Purification and characterisation of a steroid-induced modulator of neutrophil function

Young, John (1998) Purification and characterisation of a steroid-induced modulator of neutrophil function. PhD thesis, University of Glasgow.

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Printed Thesis Information: https://eleanor.lib.gla.ac.uk/record=b1713501

Abstract

A monocyte-derived factor that stimulates the locomotion of human neutrophils on an albumin-coated glass surface has been prepared from the culture supernatant of dexamethasone-treated human monocytes and called STMS (steroid-treated monocyte supernatant). A modified cell tracking program has been developed and the parameters of locomotion determined by the analysis of Gail and Boone (1970, 1972) for cells moving in a persistent random walk. Cells moving in uniform concentrations of STMS, interleukin-8 (IL-8) and n-formyl-methionyl- leucyl-phenylalanine (fMLP) chosen to give a sub-maximal speed of locomotion show persistent, random and constrained random diffusion respectively with augmented diffusion coefficients of 0.8 ± 0.1, 0.14 ± 0.02 and 0.12 ± 0.03 μm2 per second for STMS, IL-8 and fMLP respectively. The augmented diffusion coefficient and the underlying persistence are therefore sensitive quantitative assay parameters for STMS activity and the qualitative characteristics of locomotion allow STMS activity to be distinguished from that of all other factors tested. The contribution of lowered adhesion to locomotion was examined in a novel tilt- assay which demonstrated that cells in the presence of STMS, but not other factors, moved down slope with significantly increased speed while maintaining contact with the substratum. The results were interpreted in terms of the bipolar form of STMS-treated cells, contrasting with multipolar forms in response to other agents. This together with low adhesiveness plus an inherent tendency of a single locomotor focus to continue motion in its original direction has been used to explain the difference between response to STMS and other factors. STMS has been proposed to prevent directed locomotion of neutrophils to an inflammatory site or to promote dispersive locomotion away from such a site, and perhaps to inhibit neutrophil transmigration between endothelial cells. STMS also inhibits adhesion of neutrophils to bovine aorta and human endothelial cells by a yet undetermined mechanism. It induces unique changes in neutrophil shape with a characteristic monopole of F-actin distribution, which may correlate with the dispersive locomotion observed in the absence of a concentration gradient. This factor also inhibits n-formyl-methionyl-leucyl-phenylalanine-induced chemotaxis of neutrophils in a modified Boyden chamber assay. The reduction of adhesion and the inhibition of chemotaxis by the factor in vitro indicate a possible in vivo anti-inflammatory role.

Item Type: Thesis (PhD)
Colleges/Schools: College of Medical Veterinary and Life Sciences
Supervisor's Name: Lawrance, Dr. A. J.
Date of Award: 1998
Depositing User: Mrs Monika Milewska-Fiertek
Unique ID: glathesis:1998-38919
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 09 Nov 2018 12:49
Last Modified: 09 Nov 2018 12:49
URI: http://theses.gla.ac.uk/id/eprint/38919
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