Abstract
Upon injury, stable thrombi formation requires the recruitment of platelets, leukocytes, and various clotting factors, to provide sufficient inhibition of hemostasis. Classical models of thrombosis involve either ex vivo isolation of platelets and subsequent quantification of aggregation through light transmission aggregometry or in vivo murine intravital thrombosis models (laser injury, ferric chloride, or rose Bengal). Flow adhesion models allow for accurate quantification of the contribution of cell-types to thrombi formation. Here, we describe the use of flow chambers to flow human blood over activated endothelial cells to observe leukocyte-endothelial adhesion at arterial and venous shear rates.
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References
Pirmohamed M (2006) Warfarin: almost 60 years old and still causing problems. Br J Clin Pharmacol 62(5):509–511. https://doi.org/10.1111/j.1365-2125.2006.02806.x
Swystun LL, Liaw PC (2016) The role of leukocytes in thrombosis. Blood 128(6):753–762. https://doi.org/10.1182/blood-2016-05-718114
Rayes J, Bourne JH, Brill A, Watson SP (2020) The dual role of platelet-innate immune cell interactions in thrombo-inflammation. Res Pract Thromb Haemost 4(1):23–35. https://doi.org/10.1002/rth2.12266
Munro JM, Lo SK, Corless C, Robertson MJ, Lee NC, Barnhill RL et al (1992) Expression of sialyl-Lewis X, an E-selectin ligand, in inflammation, immune processes, and lymphoid tissues. Am J Pathol 141(6):1397–1408
Huo Y, Xia L (2009) P-selectin glycoprotein ligand-1 plays a crucial role in the selective recruitment of leukocytes into the atherosclerotic arterial wall. Trends Cardiovasc Med 19(4):140–145. https://doi.org/10.1016/j.tcm.2009.07.006
Woodman RC, Johnston B, Hickey MJ, Teoh D, Reinhardt P, Poon BY et al (1998) The functional paradox of CD43 in leukocyte recruitment: a study using CD43-deficient mice. J Exp Med 188(11):2181–2186. https://doi.org/10.1084/jem.188.11.2181
McDonald B, Kubes P (2015) Interactions between CD44 and hyaluronan in leukocyte trafficking. Front Immunol 6:68. https://doi.org/10.3389/fimmu.2015.00068
Shamri R, Grabovsky V, Gauguet JM, Feigelson S, Manevich E, Kolanus W et al (2005) Lymphocyte arrest requires instantaneous induction of an extended LFA-1 conformation mediated by endothelium-bound chemokines. Nat Immunol 6(5):497–506. https://doi.org/10.1038/ni1194
Lawson C, Ainsworth M, Yacoub M, Rose M (1999) Ligation of ICAM-1 on endothelial cells leads to expression of VCAM-1 via a nuclear factor-kappaB-independent mechanism. J Immunol 162(5):2990–2996
Kattula S, Byrnes JR, Wolberg AS (2017) Fibrinogen and Fibrin in Hemostasis and Thrombosis. Arterioscler Thromb Vasc Biol 37(3):e13–e21. https://doi.org/10.1161/ATVBAHA.117.308564
Acknowledgments
This work was supported by a college-funded studentship from the University of Birmingham and the Centre of Membrane Proteins and Receptors.
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Bourne, J.H. (2022). Quantification of Leukocyte–Endothelial Cell Interaction During In Vitro Models of Thrombosis at Arterial and Venous Shear Rates. In: Benest, A.V. (eds) Angiogenesis. Methods in Molecular Biology, vol 2441. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2059-5_22
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DOI: https://doi.org/10.1007/978-1-0716-2059-5_22
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