Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1997 Sep 1;100(5):996–1005. doi: 10.1172/JCI119650

Platelet activation In mice and human Helicobacter pylori infection.

J I Elizalde 1, J Gómez 1, J Panés 1, M Lozano 1, M Casadevall 1, J Ramírez 1, P Pizcueta 1, F Marco 1, F D Rojas 1, D N Granger 1, J M Piqué 1
PMCID: PMC508274  PMID: 9276716

Abstract

Extracts of Helicobacter pylori (HP) have been shown to induce leukocyte adhesion in mesenteric venules, but the effects of HP infection on gastric microvessels are unknown. Inflammatory cell interactions in the gastric microcirculation were studied by intravital videomicroscopy in mice inoculated with either saline or fresh isolates of HP. Platelet aggregates were detected and quantified in murine portal blood, while endothelial P-selectin expression was determined using the dual radiolabeled mAb technique. Platelet activation and aggregation were studied in HP-infected patients and controls by measuring the platelet-aggregate ratio and platelet P-selectin expression. HP infection induced a marked increase in the flux of rolling leukocytes and the appearance of platelet and leukocyte- platelet aggregates in murine gastric venules. The HP-induced rolling and platelet aggregate formation was abrogated by mAbs against L- or P-, but not E- selectin. Endothelial cell expression of P-selectin was not altered, but platelet P-selectin expression was enhanced in HP-infected mice. Circulating platelet aggregates and activated platelets were also detected in HP-infected patients. These findings indicate that platelet activation and aggregation contribute to the microvascular dysfunction and inflammatory cell recruitment associated with HP infections.

Full Text

The Full Text of this article is available as a PDF (308.6 KB).

