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Double Tether Extraction from Human Neutrophils and Its Comparison with CD4⁺ T-Lymphocytes

Department of Biomedical Engineering, Washington University, St. Louis, Missouri

Correspondence: Address reprint requests to Jin-Yu Shao, PhD, Dept. of Biomedical Engineering, Washington University in St. Louis, Campus Box 1097, Rm. 290E, Uncas A. Whitaker Hall, 1 Brookings Dr., St. Louis, MO 63130-4899. Tel.: 314-935-7467; Fax: 314-935-7448; E-mail: shao{at}biomed.wustl.edu.

The initial arrest and subsequent rolling of a leukocyte on the vascular endothelium is believed to be facilitated by the extraction of tethers, which are narrow membranous tubes drawn from the leukocyte. Although single tether extraction from neutrophils has been studied thoroughly, the relationship between the tether force (F) and tether-growth velocity (U_t) is still unknown for double tethers drawn from neutrophils. In this study, we have determined this relationship with the micropipette-aspiration technique. As a comparison, tether extraction from CD4⁺ T-lymphocytes was also studied. The threshold force and effective viscosity for single tether extraction from passive CD4⁺ T-lymphocytes were found to be 46 pN and 1.55 pN · s/µm, respectively. These values were modulated by stimulation with phorbol myristate acetate (PMA), but not interleukin-8 (IL-8). More importantly, for both types of leukocyte, the threshold force and effective viscosity for double tether extraction are about twice as large as those corresponding to single tether extraction. Neither IL-8 nor PMA stimulation had any effect on this correlation. These results indicate that double tethers are highly localized on cellular surfaces and independent of each other during the rolling process.

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