Analysis of the functional characteristics of L‐selectin and its expression on normal and CD18‐deficient bovine neutrophils

In vivo responsiveness to epinephrine, expression of L‐selectin on neutrophils, changes in intracellular calcium ([Ca2+]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 [Ca2+]i concentration in neutrophils from calves with BLAD was significantly (P < 0.05) decreased. Following stimulation with aggregated IgG, the transient phase of [Ca2+]i in neutrophils from normal and BLAD calves was increased; however, the sustained phase of [Ca2+]i in BLAD neutrophils was significantly lower (P < 0.05) than that of controls. Sulfatide‐induced O2− 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.

[1]  H. Nagahata,et al.  Neurtrophil function and pathologic findings in Holstein calves with leukocyte adhesion deficiency. , 1994, American journal of veterinary research.

[2]  G. Murphy,et al.  Metalloproteinase-mediated Regulation of L-selectin Levels on Leucocytes (*) , 1996, The Journal of Biological Chemistry.

[3]  C. Laudanna,et al.  Effect of Inhibitors of Distinct Signaling Pathways on Neutrophil O2− Generation in Response to Tumor Necrosis Factor-α, and Antibodies Against CD18 and CD11a: Evidence for a Common and Unique Pattern of Sensitivity to Wortmannin and Protein Tyrosine , 1993 .

[4]  Scott M. Seo,et al.  Signaling functions of L-selectin in neutrophils: alterations in the cytoskeleton and colocalization with CD18. , 1999, Journal of immunology.

[5]  T. Carlos,et al.  Leukocyte-endothelial adhesion molecules. , 1994, Blood.

[6]  A Etzioni,et al.  In vivo neutrophil and lymphocyte function studies in a patient with leukocyte adhesion deficiency type II. , 1994, Blood.

[7]  R. Larson,et al.  Structure and Function of Leukocyte Integrins , 1990, Immunological reviews.

[8]  J. Griffin,et al.  Regulation of leukocyte migration by activation of the leukocyte adhesion molecule-1 (LAM-1) selectin , 1991, Nature.

[9]  J D Chambers,et al.  Two-step model of leukocyte-endothelial cell interaction in inflammation: distinct roles for LECAM-1 and the leukocyte beta 2 integrins in vivo. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[10]  T. Springer,et al.  Leukocytes roll on a selectin at physiologic flow rates: Distinction from and prerequisite for adhesion through integrins , 1991, Cell.

[11]  J I Gallin,et al.  Current concepts: immunology. Neutrophils in human diseases. , 1987, The New England journal of medicine.

[12]  T. Springer,et al.  Leukocyte adhesion deficiency: an inherited defect in the Mac-1, LFA-1, and p150,95 glycoproteins. , 1987, Annual review of medicine.

[13]  R. Dixon,et al.  Tyrosine phosphatase antagonist-induced activation of the neutrophil NADPH oxidase: a possible role for protein kinase C. , 1995, Immunology.

[14]  N. Hogg.  The leukocyte integrins. , 1989, Immunology today.

[15]  R. Horst,et al.  Regulation of L‐selectin and CD18 on bovine neutrophils by glucocorticoids: effects of cortisol and dexamethasone , 1995, Journal of leukocyte biology.

[16]  Timothy A. Springer,et al.  Adhesion receptors of the immune system , 1990, Nature.

[17]  H. Nochi,et al.  Expression and role of adhesion molecule CD18 on bovine neutrophils. , 1995, Canadian journal of veterinary research = Revue canadienne de recherche veterinaire.

[18]  C. Laudanna,et al.  Effect of inhibitors of distinct signalling pathways on neutrophil Q2- generation in response to tumor necrosis factor-alpha, and antibodies against CD18 and CD11a: evidence for a common and unique pattern of sensitivity to wortmannin and protein tyrosine kinase inhibitors. , 1993, Biochemical and biophysical research communications.

[19]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[20]  H. Nochi,et al.  Biosynthesis of B2-integrin, intracellular calcium signalling and functional responses of normal and CD18-deficient bovine neutrophils. , 1996, Research in veterinary science.

[21]  S. Simon,et al.  Beta 2-integrin and L-selectin are obligatory receptors in neutrophil aggregation [see comments] , 1993 .

[22]  K. Ley,et al.  Lectin-like cell adhesion molecule 1 mediates leukocyte rolling in mesenteric venules in vivo. , 1991, Blood.