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Analysis of the functional characteristics of L‐selectin and its expression on normal and CD18‐deficient bovine neutrophils
Author(s) -
Nagahata H,
Higuchi H,
Yamashiki N,
Yamaguchi M
Publication year - 2000
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 - 10.1046/j.1440-1711.2000.00916.x
Subject(s) - medicine , endocrinology , l selectin , cd18 , phorbol , superoxide , stimulation , n formylmethionine leucyl phenylalanine , neutrophile , chemistry , biology , phosphorylation , protein kinase c , inflammation , biochemistry , receptor , integrin alpha m , enzyme
In vivo responsiveness to epinephrine, expression of L‐selectin on neutrophils, changes in intracellular calcium ([Ca 2+ ] i ), sulfatide‐induced superoxide production and tyrosine phosphorylation in neutrophils were evaluated to elucidate the role of L‐selectin‐associated functions of normal and CD18‐deficient bovine neutrophils. The number of neutrophils in peripheral blood was significantly increased ( P < 0.05) in four normal calves at 5–20 min after in vivo administration of epinephrine; however, no significant increase of neutrophils was found in three calves with bovine leucocyte adhesion deficiency (BLAD). Expression of L‐selectin on neutrophils from three calves with BLAD was 61–77% of that of normal calves. Pretreatment of neutrophils with phorbol myristate acetate caused a marked decrease in the expression of L‐selectin on neutrophils from both normal and BLAD calves. The sulfatide‐induced sustained phase of [Ca 2+ ] i concentration in neutrophils from calves with BLAD was significantly ( P < 0.05) decreased. Following stimulation with aggregated IgG, the transient phase of [Ca 2+ ] i in neutrophils from normal and BLAD calves was increased; however, the sustained phase of [Ca 2+ ] i in BLAD neutrophils was significantly lower ( P < 0.05) than that of controls. Sulfatide‐induced O 2 − production and chemiluminescent response in neutrophils from calves with BLAD were 48–51% of those of normal calves and were inhibited by genistein and wortmannin, respectively, in a dose‐dependent manner. The amount of tyrosine phosphorylated 100 kDa protein in neutrophils from BLAD calves stimulated with sulfatides was 57% of that of controls. The degree of L‐selectin expression on neutrophils was correlated with the intracellular signalling events and the related superoxide production.