Abstract
Activated platelets express P-selectin and release leukocyte chemoattractants; however, they have not been known to express integrin ligands important in the stabilization of leukocyte interactions with the vasculature. We now demonstrate the presence of intercellular adhesion molecular-2 (ICAM-2) (CD102), and lack of expression of other beta 2-integrin ligands, ICAM-1 (CD54) and ICAM-3 (CD50), on the surface of resting and stimulated platelets. ICAM-2 isolated from platelets migrates as a band of 59,000 M(r) in reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Staining of bone marrow aspirates with anti-ICAM-2 mAb demonstrates strong reactivity to megakaryocytes. Using frozen thin sections and immunogold labeling, the antigen was shown to be present on the plasma membrane and surface-connected canalicular system of resting platelets. The average number of ICAM-2 molecules per platelet is 3,000 +/- 230 and does not change after activation. In adhesion assays, resting and stimulated platelets were capable of binding through ICAM-2 to purified leukocyte function-associated antigen-1. Activation of T lymphocytes with PMA stimulated binding to platelets that was Mg2+ dependent and could be specifically inhibited by mAbs to either ICAM-2 or leukocyte function-associated antigen-1. ICAM-2 is the only known beta 2-integrin ligand present on platelets, suggesting that it may play an important role in leukocyte-platelet interactions in inflammation and thrombosis.
Full text
PDF![1243](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3c5/295209/5deb2930e159/jcinvest00021-0341.png)
![1244](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3c5/295209/ab692336d4c3/jcinvest00021-0342.png)
![1245](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3c5/295209/4d7f1539cae2/jcinvest00021-0343.png)
![1246](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3c5/295209/8e006fcd4959/jcinvest00021-0344.png)
![1247](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3c5/295209/1388f2afc672/jcinvest00021-0345.png)
![1248](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3c5/295209/fd8056d00b76/jcinvest00021-0346.png)
![1249](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3c5/295209/64f6f9e7b452/jcinvest00021-0347.png)
![1250](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3c5/295209/8e5c22ff587d/jcinvest00021-0348.png)
![1251](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b3c5/295209/2edd1863ddc7/jcinvest00021-0349.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abts H., Emmerich M., Miltenyi S., Radbruch A., Tesch H. CD20 positive human B lymphocytes separated with the magnetic cell sorter (MACS) can be induced to proliferation and antibody secretion in vitro. J Immunol Methods. 1989 Dec 20;125(1-2):19–28. doi: 10.1016/0022-1759(89)90073-2. [DOI] [PubMed] [Google Scholar]
- Bainton D. F., Miller L. J., Kishimoto T. K., Springer T. A. Leukocyte adhesion receptors are stored in peroxidase-negative granules of human neutrophils. J Exp Med. 1987 Dec 1;166(6):1641–1653. doi: 10.1084/jem.166.6.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bednar M., Smith B., Pinto A., Mullane K. M. Neutrophil depletion suppresses 111In-labeled platelet accumulation in infarcted myocardium. J Cardiovasc Pharmacol. 1985 Sep-Oct;7(5):906–912. doi: 10.1097/00005344-198509000-00014. [DOI] [PubMed] [Google Scholar]
- Buttrum S. M., Hatton R., Nash G. B. Selectin-mediated rolling of neutrophils on immobilized platelets. Blood. 1993 Aug 15;82(4):1165–1174. [PubMed] [Google Scholar]
- Coller B. S. A new murine monoclonal antibody reports an activation-dependent change in the conformation and/or microenvironment of the platelet glycoprotein IIb/IIIa complex. J Clin Invest. 1985 Jul;76(1):101–108. doi: 10.1172/JCI111931. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dustin M. L., Springer T. A. T-cell receptor cross-linking transiently stimulates adhesiveness through LFA-1. Nature. 1989 Oct 19;341(6243):619–624. doi: 10.1038/341619a0. [DOI] [PubMed] [Google Scholar]
- Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
- Ginsberg M. H., Loftus J. C., Plow E. F. Cytoadhesins, integrins, and platelets. Thromb Haemost. 1988 Feb 25;59(1):1–6. [PubMed] [Google Scholar]
- Griffin J. D., Ritz J., Nadler L. M., Schlossman S. F. Expression of myeloid differentiation antigens on normal and malignant myeloid cells. J Clin Invest. 1981 Oct;68(4):932–941. doi: 10.1172/JCI110348. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Griffiths G., McDowall A., Back R., Dubochet J. On the preparation of cryosections for immunocytochemistry. J Ultrastruct Res. 1984 Oct;89(1):65–78. doi: 10.1016/s0022-5320(84)80024-6. [DOI] [PubMed] [Google Scholar]
- Hsu-Lin S., Berman C. L., Furie B. C., August D., Furie B. A platelet membrane protein expressed during platelet activation and secretion. Studies using a monoclonal antibody specific for thrombin-activated platelets. J Biol Chem. 1984 Jul 25;259(14):9121–9126. [PubMed] [Google Scholar]
- Hymes K., Nardi M., Leaf A., Karpatkin S. Role of leuCAM integrins and complement in platelet-monocyte rosette formation induced by immune complexes of human immunodeficiency virus-type 1-immune thrombocytopenic purpura patients. Blood. 1993 May 1;81(9):2375–2380. [PubMed] [Google Scholar]
- Hynes R. O. The complexity of platelet adhesion to extracellular matrices. Thromb Haemost. 1991 Jul 12;66(1):40–43. [PubMed] [Google Scholar]
- Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
- Kieffer N., Phillips D. R. Platelet membrane glycoproteins: functions in cellular interactions. Annu Rev Cell Biol. 1990;6:329–357. doi: 10.1146/annurev.cb.06.110190.001553. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
- Lawrence M. B., Springer T. A. Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion through integrins. Cell. 1991 May 31;65(5):859–873. doi: 10.1016/0092-8674(91)90393-d. [DOI] [PubMed] [Google Scholar]
- Maclouf J. A., Murphy R. C. Transcellular metabolism of neutrophil-derived leukotriene A4 by human platelets. A potential cellular source of leukotriene C4. J Biol Chem. 1988 Jan 5;263(1):174–181. [PubMed] [Google Scholar]
- Marlin S. D., Springer T. A. Purified intercellular adhesion molecule-1 (ICAM-1) is a ligand for lymphocyte function-associated antigen 1 (LFA-1). Cell. 1987 Dec 4;51(5):813–819. doi: 10.1016/0092-8674(87)90104-8. [DOI] [PubMed] [Google Scholar]
- Mayadas T. N., Johnson R. C., Rayburn H., Hynes R. O., Wagner D. D. Leukocyte rolling and extravasation are severely compromised in P selectin-deficient mice. Cell. 1993 Aug 13;74(3):541–554. doi: 10.1016/0092-8674(93)80055-j. [DOI] [PubMed] [Google Scholar]
- McEver R. P. GMP-140: a receptor for neutrophils and monocytes on activated platelets and endothelium. J Cell Biochem. 1991 Feb;45(2):156–161. doi: 10.1002/jcb.240450206. [DOI] [PubMed] [Google Scholar]
- McEver R. P., Martin M. N. A monoclonal antibody to a membrane glycoprotein binds only to activated platelets. J Biol Chem. 1984 Aug 10;259(15):9799–9804. [PubMed] [Google Scholar]
- McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
- Miller L. J., Bainton D. F., Borregaard N., Springer T. A. Stimulated mobilization of monocyte Mac-1 and p150,95 adhesion proteins from an intracellular vesicular compartment to the cell surface. J Clin Invest. 1987 Aug;80(2):535–544. doi: 10.1172/JCI113102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller M. D., Krangel M. S. Biology and biochemistry of the chemokines: a family of chemotactic and inflammatory cytokines. Crit Rev Immunol. 1992;12(1-2):17–46. [PubMed] [Google Scholar]
- Miltenyi S., Müller W., Weichel W., Radbruch A. High gradient magnetic cell separation with MACS. Cytometry. 1990;11(2):231–238. doi: 10.1002/cyto.990110203. [DOI] [PubMed] [Google Scholar]
