DIFFERENTIAL IN-VITRO EFFECTS OF PHYSIOLOGICAL AND ATMOSPHERIC OXYGEN-TENSION ON NORMAL HUMAN PERIPHERAL-BLOOD MONONUCLEAR CELL-PROLIFERATION, CYTOKINE AND IMMUNOGLOBULIN PRODUCTION
Ja. Krieger et al., DIFFERENTIAL IN-VITRO EFFECTS OF PHYSIOLOGICAL AND ATMOSPHERIC OXYGEN-TENSION ON NORMAL HUMAN PERIPHERAL-BLOOD MONONUCLEAR CELL-PROLIFERATION, CYTOKINE AND IMMUNOGLOBULIN PRODUCTION, International journal of immunopharmacology, 18(10), 1996, pp. 545-552
Since the early work of Mischell & Dutton (Science 153, 1004-1008, 196
6), it has been recognized that certain lymphocyte cultures are exquis
itely sensitive to the harsh effects of atmospheric oxygen tension. Th
e influence of oxygen partial pressure (pO(2)) on normal human periphe
ral blood mononuclear cell (PBMC) phenotype, proliferative ability, cy
tokine, immunoglobulin production, and redox status was examined by cu
lturing PBMC under ambient oxygen (high pO(2)) or a more physiological
pO(2) (5% O-2; low pO(2)). Low pO(2) conditions promoted a significan
t increase in overall viable PBMC number and enhanced Concanavalin A (
Con A)- or pokeweed mitogen (PWM)-stimulated PBMC proliferation by app
roximately 30% and 50%, respectively. No differential pO(2) effects we
re apparent on phytohemagglutinin (PHA)- or staphylococcal enterotoxin
B (SEB)-induced proliferation. Both resting and Con A-stimulated lymp
hocytes incubated for 24 h under high pO(2) had a greater baseline car
boxy-2'7'-dichlorofluorescin (C-DCF) fluorescence, and were less able
to quench the effect of H2O2 treatment compared to lymphocytes culture
d under low pO(2) conditions. Supernatant gamma-IFN, IL-2, and IL-4 co
ncentrations were elevated 50-65% when PBMC were stimulated with Con A
for 24 h under low pO(2); however, lipopolysaccharide (LPS)-stimulate
d IL-1 beta production was reduced by over 75%. PWM-stimulated IgM pro
duction by PBMC was significantly reduced in day 7 cultures incubated
under low pO(2), whereas IgG and IgA production remained relatively un
altered. Immunophenotyping analyses did not reveal any significant alt
erations in cell subset or marker distribution at the time points exam
ined; however, an interesting trend of increased CD69 expression was o
bserved for Con A-stimulated PBMC incubated under low pO(2). These res
ults demonstrate that O-2 isa critical parameter for the in vitro cult
ure of lymphocytes, and suggests that varying pO(2) may differentially
alter PBMC functionality. (C) 1997 International Society for Immunoph
armacology.