Selected References

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

  1. Auchampach J. A., Oliver M. G., Anderson D. C., Manning A. M. Cloning, sequence comparison and in vivo expression of the gene encoding rat P-selectin. Gene. 1994 Aug 5;145(2):251–255. doi: 10.1016/0378-1119(94)90015-9. [DOI] [PubMed] [Google Scholar]
  2. Austrup F., Vestweber D., Borges E., Löhning M., Bräuer R., Herz U., Renz H., Hallmann R., Scheffold A., Radbruch A. P- and E-selectin mediate recruitment of T-helper-1 but not T-helper-2 cells into inflammed tissues. Nature. 1997 Jan 2;385(6611):81–83. doi: 10.1038/385081a0. [DOI] [PubMed] [Google Scholar]
  3. Bayerdörffer E., Lehn N., Hatz R., Mannes G. A., Oertel H., Sauerbruch T., Stolte M. Difference in expression of Helicobacter pylori gastritis in antrum and body. Gastroenterology. 1992 May;102(5):1575–1582. doi: 10.1016/0016-5085(92)91716-h. [DOI] [PubMed] [Google Scholar]
  4. Bosse R., Vestweber D. Only simultaneous blocking of the L- and P-selectin completely inhibits neutrophil migration into mouse peritoneum. Eur J Immunol. 1994 Dec;24(12):3019–3024. doi: 10.1002/eji.1830241215. [DOI] [PubMed] [Google Scholar]
  5. Carlos T. M., Harlan J. M. Leukocyte-endothelial adhesion molecules. Blood. 1994 Oct 1;84(7):2068–2101. [PubMed] [Google Scholar]
  6. Colli S., Eligini S., Lalli M., Tremoli E. Platelet-neutrophil interaction and superoxide anion generation: involvement of purine nucleotides. Free Radic Biol Med. 1996;20(3):271–278. doi: 10.1016/0891-5849(95)02032-2. [DOI] [PubMed] [Google Scholar]
  7. Cover T. L., Puryear W., Perez-Perez G. I., Blaser M. J. Effect of urease on HeLa cell vacuolation induced by Helicobacter pylori cytotoxin. Infect Immun. 1991 Apr;59(4):1264–1270. doi: 10.1128/iai.59.4.1264-1270.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crowe S. E., Alvarez L., Dytoc M., Hunt R. H., Muller M., Sherman P., Patel J., Jin Y., Ernst P. B. Expression of interleukin 8 and CD54 by human gastric epithelium after Helicobacter pylori infection in vitro. Gastroenterology. 1995 Jan;108(1):65–74. doi: 10.1016/0016-5085(95)90009-8. [DOI] [PubMed] [Google Scholar]
  9. Cutler A. F. Testing for Helicobacter pylori in clinical practice. Am J Med. 1996 May 20;100(5A):35S–41S. doi: 10.1016/s0002-9343(96)80227-0. [DOI] [PubMed] [Google Scholar]
  10. Diacovo T. G., Puri K. D., Warnock R. A., Springer T. A., von Andrian U. H. Platelet-mediated lymphocyte delivery to high endothelial venules. Science. 1996 Jul 12;273(5272):252–255. doi: 10.1126/science.273.5272.252. [DOI] [PubMed] [Google Scholar]
  11. Elstad M. R., La Pine T. R., Cowley F. S., McEver R. P., McIntyre T. M., Prescott S. M., Zimmerman G. A. P-selectin regulates platelet-activating factor synthesis and phagocytosis by monocytes. J Immunol. 1995 Aug 15;155(4):2109–2122. [PubMed] [Google Scholar]
  12. Enders G., Brooks W., von Jan N., Lehn N., Bayerdörffer E., Hatz R. Expression of adhesion molecules on human granulocytes after stimulation with Helicobacter pylori membrane proteins: comparison with membrane proteins from other bacteria. Infect Immun. 1995 Jul;63(7):2473–2477. doi: 10.1128/iai.63.7.2473-2477.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eppihimer M. J., Wolitzky B., Anderson D. C., Labow M. A., Granger D. N. Heterogeneity of expression of E- and P-selectins in vivo. Circ Res. 1996 Sep;79(3):560–569. doi: 10.1161/01.res.79.3.560. [DOI] [PubMed] [Google Scholar]
  14. Evangelista V., Piccardoni P., White J. G., de Gaetano G., Cerletti C. Cathepsin G-dependent platelet stimulation by activated polymorphonuclear leukocytes and its inhibition by antiproteinases: role of P-selectin-mediated cell-cell adhesion. Blood. 1993 Jun 1;81(11):2947–2957. [PubMed] [Google Scholar]
  15. 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]
  16. Frenette P. S., Johnson R. C., Hynes R. O., Wagner D. D. Platelets roll on stimulated endothelium in vivo: an interaction mediated by endothelial P-selectin. Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7450–7454. doi: 10.1073/pnas.92.16.7450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gawaz M., Neumann F. J., Ott I., Schiessler A., Schömig A. Platelet function in acute myocardial infarction treated with direct angioplasty. Circulation. 1996 Jan 15;93(2):229–237. doi: 10.1161/01.cir.93.2.229. [DOI] [PubMed] [Google Scholar]
  18. Ghiara P., Marchetti M., Blaser M. J., Tummuru M. K., Cover T. L., Segal E. D., Tompkins L. S., Rappuoli R. Role of the Helicobacter pylori virulence factors vacuolating cytotoxin, CagA, and urease in a mouse model of disease. Infect Immun. 1995 Oct;63(10):4154–4160. doi: 10.1128/iai.63.10.4154-4160.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Grober J. S., Bowen B. L., Ebling H., Athey B., Thompson C. B., Fox D. A., Stoolman L. M. Monocyte-endothelial adhesion in chronic rheumatoid arthritis. In situ detection of selectin and integrin-dependent interactions. J Clin Invest. 1993 Jun;91(6):2609–2619. doi: 10.1172/JCI116500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Henninger D. D., Panés J., Eppihimer M., Russell J., Gerritsen M., Anderson D. C., Granger D. N. Cytokine-induced VCAM-1 and ICAM-1 expression in different organs of the mouse. J Immunol. 1997 Feb 15;158(4):1825–1832. [PubMed] [Google Scholar]
