Rapid flow cytometry method for quantitation of LFA-1-adhesive T cells

Clinical and Vaccine Immunology : CVI
B E CrucianC F Sams

Abstract

Adhesion molecules are important for leukocyte endothelial attachment and migration to sites of inflammation. The LFA-1 (CD11a and CD18) integrin molecule is constitutively expressed on the T-cell surface. Following T-cell activation, a rapid conformational change of LFA-1 to an "adhesive" state occurs, allowing LFA-1 binding to intracellular cell adhesion molecule type 1 (ICAM-1)-expressing targets, such as antigen-presenting cells. For this study, a rapid flow cytometry method for the quantitation of LFA-1-adhesive T cells following activation was developed. Purified ICAM-1 was bound to 4.5-microm-diameter beads. Following peripheral blood mononuclear cell activation culture (phorbol myristate acetate and ionomycin), the cells were incubated with the ICAM-1 beads, which allowed attachment to occur. The T cell-bead complexes were then resolved from unbound T cells by flow cytometry. Multicolor analysis allowed a complete phenotypic analysis of the adhesive T-cell subsets. Experimental controls indicated that the T cell-bead attachment was LFA-1 and ICAM-1 specific. Very little binding between unactivated T cells and ICAM beads or between activated T cells and plain beads was observed. The kinetics of the response was extremely...Continue Reading

Citations

Nov 21, 2007·Biological Chemistry·Karen Hänel, Dieter Willbold
Feb 25, 2009·Immunopharmacology and Immunotoxicology·Chen WeihuaGao Chunji
Nov 3, 2010·The Journal of Immunology : Official Journal of the American Association of Immunologists·Ronen SumaginIngrid H Sarelius
Dec 12, 2019·Frontiers in Chemistry·Md Amran HowladerChristopher W Cairo

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