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Regulation of Cytokine mRNA Expression in Lipopolysaccharide-Stimulated Human Macrophages

W. William Zhong, MD, MSc; Peter A. Burke, MD; A. Teresa Hand, MD, FRCS; Mark J. Walsh, MS; Laura A. Hughes; R. Armour Forse, MD, PhD
Arch Surg. 1993;128(2):158-164. doi:10.1001/archsurg.1993.01420140035006.
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• One of the responses of the human macrophage to lipopolysaccharide (LPS) is the production of a number of cytokines. The regulation of these cytokines is still not clearly understood. To study this regulation, mRNA levels of interleukin 1α (IL-1α), IL-1β, tumor necrosis factor alpha (TNF-α), IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and IL-8/neutrophil chemotactic factor were determined in 10-day-old differentiated macrophages following stimulation with a low dose of LPS (0.001 to 10 ng/mL) with use of the polymerase chain reaction. Increased levels of mRNA for IL-8 were detectable after exposure to a very low dose of LPS (0.001 ng/mL) and levels of IL-1 β and TNF-α were detectable only after stimulation with doses of 0.01 ng/mL. The mRNA for IL-8 was detected 30 minutes after the addition of LPS, while those for IL-1 β and TNF-α were only measurable at 1 hour. The mRNAs for IL-1α, IL-6, and GM-CSF were detectable only with a higher dose of lipopolysaccharide and only after a longer exposure time. In addition, the messages for IL-6 and GM-CSF were measurable for a short time, while those of IL-8 and of IL-1 β were detectable for a longer time. The secretion of TNF-α and GM-CSF tightly followed gene activation, and that of IL-6 and IL-8 steadily increased even after the mRNA level of these cytokines returned to baseline. Secretion of IL-1α and IL-1 β was hardly detected, although their gene activation was obvious. These data indicate that cytokine mRNA levels following lipopolysaccharide stimulation are highly regulated. Individual cytokines show variable patterns of response. These responses are both dose and time dependent and are not necessarily associated with the secretion of protein.

(Arch Surg. 1993;128:158-164)


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