Decreased apoptosis of beta 2- integrin-deficient bovine neutrophils.
Stimulant-induced viability of neutrophils, nuclear-fragmentation, increase in intracellular calcium ([Ca2+]i), expression of annexin V on neutrophils and proteolysis of a fluorogenic peptide substrate Ac-DEVD-MCA (acetyl Asp-Glu-Val-Asp alpha-[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 degrees 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 [Ca2+]i, diminished annexin V expression on the cell surface, and decreased caspase 3 activity in lysates.
Department of Animal Health, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501, Japan. nght-h@rakuno.ac.jp
Hajime
H
Nagahata
Hidetoshi
H
Higuchi
Hiroki
H
Teraoka
Kenji
K
Takahashi
Kensi
K
Takahashi
Mikinori
M
Kuwabara
Osamu
O
Inanami
Mikwori
M
Kuwabara
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D018821:CD18 Antigens / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002470:Cell Survival; D005456:Fluorescent Dyes / Q000378:metabolism; D007074:Immunoglobulin G / Q000494:pharmacology; D018370:Leukocyte-Adhesion Deficiency Syndrome / Q000276:immunology / Q000378:metabolism* / Q000473:pathology*; D018375:Neutrophil Activation; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D010455:Peptides / Q000378:metabolism; D015054:Zymosan / Q000494:pharmacology
D017304:Annexin A5; D018821:CD18 Antigens; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D010455:Peptides; D015054:Zymosan; D002118:Calcium