TY - JOUR
T1 - Rolling on E- or P-selectin induces the extended but not high-affinity conformation of LFA-1 in neutrophils
AU - Kuwano, Yoshihiro
AU - Spelten, Oliver
AU - Zhang, Hong
AU - Ley, Klaus
AU - Zarbock, Alexander
PY - 2010/7/29
Y1 - 2010/7/29
N2 - Human blood neutrophils rolling on E- or P-selectin reduced their rolling velocity when intercellular adhesion molecule (ICAM)-1 was available. Similar to mouse neutrophils, this was dependent on P-selectin glycoprotein ligand 1 (PSGL1), αLβ2 integrin, the Src family tyrosine kinase FGR and spleen tyrosine kinase SYK. Blocking phospholipase C or p38 MAP kinase attenuated, but did not abolish the velocity reduction. To test expression of integrin activation epitopes, we adapted an immobilized reporter assay and developed a new homogeneous microfluidics-based reporter antibody binding assay. Rolling on E- or P-selectin induced the extension reporter epitopes KIM127 and NKI-L16, but not the high affinity reporter epitope monoclonal antibody (mAb) 24. This enabled rolling neutrophils to bind to immobilized extension reporter, but not activation reporter antibodies and allowed binding of soluble KIM127 during rolling. We conclude that human neutrophil rolling on E- or P-selectin induces the extended α Lβ2 integrin conformation through signaling triggered by PSGL-1 engagement.
AB - Human blood neutrophils rolling on E- or P-selectin reduced their rolling velocity when intercellular adhesion molecule (ICAM)-1 was available. Similar to mouse neutrophils, this was dependent on P-selectin glycoprotein ligand 1 (PSGL1), αLβ2 integrin, the Src family tyrosine kinase FGR and spleen tyrosine kinase SYK. Blocking phospholipase C or p38 MAP kinase attenuated, but did not abolish the velocity reduction. To test expression of integrin activation epitopes, we adapted an immobilized reporter assay and developed a new homogeneous microfluidics-based reporter antibody binding assay. Rolling on E- or P-selectin induced the extension reporter epitopes KIM127 and NKI-L16, but not the high affinity reporter epitope monoclonal antibody (mAb) 24. This enabled rolling neutrophils to bind to immobilized extension reporter, but not activation reporter antibodies and allowed binding of soluble KIM127 during rolling. We conclude that human neutrophil rolling on E- or P-selectin induces the extended α Lβ2 integrin conformation through signaling triggered by PSGL-1 engagement.
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U2 - 10.1182/blood-2010-01-266122
DO - 10.1182/blood-2010-01-266122
M3 - Article
C2 - 20445017
AN - SCOPUS:77955454503
SN - 0006-4971
VL - 116
SP - 617
EP - 624
JO - Blood
JF - Blood
IS - 4
ER -