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Functional Maturation during Bovine Granulopoiesis

Ghent University, Faculty of Veterinary Medicine Department of Physiology, Biochemistry and Biometrics Salisburylaan 133 9820 Merelbeke, Belgium

Corresponding author:
C. Burvenich; e-mail:
christian.burvenich{at}rug.ac.be.

Granulocytic precursor cells undergo morphologic changes in the nucleus and the cytoplasm during the process of granulopoiesis, which takes place in the bone marrow. These changes are associated with the development of stage-specific proteins necessary for the highly specialized roles of polymorphonuclear leukocytes in phagocytosis, bacterial killing, and in mediating the inflammatory process. The objective of the current study was to sequence the various events that occur upon functional development of granulocytic bone marrow cells in the bovine species.

Cells were obtained from the bone marrow of clinically healthy cows and separated into different stages of maturation using density gradient centrifugation. Three cellular fractions were obtained that were enriched for either early immature, late immature or mature granulocytic cells. Functions and receptor expressions assessed in the three maturation stages were: Fc-IgG₂ receptor and CD11b expression, phagocytosis of Escherichia coli, respiratory burst activity, and cellular myeloperoxidase activity. Immature cells expressed already Fc-IgG₂ receptor and CD11b on their cytoplasma membrane. Phagocytic ability was acquired in the myelocytic stage, but only the more mature forms were readily capable of phagocytosis. Promyelocytes, myelocytes and metamyelocytes showed no respiratory burst activity. Only band and segmented cells produced reactive oxygen species. Myeloperoxidase was present at all stages of maturity. Thus, each of the maturation stages was characterized by a selective expression of one or more functions and receptors. Therefore, sequential biochemical maturation is postulated during bovine granulopoiesis.

Key Words: bovine • bone marrow • functional maturation • granulopoiesis

Abbreviation key: FBS = fetal bovine serum, FITC = fluorescein isothiocyanate, FS/SS = forward scatter side scatter, F = fluorescence, MFI = mean fluorescence intensity, MPO = myeloperoxidase, PI = propidium iodide, PMA = phorbol 12-myristate 13-acetate, PMN = polymorphonuclear leukocytes, ROS = reactive oxygen species

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