  21. Janssen G. H., Tangelder G. J., Oude Egbrink M. G., Reneman R. S. Spontaneous leukocyte rolling in venules in untraumatized skin of conscious and anesthetized animals. Am J Physiol. 1994 Sep;267(3 Pt 2):H1199–H1204. doi: 10.1152/ajpheart.1994.267.3.H1199. [DOI] [PubMed] [Google Scholar]
  22. Jones D. A., Abbassi O., McIntire L. V., McEver R. P., Smith C. W. P-selectin mediates neutrophil rolling on histamine-stimulated endothelial cells. Biophys J. 1993 Oct;65(4):1560–1569. doi: 10.1016/S0006-3495(93)81195-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kozol R., Domanowski A., Jaszewski R., Czanko R., McCurdy B., Prasad M., Fromm B., Calzada R. Neutrophil chemotaxis in gastric mucosa. A signal-to-response comparison. Dig Dis Sci. 1991 Sep;36(9):1277–1280. doi: 10.1007/BF01307522. [DOI] [PubMed] [Google Scholar]
  24. Kubes P., Jutila M., Payne D. Therapeutic potential of inhibiting leukocyte rolling in ischemia/reperfusion. J Clin Invest. 1995 Jun;95(6):2510–2519. doi: 10.1172/JCI117952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kurose I., Granger D. N., Evans D. J., Jr, Evans D. G., Graham D. Y., Miyasaka M., Anderson D. C., Wolf R. E., Cepinskas G., Kvietys P. R. Helicobacter pylori-induced microvascular protein leakage in rats: role of neutrophils, mast cells, and platelets. Gastroenterology. 1994 Jul;107(1):70–79. doi: 10.1016/0016-5085(94)90062-0. [DOI] [PubMed] [Google Scholar]
  26. Lalor P., Nash G. B. Adhesion of flowing leucocytes to immobilized platelets. Br J Haematol. 1995 Apr;89(4):725–732. doi: 10.1111/j.1365-2141.1995.tb08408.x. [DOI] [PubMed] [Google Scholar]
  27. Ley K., Bullard D. C., Arbonés M. L., Bosse R., Vestweber D., Tedder T. F., Beaudet A. L. Sequential contribution of L- and P-selectin to leukocyte rolling in vivo. J Exp Med. 1995 Feb 1;181(2):669–675. doi: 10.1084/jem.181.2.669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lorant D. E., McEver R. P., McIntyre T. M., Moore K. L., Prescott S. M., Zimmerman G. A. Activation of polymorphonuclear leukocytes reduces their adhesion to P-selectin and causes redistribution of ligands for P-selectin on their surfaces. J Clin Invest. 1995 Jul;96(1):171–182. doi: 10.1172/JCI118018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ma L., Raycroft L., Asa D., Anderson D. C., Geng J. G. A sialoglycoprotein from human leukocytes functions as a ligand for P-selectin. J Biol Chem. 1994 Nov 4;269(44):27739–27746. [PubMed] [Google Scholar]
  30. Mai U. E., Perez-Perez G. I., Allen J. B., Wahl S. M., Blaser M. J., Smith P. D. Surface proteins from Helicobacter pylori exhibit chemotactic activity for human leukocytes and are present in gastric mucosa. J Exp Med. 1992 Feb 1;175(2):517–525. doi: 10.1084/jem.175.2.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mai U. E., Perez-Perez G. I., Wahl L. M., Wahl S. M., Blaser M. J., Smith P. D. Soluble surface proteins from Helicobacter pylori activate monocytes/macrophages by lipopolysaccharide-independent mechanism. J Clin Invest. 1991 Mar;87(3):894–900. doi: 10.1172/JCI115095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Marchetti M., Aricò B., Burroni D., Figura N., Rappuoli R., Ghiara P. Development of a mouse model of Helicobacter pylori infection that mimics human disease. Science. 1995 Mar 17;267(5204):1655–1658. doi: 10.1126/science.7886456. [DOI] [PubMed] [Google Scholar]
  33. Maugeri N., Evangelista V., Celardo A., Dell'Elba G., Martelli N., Piccardoni P., de Gaetano G., Cerletti C. Polymorphonuclear leukocyte-platelet interaction: role of P-selectin in thromboxane B2 and leukotriene C4 cooperative synthesis. Thromb Haemost. 1994 Sep;72(3):450–456. [PubMed] [Google Scholar]
  34. Mendall M. A., Goggin P. M., Molineaux N., Levy J., Toosy T., Strachan D., Camm A. J., Northfield T. C. Relation of Helicobacter pylori infection and coronary heart disease. Br Heart J. 1994 May;71(5):437–439. doi: 10.1136/hrt.71.5.437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Nolte D., Hecht R., Schmid P., Botzlar A., Menger M. D., Neumueller C., Sinowatz F., Vestweber D., Messmer K. Role of Mac-1 and ICAM-1 in ischemia-reperfusion injury in a microcirculation model of BALB/C mice. Am J Physiol. 1994 Oct;267(4 Pt 2):H1320–H1328. doi: 10.1152/ajpheart.1994.267.4.H1320. [DOI] [PubMed] [Google Scholar]
