CHARACTERIZATION OF MORPHOLOGICAL CHANGES IN ACTIVATED NEUTROPHILS
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Date
2005-05
Department
Hood College Biology
Program
Biomedical and Environmental Science
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Abstract
Neutrophils leave the vasculature and migrate along a chemotactic gradient to an
infectious focus where they ingest invading microorganisms. To accomplish this myriad
of functions, neutrophils undergo considerable morphological changes. Treatment of
neutrophils with the chemotactic peptide, formyl-methionyl-leucyl-phenylalanine
(fMLF), 5 x 10(-9) M, results in a change in cellular morphology that includes formation of
agranular projections and polarization of the cell. As determined by flow cytometric
analysis these morphological change occurs within 5 min, peaks by 30 min, and are
accompanied by a change in forward and right angle light scatter. Fibrinogen prolongs
the fMLF-induced response through 4 h. Changes in right angle light scatter parallel
morphological change observed by light microscopy, indicating that flow cytometry can
also be used to evaluate changes in cell morphology. Other chemoattractants were less
effective at inducing a change in morphology — fMLF >> LTB4 = C5a > 1L-8 > PAF =
LPS. Disruption of microtubules with colchicine altered forward light scatter and cell
polarization while the disruption of microfilaments with cyto B abrogates all
morphological alterations, demonstrating the involvement of the cytoskeleton in the
chemotactic process of neutrophils.