Platelets enhance CD4+ lymphocyte adhesion to extracellular matrix under flow conditions: Role of platelet aggregation, integrins, and non-integrin receptors

Summary

The purpose of this study was to examine the role of platelets in CD4+ T lymphocyte adhesion to subendothelial extracellular matrix (ECM). Herpesvirus saimiri (HVS)-infected CD4+ T cells were incubated on ECM. An image analysis was used to evaluate T cell adhesion. Under static condition, T cell activation with 4-α-Phorbol 12-myristate 13-acetate (PMA) resulted in a 2.6-fold increase in cell adhesion. However, adhesion was not affected by platelets. In contrast, under flow (200s⁻¹), platelets markedly enhanced both resting and PMA-activatedT cell adhesion (33- and 48-fold), forming lymphocyte-platelet co-aggregates that contain approximately 90% of the adherent T cells. Abrogation of platelet aggregation with tirofiban inhibited formation of platelet-T cell co-aggregates under flow and reduced T cell adhesion by 74%. Separate and combined blockade of CD40L and P-selectin glycoprotein-1 (PSGL-1) on PMA-activated lymphocytes reduced adhesion under flow in the presence of platelets by 28%, 33%, and 55%, respectively. Blockade of β₁-integrins decreased adhesion under both static and flow conditions (by 35% and 44%, respectively), while blockade of β₂-integrin reduced adhesion only under static condition (by 23%). A similar adhesion pattern was observed using CD4+ T cells isolated from normal donor peripheral blood. In conclusion, platelets support CD4+ lymphocyte adhesion to ECM under flow by formation of heterotypic platelet-lymphocyte co-aggregates involving αII_bβ3 integrin and β₁-related integrins, as well as CD40L and PSGL-1.

Keywords

CD4+ T cells - platelets - integrins - CD40L - PSGL-1

• References

• 1 Weyrich AS, Lindemann S, Zimmerman CA. The evolving role of platelets in inflammation. J Thromb Haemost 2003; 01: 1897-905.
• 2 Theilmeier G, Remacle LenaertsT. et al. Circulating activated platelets assist TNP-1 monocytoid/endothelial cell interaction under shear stress. Blood 1999; 94: 2725-56.
• 3 Hagberg IA, Lyberg T. Evaluation of circulating platelet-leukocyte conjugates: a sensitive flow cytometric assay well suited for clinical studies. Platelets 2000; 11: 151-60.
• 4 Nishijima K, Kiryu J, Tsujikawa A. et al. Platelets adhering to the vascular wall mediate postischemic leukocyte-endothelial cell interactions in retinal microcirculation. Invest Ophthalmol Vis Sci 2004; 45: 977-84.
• 5 Rinder HM, Bonan JL, Rinder CS. et al. Dynamics of leukocyte-platelet adhesion in whole blood. Blood 1991; 78: 1730-7.
• 6 Li N, Hu H, Lindqvist M. et al. Platelet-leucocyte cross talk in whole blood. Arterioscler Thromb Vasc Biol 2000; 20: 2702-8.
• 7 da Costa PMartins, van den Berk N, Ulfman LH. et al. Platelet-monocyte complexes support monocyte adhesion to endothelium by enhancing secondary tethering and cluster formation. Arterioscler Thromb Vasc Biol 2004; 24: 193-9.
• 8 Hamburger SA, McEver RP. GMP-140 mediates adhesion of stimulated platelets to neutrophils. Blood 1990; 75: 550-4.
• 9 Konstantopoulos K, Neelamegham S, Burns AR. et al. Venous levels of shear support neutrophil-platelet adhesion and neutrophil aggregation in blood via P-selectin and β₂-integrin. Circulation 1998; 98: 873-82.
• 10 Diacovo TG, Roth SJ, Buccola JM. et al. Neutrophil rolling, arrest, and transmigration across activated, surface adherent platelets via sequential action of P-selectin and the β₂-integrin CD11b/CD18. Blood 1996; 88: 146-57.
• 11 Liu L, Kubes P. Molecular mechanisms of leukocyte recruitment: organ-specific mechanisms of action. Thromb Haemost 2003; 89: 213-20.
• 12 Henn V, Slupsky JR, Grafe M. et al. CD40 ligand on activated platelets triggers an inflammatory reaction of endothelial cells. Nature 1998; 391: 591-4.
• 13 Inwald DP, McDowall A, Peters MJ. et al. CD40 is constitutively expressed on platelets and provides a novel mechanism for platelet activation. Circ Res 2003; 92: 1041-8.
• 14 Vitkovsky YA, Kuznik BI, Solpov AV. Phenomenon of lymphocyte-platelet rosette forming. Immunology (Russian) 1999; 04: 35-7.
• 15 Diacovo TG, Puri KD, Warnock RA. et al. Platelet-mediated lymphocyte delivery to high endothelial venules. Science 1996; 273: 252-5.
• 16 Czyz A, Kolacz E, Angerer D. et al. Expression of activation antigens on lymphocyte surface and circulating platelet-leukocyte aggregates in ischemic heart disease. Kardiol Pol 2005; 62: 189-202.
• 17 Buchner K, Henn V, Grafe M. et al. CD40 ligand is selectively expressed on CD4+ T cells and platelets: implications for CD40-CD40L signaling in atherosclerosis. J Pathol 2003; 201: 288-95.
• 18 Danese S, Motte C, Reyes BMR. et al. Cutting edge: T cells trigger CD40-dependent platelet activation and granular RANTES release:A novel pathway for immune response amplification. J Immunol 2004; 172: 2011-5.
• 19 Lalor P, Nash GB. Adhesion of flowing leucocytes to immobilized platelets. Br J Haematol 1995; 89: 725-32.
• 20 Hu H, Varon D, Hjemdahl P. et al. Platelet-leukocyte aggregation under shear stress: differential involvement of selectins and integrins. Thromb Haemost 2003; 90: 679-87.
• 21 Saha K, Sova P, Chao W. et al. Generation of CD4+ and CD8+ T-cell clones from PBLs of HIV-1 infected subjects using herpesvirus saimiri. Nat Med 1996; 02: 1272-5.
• 22 Bank I, Koltakov A, Nir-Glickman E. et al. Lovastatin and phospholipase Cγ regulate constitutive and protein kinase C dependent integrin mediated interactions of human T-cells with collagen. Cell Immunol 2003; 223: 35-45.
• 23 Gospodarowicz D, Greenburg G, Foidart JM. et al. The production and localization of laminin in cultured vascular and corneal endothelial cells. J Cell Physiol 1981; 107: 171-83.
• 24 Varon D, Dardik R, Shenkman B. et al. A new method for quantitative analysis of whole blood platelet interaction with extracellular matrix under flow conditions. Thromb Res 1997; 85: 283-94.
• 25 Li N, Goodall AH, Hjemdahl P. Efficient flow cytometric assay for platelet-leukocyte aggregates in whole blood using fluorescence signal triggering. Cytometry 1999; 35: 154-61.
• 26 Nurden AT, Nurden P. GPIIb/IIIa antagonists and other anti-integrins. Semin Vasc Med 2003; 03: 123-30.
• 27 McEver RP. Selectins: novel receptors that mediate leukocyte adhesion during inflammation. Thromb Haemos 1991; 65: 223-8.
• 28 Zhao L, Bath PM, May J. et al. P-selectin, tissue factor and CD40 ligand expression on platelet-leucocyte conjugates in the presence of a GPIIb/IIIa antagonist. Platelets 2003; 14: 473-80.
• 29 Aukrust P, Muller F, Ueland T. et al. Enhanced levels of soluble and membrane-bound CD40 ligand in patients with unstable angina. Possible reflection ofT lymphocyte and platelet involvement in the pathogenesis of acute coronary syndromes. Circulation 1999; 100: 614-20.
• 30 Wang CL, Wu YT, Liu CA. et al. Expression of CD40 ligand on CD4+ T-cells and platelets correlated to the coronary artery lesion and disease progress in Kawasaki disease. Pediatrics 2003; 111: E140-E147.
• 31 Schmidtke DW, Diamond SL. Direct observation of membrane tethers formed during neutrophil attachment to platelets or P-selectin under physiological flow. J Cell Biol 2000; 149: 719-30.
• 32 Evangelista V, Manarini S, Rotondo S. et al. Platelet/polymorphonuclear leukocyte interaction in dynamic conditions: evidence of adhesion cascade and cross talk between P-selectin and the β₂ integrin CD11b/CD18. Blood 1996; 88: 4183-94.
• 33 Kowalska MA, Ratajczak MZ, Majka M. et al. Stromal cell-derived factor-1 and macrophage-derived chemokine: 2 chemokines that activate platelets. Blood 2000; 96: 50-7.
• 34 Abi-Younes S, Si-Taha M, Luster AD. The CC chemokines MDC and TARC induce platelet activation via CCR4. Thromb Res 2001; 101: 279-89.
• 35 Campbell JJ, Hedrick J, Zlotnik A. et al. Chemokines and the arrest of lymphocytes rolling under flow conditions. Science 1998; 279: 381-4.
• 36 Gear AR, Camerini D. Platelet chemokines and chemokine receptors: linking hemostasis, inflammation, and host defence. Microcirc 2003; 10: 335-50.