  37. Panés J., Perry M. A., Anderson D. C., Manning A., Leone B., Cepinskas G., Rosenbloom C. L., Miyasaka M., Kvietys P. R., Granger D. N. Regional differences in constitutive and induced ICAM-1 expression in vivo. Am J Physiol. 1995 Dec;269(6 Pt 2):H1955–H1964. doi: 10.1152/ajpheart.1995.269.6.H1955. [DOI] [PubMed] [Google Scholar]
  38. Parmentier S., McGregor L., Catimel B., Leung L. L., McGregor J. L. Inhibition of platelet functions by a monoclonal antibody (LYP20) directed against a granule membrane glycoprotein (GMP-140/PADGEM). Blood. 1991 Apr 15;77(8):1734–1739. [PubMed] [Google Scholar]
  39. Patel P., Carrington D., Strachan D. P., Leatham E., Goggin P., Northfield T. C., Mendall M. A. Fibrinogen: a link between chronic infection and coronary heart disease. Lancet. 1994 Jun 25;343(8913):1634–1635. doi: 10.1016/s0140-6736(94)93084-8. [DOI] [PubMed] [Google Scholar]
  40. Reed M. W., Miller F. N. Importance of light dose in fluorescent microscopy. Microvasc Res. 1988 Jul;36(1):104–107. doi: 10.1016/0026-2862(88)90042-8. [DOI] [PubMed] [Google Scholar]
  41. Rinder H. M., Bonan J. L., Rinder C. S., Ault K. A., Smith B. R. Dynamics of leukocyte-platelet adhesion in whole blood. Blood. 1991 Oct 1;78(7):1730–1737. [PubMed] [Google Scholar]
  42. Scharf R. E., Tomer A., Marzec U. M., Teirstein P. S., Ruggeri Z. M., Harker L. A. Activation of platelets in blood perfusing angioplasty-damaged coronary arteries. Flow cytometric detection. Arterioscler Thromb. 1992 Dec;12(12):1475–1487. doi: 10.1161/01.atv.12.12.1475. [DOI] [PubMed] [Google Scholar]
  43. Shattil S. J., Cunningham M., Hoxie J. A. Detection of activated platelets in whole blood using activation-dependent monoclonal antibodies and flow cytometry. Blood. 1987 Jul;70(1):307–315. [PubMed] [Google Scholar]
  44. Stuard S., Carreno M. P., Poignet J. L., Albertazzi A., Haeffner-Cavaillon N. A major role for CD62P/CD15s interaction in leukocyte margination during hemodialysis. Kidney Int. 1995 Jul;48(1):93–102. doi: 10.1038/ki.1995.272. [DOI] [PubMed] [Google Scholar]
  45. Suzuki H., Miura S., Imaeda H., Suzuki M., Han J. Y., Mori M., Fukumura D., Tsuchiya M., Ishii H. Enhanced levels of chemiluminescence and platelet activating factor in urease-positive gastric ulcers. Free Radic Biol Med. 1996;20(3):449–454. doi: 10.1016/0891-5849(96)02048-5. [DOI] [PubMed] [Google Scholar]
  46. Tedder T. F., Steeber D. A., Chen A., Engel P. The selectins: vascular adhesion molecules. FASEB J. 1995 Jul;9(10):866–873. [PubMed] [Google Scholar]
  47. Tedder T. F., Steeber D. A., Pizcueta P. L-selectin-deficient mice have impaired leukocyte recruitment into inflammatory sites. J Exp Med. 1995 Jun 1;181(6):2259–2264. doi: 10.1084/jem.181.6.2259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Waddell T. K., Fialkow L., Chan C. K., Kishimoto T. K., Downey G. P. Signaling functions of L-selectin. Enhancement of tyrosine phosphorylation and activation of MAP kinase. J Biol Chem. 1995 Jun 23;270(25):15403–15411. doi: 10.1074/jbc.270.25.15403. [DOI] [PubMed] [Google Scholar]
  49. Wall J. E., Buijs-Wilts M., Arnold J. T., Wang W., White M. M., Jennings L. K., Jackson C. W. A flow cytometric assay using mepacrine for study of uptake and release of platelet dense granule contents. Br J Haematol. 1995 Feb;89(2):380–385. doi: 10.1111/j.1365-2141.1995.tb03315.x. [DOI] [PubMed] [Google Scholar]
  50. Wedzicha J. A., Syndercombe-Court D., Tan K. C. Increased platelet aggregate formation in patients with chronic airflow obstruction and hypoxaemia. Thorax. 1991 Jul;46(7):504–507. doi: 10.1136/thx.46.7.504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Whincup P. H., Mendall M. A., Perry I. J., Strachan D. P., Walker M. Prospective relations between Helicobacter pylori infection, coronary heart disease, and stroke in middle aged men. Heart. 1996 Jun;75(6):568–572. doi: 10.1136/hrt.75.6.568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wu K. K., Hoak J. C. A new method for the quantitative detection of platelet aggregates in patients with arterial insufficiency. Lancet. 1974 Oct 19;2(7886):924–926. doi: 10.1016/s0140-6736(74)91131-3. [DOI] [PubMed] [Google Scholar]
  53. Xiang Z., Censini S., Bayeli P. F., Telford J. L., Figura N., Rappuoli R., Covacci A. Analysis of expression of CagA and VacA virulence factors in 43 strains of Helicobacter pylori reveals that clinical isolates can be divided into two major types and that CagA is not necessary for expression of the vacuolating cytotoxin. Infect Immun. 1995 Jan;63(1):94–98. doi: 10.1128/iai.63.1.94-98.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Yoshida N., Granger D. N., Evans D. J., Jr, Evans D. G., Graham D. Y., Anderson D. C., Wolf R. E., Kvietys P. R. Mechanisms involved in Helicobacter pylori-induced inflammation. Gastroenterology. 1993 Nov;105(5):1431–1440. doi: 10.1016/0016-5085(93)90148-6. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES