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Decreased apoptosis of β 2 ‐ integrin‐deficient bovine neutrophils
Author(s) -
Nagahata Hajime,
Higuchi Hidetoshi,
Teraoka Hiroki,
Takahashi K,
Inanami Osamu,
Kuwabara Mikwori
Publication year - 2004
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1111/j.1440-1711.2004.01202.x
Subject(s) - cd18 , annexin , apoptosis , zymosan , stimulation , neutrophile , incubation , biology , microbiology and biotechnology , medicine , endocrinology , chemistry , integrin alpha m , biochemistry , in vitro , flow cytometry
Stimulant‐induced viability of neutrophils, nuclear‐fragmentation, increase in intracellular calcium ([Ca 2+ ] i ), expression of annexin V on neutrophils and proteolysis of a fluorogenic peptide substrate Ac‐DEVD‐MCA (acetyl Asp‐Glu‐Val‐Asp α‐[4‐methyl‐coumaryl‐7‐amide]) by neutrophil lysates from five normal calves and three calves with leucocyte adhesion deficiency were determined to evaluate the apoptosis of normal and CD18‐deficient neutrophils. Viability was markedly decreased in control neutrophils stimulated with opsonized zymosan (OPZ), compared to CD18‐deficient neutrophils at 37°C after incubation periods of 6 and 24 hours. The rate of apoptosis of control neutrophils stimulated with OPZ increased significantly depending on the incubation time, whereas no apparent increase in apoptosis was found in CD18‐deficient neutrophils under the same conditions. Aggregated bovine (Agg) IgG‐induced apoptosis of control neutrophils was not significantly different from that of CD18‐deficient neutrophils. The expression of annexin V on OPZ‐stimulated control neutrophils was greater than that of unstimulated ones 6 h after stimulation. No apparent increase in annexin V expression on CD18‐deficient neutrophils was found with OPZ stimulation. A delay in apoptosis was demonstrated in CD18‐deficient bovine neutrophils and this appeared to be closely associated with lowered signalling via [Ca 2+ ] i , diminished annexin V expression on the cell surface, and decreased caspase 3 activity in lysates.