Regulation of cytokine mRNA expression in lipopolysaccharide-stimulated human macrophages

W. W. Zhong, P. A. Burke, A. T. Hand, M. J. Walsh, L. A. Hughes and R. A. Forse
Department of Surgery, New England Deaconess Hospital, Harvard Medical School, Boston, Mass 02215.

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 1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor alpha (TNF-alpha), 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 beta and TNF-alpha 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 beta and TNF-alpha were only measurable at 1 hour. The mRNAs for IL-1 alpha, 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 beta were detectable for a longer time. The secretion of TNF-alpha 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-1 alpha and IL-1 beta 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.

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES

Anti-B7-1/B7-2 antibody elicits innate-effector responses in macrophages through NF-{kappa}B-dependent pathway
Khan et al.
Int Immunol 2007;19:477-486.
ABSTRACT | FULL TEXT  

TLR4-mediated inflammatory activation of human coronary artery endothelial cells by LPS
Zeuke et al.
Cardiovasc Res 2002;56:126-134.
ABSTRACT | FULL TEXT  

Macrophage Effector Functions Controlled by Bruton's Tyrosine Kinase Are More Crucial Than the Cytokine Balance of T Cell Responses for Microfilarial Clearance
Mukhopadhyay et al.
J. Immunol. 2002;168:2914-2921.
ABSTRACT | FULL TEXT  

Comparison of SP-A and LPS effects on the THP-1 monocytic cell line
Song and Phelps
Am. J. Physiol. Lung Cell. Mol. Physiol. 2000;279:L110-L117.
ABSTRACT | FULL TEXT  

Detection of Granulocyte-Macrophage Colony-Stimulating Factor in Patients with Pulmonary Alveolar Proteinosis
CARRAWAY et al.
Am. J. Respir. Crit. Care Med. 2000;161:1294-1299.
ABSTRACT | FULL TEXT  

Regulation of Periodontal Ligament Cell Functions by Interleukin-1beta
Agarwal et al.
Infect. Immun. 1998;66:932-937.
ABSTRACT | FULL TEXT  

Lipopolysaccharide activation of human monocytes mediated by CD14, results in a coordinated synthesis of tissue factor, TNF-{alpha} and IL-6
Osnes et al.
Innate Immunity 1995;2:27-35.
ABSTRACT