Infectious and autoimmune diseases may promote sleep by down-regulating circadian gene expression
By Melissa Lee Phillips | July 17, 2007
The immune system alters mammalian circadian clock gene expression, perhaps explaining why fatigue accompanies some infections and autoimmune diseases, according to a study in this week's Proceedings of the National Academy of Sciences. The authors found that a protein involved in mammalian immunity interferes with circadian clock gene expression and promotes sleep in mice.
"The study certainly provides a framework within which to address the question of how immunomodulators affect both the timing and the quantity or quality of sleep," said Mark Opp of the University of Michigan in Ann Arbor, who was not involved in the study.
The immune response to microbial infection activates pro-inflammatory molecules such as tumor necrosis factor alpha (TNF-α). Previous work has suggested that TNF-α causes lethargy and fatigue in people with cancer, rheumatoid arthritis, and sleep apnea. Animal studies have also shown that TNF-α enhances sleep, but the mechanism has remained unknown.
To see if TNF-α directly regulates circadian oscillations, researchers led by Gionata Cavadini of University Hospital Zurich in Switzerland analyzed TNF-α's effects on expression of circadian clock genes.
"There's very little known about how [immune response] is related to circadian rhythms," said James Krueger of Washington State University in Pullman, who was not involved in the work.
The central circadian oscillator in mammals lies in the suprachiasmatic nucleus (SCN) of the hypothalamus, but other body cells also contain peripheral oscillators that keep their own rhythms. Interacting loops of gene transcription and translation form a negative feedback system that generates the 24-hour mammalian clock.
The researchers first found that, in culture, TNF-α suppresses the expression of three circadian clock genes called Period, and the expression of three transcription factors they controlled by clock genes. TNF-α only affected the genes' expression levels; it did not affect the times at which they were expressed.
Other work has shown that activation of Period genes as well as the transcription factors depends on a specific DNA sequence - called an E-box - lying upstream of these genes. Transcription factors bind to the E-box sequence to initiate these genes' transcription.
When Cavadini and her colleagues mutated the E-box sequence, administering TNF-α did not suppress transcription. Conversely, TNF-α did not activate circadian clock genes whose expression does not depend on an E-box sequence.
Their results suggest that TNF-α affects only clock genes that contain an E-box and that the reduced activity of these genes affects sleep regulation, co-author Thomas Birchler, also of University Hospital Zurich, told The Scientist in an Email.
When the researchers tested the effects of TNF-α in live mice, they found that animals treated with the cytokine kept normal cycles -- sleeping during the day and becoming active at night - but they moved less and rested more during their active periods than did untreated mice. These results paralleled in vitro findings showing that TNF-α changes clock genes' expression levels but not timing, the authors say. Also, as in the in vitro studies, TNF-α down-regulated expression of Period and the three clock-controlled transcription factors in the liver.
It's possible that compensating effects by other clock-related genes is keeping the circadian period normal in mice treated with TNF-α, senior author Adriano Fontana of University Hospital Zurich told The Scientist in an Email. "This may explain why TNF-α, which influences the expression of several clock genes, does not result in a shift of the circadian clock."
Another possibility, however, is that TNF-α is affecting not the central circadian pacemaker in the SCN, but only peripheral oscillators in the liver or other tissues, according to Vincent Cassone of Texas A&M University in College Station, who wasn't involved in the study. If Period gene expression is suppressed, "you should get a change in [circadian] period," he said. Since the authors don't see that, Cassone suspects that they've found "purely a direct effect on output and peripheral oscillations, which I think is kind of interesting."
The authors believe that TNF-α is affecting SCN rhythms, because they found a slight reduction of one of the clock-controlled transcription factors in the SCN of TNF-α-treated mice. However, they didn't look at expression of the other clock genes.
According to Krueger, the study's findings fit in well with previous findings about immune response effects on sleep. However, he said, "I don't think TNF's effects on sleep are entirely dependent on activating or deactivating the circadian rhythm genes." TNF-α also controls expression of many other genes, Krueger said, and some of these likely influence sleep in other ways.
From an evolutionary point of view, increased sleep during illness may help animals to fight off infections or may keep sick animals from socializing and spreading an infection to others, Fontana said. It's possible that increased sleep is merely a byproduct of illness, Opp said, but many researchers believe that "the way we sleep when we are sick facilitates recovery."
Melissa Lee Phillips
Links within this article
K.Y. Kreeger, "Collecting clues to the mammalian clock," The Scientist, April 15, 2002.
G. Cavadini et al., "TNF-α suppresses the expression of clock genes by
interfering with E-box-mediated transcription," PNAS, published online July 16, 2007.
J. Lucentini, "The body sleeps, but the genes do not," The Scientist, February 16, 2004.
S. Akira et al., "Pathogen recognition and innate immunity," Cell, February 24, 2006.
D.R. Spriggs et al., "Recombinant human tumor necrosis factor administered as a 24-hour intravenous infusion. A phase I and pharmacologic study," Journal of the National Cancer Institute, September 7, 1988.
L.C. Pollard et al., "Fatigue in rheumatoid arthritis reflects pain, not disease activity," Rheumatology (Oxford), July 2006.
A.N. Vgontzas et al., "Marked decrease in sleepiness in patients with sleep apnea by etanercept, a tumor necrosis factor-alpha antagonist," Journal of Clinical Endocrinology and Metabolism, September 2004. Sep;89(9):4409-13.
J.M. Krueger, J.A. Majde, "Humoral links between sleep and the immune system: research issues," Annals of the New York Academy of Sciences, May 2003.
J.P. Roberts, "What sets the biological clock?" The Scientist, June 10, 2002.
U. Schibler, P. Sassone-Corsi, "A web of circadian pacemakers," Cell, December 27, 2002.
N. Cermakian, P. Sassone-Corsi, "Multilevel regulation of the circadian clock," Nature Reviews Molecular and Cell Biology, October 2000.