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Twelve nulliparous Holstein heifers, approximately 12 months of age, were given intramammary injections of 5 μg Escherichia coli endotoxin in sterile saline. Peripheral blood and mammary neutrophils were collected 14 h post-injection and evaluated for endogenous binding of immunoglobulins G1, G2, A and M. Isolated blood neutrophils from four of the above heifers were placed into a modified Boyden chemotaxis chamber, allowed to migrate for 3 h, and then evaluated for endogenous immunoglobulin binding. Phagocytosis was evaluated flow cytometrically for neutrophils from four heifers of the same group. Comparisons were made between neutrophils isolated from peripheral blood and neutrophils which had undergone chemotaxis in vitro. Percentage of neutrophils fluorescing and relative fluorescence intensity of endogenous binding of IgG1, and IgG2 were marginally increased (P<0.10) and IgM was significantly decreased (P<0.05) for mammary gland neutrophils relative to blood. The percentage of neutrophils binding IgG1, IgG2, IgA and IgM for blood and mammary neutrophils was 9.8%, 3.7%, 3.5% and 76.4% and 16.6%, 7.8%, 6.8% and 1.9%, respectively. Comparisons of neutrophils before and after chemotaxis in vitro revealed no significant changes in the percentage of neutrophils binding various immunoglobulins or relative fluorescence intensity, with the exception of IgM binding. Similar to results in vivo endogenous IgM binding was significantly depressed after chemotaxis. It was demonstrated that neutrophils which had migrated through the chemotaxis chamber had significantly higher percentages phagocytosing and adhering bacteria than did peripheral blood neutrophils. There were no differences in bacteria associated per neutrophil between cells from either source. In conclusion, results of this study suggest that binding of IgG1 and IgG2 by neutrophils is unchanged or increased while that for IgM is decreased after migration into the mammary gland. Furthermore, in vitro phagocytosis of Staphylococcus aureus is enhanced after migration.
Comparative Clinical Pathology – Springer Journals
Published: Oct 7, 2004
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