Membrane NADPH oxidase activity and cell size in bovine neonatal and adult neutrophils.

Abstract:

:Neutrophils (PMN) from newborn calves generate significantly less superoxide anion (O2-) than do their adult counterparts after stimulation with direct protein kinase C agonists. To better understand this observation, we compared the activity and kinetics of NADPH oxidase in membrane fractions from phorbol 12-myristate 13-acetate-stimulated adult and newborn PMN. After phorbol 12-myristate 13-acetate stimulation, PMN were sonicated and the membranes assayed for O2- production with increasing concentrations of NADPH. O2- production was calculated 1 and 2 min after the beginning of the reaction. At all concentrations of NADPH used, there was no difference (p greater than 0.05) in O2- production between adult (n = 8) and newborn (n = 9) PMN membrane preparations. Enzyme kinetics calculations revealed no differences (p greater than 0.05) between age groups in Km and Vmax or in the velocity of the reactions. Determination of the protein content in the membrane pellet, however, showed that adult PMN yielded significantly (p less than 0.01) higher amounts of protein (2.82 +/- 0.14 mg/mL) than did newborn PMN (1.78 +/- 0.07 mg/mL). This difference could be partly attributed to cell size; flow cytometric analysis showed that newborn PMN had a significantly (p less than 0.01) smaller diameter (10.94 +/- 0.07 microns) than did adult PMN (11.65 +/- 0.06 microns), and calculated cell volume and surface area were also both significantly less (p less than 0.01) in newborn PMN.(ABSTRACT TRUNCATED AT 250 WORDS)

journal_name

Pediatr Res

journal_title

Pediatric research

authors

Doré M,Slauson DO,Neilsen NR

doi

10.1203/00006450-199010000-00005

subject

Has Abstract

pub_date

1990-10-01 00:00:00

pages

327-31

issue

4

eissn

0031-3998

issn

1530-0447

journal_volume

28

pub_type

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