- Nagata K., Tsuji T., Todoroki N., Katagiri Y., Tanoue K., Yamazaki H., Hanai N., Irimura T. Activated platelets induce superoxide anion release by monocytes and neutrophils through P-selectin (CD62). J Immunol. 1993 Sep 15;151(6):3267–3273. [PubMed] [Google Scholar]
- Nesheim M. E., Pittman D. D., Wang J. H., Slonosky D., Giles A. R., Kaufman R. J. The binding of 35S-labeled recombinant factor VIII to activated and unactivated human platelets. J Biol Chem. 1988 Nov 5;263(31):16467–16470. [PubMed] [Google Scholar]
- Palabrica T., Lobb R., Furie B. C., Aronovitz M., Benjamin C., Hsu Y. M., Sajer S. A., Furie B. Leukocyte accumulation promoting fibrin deposition is mediated in vivo by P-selectin on adherent platelets. Nature. 1992 Oct 29;359(6398):848–851. doi: 10.1038/359848a0. [DOI] [PubMed] [Google Scholar]
- Reinherz E. L., Schlossman S. F. The differentiation and function of human T lymphocytes. Cell. 1980 Apr;19(4):821–827. doi: 10.1016/0092-8674(80)90072-0. [DOI] [PubMed] [Google Scholar]
- Rothlein R., Dustin M. L., Marlin S. D., Springer T. A. A human intercellular adhesion molecule (ICAM-1) distinct from LFA-1. J Immunol. 1986 Aug 15;137(4):1270–1274. [PubMed] [Google Scholar]
- 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]
- Smyth S. S., Joneckis C. C., Parise L. V. Regulation of vascular integrins. Blood. 1993 Jun 1;81(11):2827–2843. [PubMed] [Google Scholar]
- Springer T. A. Adhesion receptors of the immune system. Nature. 1990 Aug 2;346(6283):425–434. doi: 10.1038/346425a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Staunton D. E., Dustin M. L., Springer T. A. Functional cloning of ICAM-2, a cell adhesion ligand for LFA-1 homologous to ICAM-1. Nature. 1989 May 4;339(6219):61–64. doi: 10.1038/339061a0. [DOI] [PubMed] [Google Scholar]
- Stenberg P. E., McEver R. P., Shuman M. A., Jacques Y. V., Bainton D. F. A platelet alpha-granule membrane protein (GMP-140) is expressed on the plasma membrane after activation. J Cell Biol. 1985 Sep;101(3):880–886. doi: 10.1083/jcb.101.3.880. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stenberg P. E., Shuman M. A., Levine S. P., Bainton D. F. Optimal techniques for the immunocytochemical demonstration of beta-thromboglobulin, platelet factor 4, and fibrinogen in the alpha granules of unstimulated platelets. Histochem J. 1984 Sep;16(9):983–1001. doi: 10.1007/BF01003853. [DOI] [PubMed] [Google Scholar]
- Timmons S., Hawiger J. Separation of human platelets from plasma proteins including factor VIII VWF by a combined albumin gradient-gel filtration method using HEPES buffer. Thromb Res. 1978 Feb;12(2):297–306. doi: 10.1016/0049-3848(78)90300-6. [DOI] [PubMed] [Google Scholar]
- Tokuyasu K. T. Present state of immunocryoultramicrotomy. J Histochem Cytochem. 1983 Jan;31(1A):164–167. [PubMed] [Google Scholar]
- Tuszynski G. P., Knight L. C., Kornecki E., Srivastava S. Labeling of platelet surface proteins with 125I by the iodogen method. Anal Biochem. 1983 Apr 1;130(1):166–170. doi: 10.1016/0003-2697(83)90664-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Yelton D. E., Desaymard C., Scharff M. D. Use of monoclonal anti-mouse immunoglobulin to detect mouse antibodies. Hybridoma. 1981;1(1):5–11. doi: 10.1089/hyb.1.1981.1.5. [DOI] [PubMed] [Google Scholar]
- de Fougerolles A. R., Springer T. A. Intercellular adhesion molecule 3, a third adhesion counter-receptor for lymphocyte function-associated molecule 1 on resting lymphocytes. J Exp Med. 1992 Jan 1;175(1):185–190. doi: 10.1084/jem.175.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Fougerolles A. R., Stacker S. A., Schwarting R., Springer T. A. Characterization of ICAM-2 and evidence for a third counter-receptor for LFA-1. J Exp Med. 1991 Jul 1;174(1):253–267. doi: 10.1084/jem.174.1.253. [DOI] [PMC free article] [PubMed] [Google Scholar]