TY - JOUR
T1 - Analysis of polarization and orientation of human polymorphonuclear leukocytes by computer-interfaced video microscopy
AU - MacFarlane, G. D.
AU - Herzberg, M. C.
AU - Nelson, R. D.
N1 - Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 1987
Y1 - 1987
N2 - Chemotactic behavior is a complex culture response to chemical environmental stimuli. For polymorphonuclear leukocytes (PMNs), such behavior involves net migration as well as changes in cell shape and cell orientation. Accordingly, we have applied computer-interfaced video microscopy to analyze cell shape and orientation in control and patient PMNs migrating under agarose. From a digitized tracing of the PMNs at the leading front of migration, cells were characterized in terms of area, circumference, and longest dimension. A shape factor and angle of orientation were computed. Numerical shape factors discriminated three PMN morphologies: polar, apolar, and hyperpolar. Only polar cells could be oriented. Orientation of polar cells was defined as toward, away, or disoriented with respect to the chemotactic gradient. Apolar cells were considered to be nonoriented. Of PMNs from healthy controls, 30 ± 5% of the cells were oriented toward and 11 ± 4% of the cells were oriented away from the gradient. For PMNs from patients with localized juvenile periodontitis, a 40% deficit in net migration was associated with reduced orientation toward (5 ± 2%) and elevated orientation away from the gradient (33 ± 9%). PMNs from a panel of patients with thermal injury showed reduced migration and orientation the gradient associated with elevated percentages of apolar cells. Such analysis of PMN polarization and orientation of the leading front permitted calculation of a chemotactic behavior index. Application of this multiparameter index to the analysis of the chemotactic response may identify PMNs that are defective, but not by evaluation of any single variable.
AB - Chemotactic behavior is a complex culture response to chemical environmental stimuli. For polymorphonuclear leukocytes (PMNs), such behavior involves net migration as well as changes in cell shape and cell orientation. Accordingly, we have applied computer-interfaced video microscopy to analyze cell shape and orientation in control and patient PMNs migrating under agarose. From a digitized tracing of the PMNs at the leading front of migration, cells were characterized in terms of area, circumference, and longest dimension. A shape factor and angle of orientation were computed. Numerical shape factors discriminated three PMN morphologies: polar, apolar, and hyperpolar. Only polar cells could be oriented. Orientation of polar cells was defined as toward, away, or disoriented with respect to the chemotactic gradient. Apolar cells were considered to be nonoriented. Of PMNs from healthy controls, 30 ± 5% of the cells were oriented toward and 11 ± 4% of the cells were oriented away from the gradient. For PMNs from patients with localized juvenile periodontitis, a 40% deficit in net migration was associated with reduced orientation toward (5 ± 2%) and elevated orientation away from the gradient (33 ± 9%). PMNs from a panel of patients with thermal injury showed reduced migration and orientation the gradient associated with elevated percentages of apolar cells. Such analysis of PMN polarization and orientation of the leading front permitted calculation of a chemotactic behavior index. Application of this multiparameter index to the analysis of the chemotactic response may identify PMNs that are defective, but not by evaluation of any single variable.
UR - http://www.scopus.com/inward/record.url?scp=0023212446&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0023212446&partnerID=8YFLogxK
U2 - 10.1002/jlb.41.4.307
DO - 10.1002/jlb.41.4.307
M3 - Article
C2 - 3471828
AN - SCOPUS:0023212446
SN - 0741-5400
VL - 41
SP - 307
EP - 317
JO - Journal of Leukocyte Biology
JF - Journal of Leukocyte Biology
IS - 4
ER -