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*
Speros P. Martel Laboratory of Leukocyte Biology, Department of Pediatrics,
Department of Molecular and Human Genetics, and
Department of Medicine, Baylor College of Medicine, Houston, TX 77030;
§
Institute of Biosciences and Bioengineering, Rice University, Houston, TX;
¶
Department of Comparative Medicine, University of Alabama, Birmingham, AL 35294; and
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Department of Pathology, Washington University School of Medicine, St. Louis, MO 63110
To differentiate the unique and overlapping functions of LFA-1 and
Mac-1, LFA-1-deficient mice were developed by targeted homologous
recombination in embryonic stem cells, and neutrophil function was
compared in vitro and in vivo with Mac-1-deficient, CD18-deficient, and
wild-type mice. LFA-1-deficient mice exhibit leukocytosis but do not
develop spontaneous infections, in contrast to CD18-deficient mice.
After zymosan-activated serum stimulation, LFA-1-deficient neutrophils
demonstrated activation, evidenced by up-regulation of surface Mac-1,
but did not show increased adhesion to purified ICAM-1 or endothelial
cells, similar to CD18-deficient neutrophils. Adhesion of
Mac-1-deficient neutrophils significantly increased with stimulation,
although adhesion was lower than for wild-type neutrophils. Evaluation
of the strength of adhesion through LFA-1, Mac-1, and CD18 indicated a
marked reduction in firm attachment, with increasing shear stress in
LFA-1-deficient neutrophils, similar to CD18-deficient neutrophils, and
only a modest reduction in Mac-1-deficient neutrophils. Leukocyte
influx in a subcutaneous air pouch in response to TNF-
was reduced
by 67% and 59% in LFA-1- and CD18-deficient mice but increased by
198% in Mac-1-deficient mice. Genetic deficiencies demonstrate that
both LFA-1 and Mac-1 contribute to adhesion of neutrophils to
endothelial cells and ICAM-1, but adhesion through LFA-1 overshadows
the contribution from Mac-1. Neutrophil extravasation in response to
TNF- in LFA-1-deficient mice dramatically decreased, whereas
neutrophil extravasation in Mac-1-deficient mice markedly
increased.
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