosylation, expression, and function occur during mouse T cell ontogeny. J Immunol. 1993 Feb 1;150(3):847–857. [PubMed] [Google Scholar]
  55. Wayner E. A., Garcia-Pardo A., Humphries M. J., McDonald J. A., Carter W. G. Identification and characterization of the T lymphocyte adhesion receptor for an alternative cell attachment domain (CS-1) in plasma fibronectin. J Cell Biol. 1989 Sep;109(3):1321–1330. doi: 10.1083/jcb.109.3.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wayner E. A., Kovach N. L. Activation-dependent recognition by hematopoietic cells of the LDV sequence in the V region of fibronectin. J Cell Biol. 1992 Jan;116(2):489–497. doi: 10.1083/jcb.116.2.489. [DOI] [PMC free article] [PubMed] [Google Scholar]

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