Journal of Leukocyte Biology eBioscience full spectrum cell analysis
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Originally published online as doi:10.1189/jlb.0204077 on July 7, 2004

Published online before print July 7, 2004
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(Journal of Leukocyte Biology. 2004;76:594-602.)
© 2004 by Society for Leukocyte Biology

Direct quantification of the modulation of interaction between cell- or surface-bound LFA-1 and ICAM-1

Joana Vitte, Anne Pierres, Anne-Marie Benoliel and Pierre Bongrand1

Laboratoire d’Immunologie, INSERM U600, CNRS FRE 2059, Univ. Mediterranée, Hôpital de Ste-Marguerite, Marseille, Cedex, France

1 Correspondence: Laboratoire d'Immunologie, INSERM U600, CNRS FRE 2059, Univ. Mediterranée, Hôpital de Ste-Marguerite, 270 Bd de Ste-Marguerite, 13009 France. E-mail: bongrand{at}marseille.inserm.fr

The functional activity of leukocyte integrins is highly regulated by several mechanisms related to intrinsic molecular properties and receptor interaction with the cell membrane. Here, we present a microkinetic study of the lymphocyte function-associated antigen-1-mediated interaction between flowing Jurkat cells and surface- or cell-bound intercellular adhesion molecule-1 (ICAM-1). We conclude that adhesion is initiated by the formation of a single bond with ~0.3 s–1 dissociation rate, and attachment is subsequently strengthened by the formation of additional bonds during the next 10 s; exposing cells to Mg2+ or Mn2+ resulted in up to a 16-fold increase of the binding frequency, in line with reported measurements performed on isolated molecules with surface plasmon resonance methodology; cell-bound ICAM-1 molecules were more efficient in mediating adhesion than Fc-ICAM-1, properly oriented and bound by surface-adsorbed protein A; and quantitative analysis of binding frequency suggested that adhesion efficiency was ten- to 100-fold lower than the maximum value allowed by previously determined association rates of soluble molecules. It is concluded that the presented methodology provides a simple and unique way of dissecting the initial step of cell adhesion and discriminating between affinity and avidity modulation of adhesion receptors.

Key Words: laminar flow chamber • confocal microscopy • receptor topography • avidity • dissociation rate






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Copyright © 2004 by the Society for Leukocyte Biology.