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Department of Respiratory, Allergy, Immunology, Inflammation, and Infectious Diseases, Pfizer Central Research, Groton, CT 06340
Cultured monocytes and macrophages stimulated with LPS produce
large quantities of proIL-1ß, but release little mature cytokine to
the medium. The efficiency at which the procytokine is converted to its
active 17-kDa species and released extracellularly is enhanced by
treating cytokine-producing cells with a secretion stimulus such as ATP
or nigericin. To determine whether this need for a secretion stimulus
extends to blood, individual donors were bled twice daily for 4
consecutive days, and the collected blood samples were subjected to a
two-step IL-1 production assay. LPS-activated blood samples generated
cell-free IL-1ß, but levels of the extracellular cytokine were
greatly increased by subsequent treatment with ATP or nigericin.
Specificity and concentration requirements of the nucleotide
triphosphate effect suggests a P2X7 receptor involvement.
Quantities of IL-1ß generated by an individual donor’s blood in
response to the LPS-only and LPS/ATP stimuli were relatively consistent
over the 4-day period. Between donors, consistent differences in
cytokine production capacity were observed. Blood samples treated with
ATP also demonstrated enhanced IL-18 production, but TNF-
levels
decreased. Among leukocytes, monocytes appeared to be the most affected
cellular targets of the ATP stimulus. These studies indicate that an
exogenous stimulus is required by blood for the efficient production of
IL-1ß and IL-18, and suggest that circulating blood monocytes
constitutively express a P2X7-like
receptor.
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