A Differential Role for Cell Shape in Neutrophil Tethering and Rolling on Endothelial Selectins under Flow

Erik B. Finger, Richard E. Bruehl, Dorothy F. Bainton, Timothy A. Springer

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Abstract

We investigated the role of neutrophil microvilli in interactions with E-selectin and P-selectin in hydrodynamic shear flow by disruption with cytochalasin B, hypotonic swelling, and chilling. Cytochalasin B only marginally reduced microvilli numbers (from 30 ± 6 to 16 ± 6 per cell perimeter, p < 0.005) as shown by electron microscopy, completely disrupted tethering in shear flow to E-selectin and P-selectin, increased the strength of rolling adhesions on E-selectin and P-selectin, and increased cell deformability in shear flow with a likely increase in the area of cell:substrate contact. Hypoosmotic swelling markedly reduced microvilli number (to 6 ± 5 per perimeter, p < 0.005), almost completely inhibited tethering on E- and P-selectin, and increased the strength of rolling adhesions on P-selectin but not on E-selectin. Chilling almost completely abolished microvilli (to 3 ± 3 per perimeter, p < 0.005), but pseudopod-like structures were present, and had little effect on tethering in flow. Immunogold labeling of L-selectin, which is normally clustered on tips of microvilli, showed that in the absence of microvilli it remained in small clusters. Our studies show that alterations in cell morphology and viscoelasticity can have opposing effects on tethering and rolling, showing that they are independently regulatable. Furthermore, our results suggest that the association of molecules that mediate rolling with microvilli tips may be important not just to enhance presentation, but for other functions such as to promote resistance to extraction from the membrane or cooperative interactions among clustered receptors.

Original languageEnglish (US)
Pages (from-to)5085-5096
Number of pages12
JournalJournal of Immunology
Volume157
Issue number11
StatePublished - Dec 1 1996

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Selectins
Cell Shape
Microvilli
P-Selectin
E-Selectin
Neutrophils
Cytochalasin B
L-Selectin
Pseudopodia
Hydrodynamics
Electron Microscopy
Membranes

Cite this

A Differential Role for Cell Shape in Neutrophil Tethering and Rolling on Endothelial Selectins under Flow. / Finger, Erik B.; Bruehl, Richard E.; Bainton, Dorothy F.; Springer, Timothy A.

In: Journal of Immunology, Vol. 157, No. 11, 01.12.1996, p. 5085-5096.

Research output: Contribution to journalArticle

Finger, Erik B. ; Bruehl, Richard E. ; Bainton, Dorothy F. ; Springer, Timothy A. / A Differential Role for Cell Shape in Neutrophil Tethering and Rolling on Endothelial Selectins under Flow. In: Journal of Immunology. 1996 ; Vol. 157, No. 11. pp. 5085-5096.
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