Cooperative effect of tumor necrosis factor and gamma-interferon on
chemotactic peptide receptor expression and stimulus-induced actin
polymerization in HL-60 cells
KM Rao, MA Misukonis, HJ Cohen and JB Weinberg
Geriatric Research, Education and Clinical Center, Veterans Administration
Medical Center, Durham, NC 27705.
We studied the effect of tumor necrosis factor (TNF) and gamma interferon
(IFN-gamma), alone and in combination, on the expression of chemotactic
peptide receptors, stimulus-induced actin polymerization, hydrogen peroxide
production (H2O2), and expression of nonspecific esterase (NSE) positivity
in human promyelocytic leukemic cell line HL- 60. These parameters were
analyzed following a five-day culture with the cytokines. Chemotactic
peptide receptor expression was studied using the fluoresceinated
hexapeptide, formyl-norleucyl-leucyl- phenylalanyl-norleucyl-tyrosyl-lysine
and flow cytometry. Actin polymerization, an important event required for
chemotaxis and phagocytosis, was studied using NBD-phallacidin labeling,
following stimulation with the chemotactic peptide formyl-methionyl-leucyl-
phenylalanine (FMLP) or phorbol myristate acetate (PMA). TNF increased the
expression of chemotactic peptide receptors in a dose-dependent fashion,
and there was good correlation between the receptor expression,
stimulus-induced actin polymerization, H2O2 production, and NSE positivity.
IFN-gamma was less potent in inducing all the parameters studied but
exerted a positive cooperative effect when combined with TNF. IFN-gamma at
high concentrations induced chemotactic peptide receptors comparable in
magnitude to that seen with TNF but failed to prime these cells to undergo
actin polymerization in response to FMLP or PMA. Undifferentiated HL-60
cells showed a decrease in F- actin content on stimulation with PMA. This
suggests that protein kinase C might have a negative regulatory role in
stimulus-induced actin polymerization. The observations reported here
indicate that appropriate combinations of different inducing agents with
different modes of action might be necessary to duplicate the functional
abilities of mature phagocytic cells.
Volume 71,
Issue 4,
pp. 1062-1067,
04/01/1988
Copyright © 1988 by The American Society of Hematology