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ComparativeIn VitroPhagocytosis and F‐Actin Polymerization of Bovine Neonatal Neutrophils*

F. Moiola, M. Spycher, Marianne Wyder-Walther, R Zwahlen

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Abstract

Analysis of neonatal neutrophil (PMN) functions should help to reveal factors which could contribute to the impaired host defense system of neonates. We analysed functional parameters of PMN from newborn calves (N-PMN) and adult bovines (A-PMN): cellular volume and F-actin content upon stimulation with complement factors, by cytofluorometry and phagocytosis of E. coli 78:80B with a colorimetric assay. Polymerization of F-actin was rapid in both N- and A-PMN, but reached higher levels in N-PMN. N-PMN are significantly smaller than A-PMN throughout the whole activation time. Percentage of phagocytosing PMN, the rate of phagocytosis, and the rate of killing are similar between A- and N-PMN after opsonization of bacteria with adult serum (AS). Opsonization with newborn serum (NS) reduced all three examined parameters: in A-PMN more (P < 0.001) than in N-PMN (P < 0.05). However, when phagocytosis was compared following age-matched opsonization (N-PMN and NS; A-PMN and AS), N-PMN phagocytosed less (P < 0.001) bacteria per PMN than A-PMN. Additionally, steroidal (dexamethasone) and non-steroidal (phenylbutazone) anti-inflammatory drugs inhibited phagocytosis by N-PMN less than by A-PMN. Higher relative F-actin content of N-PMN can be correlated with the documented functional hyperactivity of bovine N-PMN. However, the exaggerated impairment of phagocytosis in calves observed after age-matched opsonization of bacteria could potentially indicate a specific host defence defect.

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What this paper is about

Analysis of neonatal neutrophil (PMN) functions should help to reveal factors which could contribute to the impaired host defense system of neonates. We analysed functional parameters of PMN from newborn calves (N-PMN) and adult bovines (A-PMN): cellular volume and F-actin content upon stimulation with complement factors, by cytofluorometry and phagocytosis of E. coli 78:80B with a colorimetric assay. Polymerization of F-actin was rapid in both N- and A-PMN, but reached higher levels in N-PMN. N-PMN are significantly smaller than A-PMN throughout the whole activation time. Percentage of phagocytosing PMN, the rate of phagocytosis, and the rate of killing are similar between A- and N-PMN after opsonization of bacteria with adult serum (AS). Opsonization with newborn serum (NS) reduced all three examined parameters: in A-PMN more (P < 0.001) than in N-PMN (P < 0.05). However, when phagocytosis was compared following age-matched opsonization (N-PMN and NS; A-PMN and AS), N-PMN phagocytosed less (P < 0.001) bacteria per PMN than A-PMN. Additionally, steroidal (dexamethasone) and non-steroidal (phenylbutazone) anti-inflammatory drugs inhibited phagocytosis by N-PMN less than by A-PMN. Higher relative F-actin content of N-PMN can be correlated with the documented functional hyperactivity of bovine N-PMN. However, the exaggerated impairment of phagocytosis in calves observed after age-matched opsonization of bacteria could potentially indicate a specific host defence defect.

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Available abstract

Analysis of neonatal neutrophil (PMN) functions should help to reveal factors which could contribute to the impaired host defense system of neonates. We analysed functional parameters of PMN from newborn calves (N-PMN) and adult bovines (A-PMN): cellular volume and F-actin content upon stimulation with complement factors, by cytofluorometry and phagocytosis of E. coli 78:80B with a colorimetric assay. Polymerization of F-actin was rapid in both N- and A-PMN, but reached higher levels in N-PMN. N-PMN are significantly smaller than A-PMN throughout the whole activation time. Percentage of phagocytosing PMN, the rate of phagocytosis, and the rate of killing are similar between A- and N-PMN after opsonization of bacteria with adult serum (AS). Opsonization with newborn serum (NS) reduced all three examined parameters: in A-PMN more (P < 0.001) than in N-PMN (P < 0.05). However, when phagocytosis was compared following age-matched opsonization (N-PMN and NS; A-PMN and AS), N-PMN phagocytosed less (P < 0.001) bacteria per PMN than A-PMN. Additionally, steroidal (dexamethasone) and non-steroidal (phenylbutazone) anti-inflammatory drugs inhibited phagocytosis by N-PMN less than by A-PMN. Higher relative F-actin content of N-PMN can be correlated with the documented functional hyperactivity of bovine N-PMN. However, the exaggerated impairment of phagocytosis in calves observed after age-matched opsonization of bacteria could potentially indicate a specific host defence defect.

Key concepts: Antibody opsonization, Phagocytosis, Microbiology, In vitro, Granulocyte, Immunology, Biology, Bacteria

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