Date of Award

1986

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Molecular and Cellular Biology and Pathobiology

College

College of Graduate Studies

First Advisor

Harvey L. Bank

Second Advisor

Armand Glassman

Third Advisor

Harold Hempling

Fourth Advisor

Gary Landreth

Fifth Advisor

Jo Anne Simpson

Sixth Advisor

Michael Weise

Abstract

When human blood neutrophils are isolated for the circulation, they begin to lose chemotactic responsiveness. To characterize this loss of function which occurs during storage, an enhanced videomicroscope system was developed and computer-assisted image analysis was utilized to perform kinematic analysis on neutrophils migrating in an agarose assay system. The locomotor behavior of neutrophils was measured in response to a gradient of chemoattractant (chemotaxis) and in response to an isotropic distribution of chemoattractant (chemokinesis). During chemotaxis, neutrophils migrate at ~20 um/min, make few turns and orient toward a chemoattractant with high accuracy and precision. During chemokinesis, neutrophils migrate at a slower speed, change direction more often and show no directional preference. The vast majority of normal, fresh neutrophils show chemotaxis in response to a gradient, while ~10% of fresh normal cell fail to orient in a gradient. After storage, ~65% of the cells show normal chemotaxis in response to a gradient. The other 35% migrate at 8-9um/min and show no directional preference; they orient randomly. This behavior is quantitatively similar to chemokinesis and suggests that such cells either cannot detect the gradient or cannot respond to the gradient. A model is proposed which suggests that the ability to orient accurately in a gradient is a function of a structural asymetry within the cytoskeleton and that the loss of this unique structural arrangement by some cells during storage produces slower I non-oriented “chemokinetic” locomotion in response to a gradient.

Rights

All rights reserved. Copyright is held by the author.

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