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Published ahead of print on March 6, 2003, doi:10.1165/rcmb.2002-0275OC

Am. J. Respir. Cell Mol. Biol., Volume 29, Number 2, August 2003, 239-244

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Submitted on November 26, 2002
Revised on February 28, 2003

Peroxynitrite Enhances Interleukin-10 Reduction in the Release of Neutrophil Chemotactic Activity

Hiroki Numanami1, Dan K Nelson2, Jeffrey C Hoyt3, Jon L Freels3, Michael P Habib3, Jun Amano4, Masayuki Haniuda4, Sekiya Koyama5, and Richard A Robbins3*

1 Research Service, Southern Arizona VA Health Care System, Tucson, AZ, USA; Arizona Respiratory Center, University of Arizona, Tucson, AZ, USA; Second Department of Surgery, Shinshu University, Matsumoto, Japan, 2 Research Service, Southern Arizona VA Health Care System, Tucson, AZ, USA, 3 Research Service, Southern Arizona VA Health Care System, Tucson, AZ, USA; Arizona Respiratory Center, University of Arizona, Tucson, AZ, USA, 4 Second Department of Surgery, Shinshu University, Matsumoto, Japan, 5 First Department of Internal Medicine, Shinshu University, Matsumoto, Japan

* To whom correspondence should be addressed. E-mail: Richard.Robbins2{at}med.va.gov.

Peroxynitrite, formed by nitric oxide and superoxide, has been shown to nitrate and reduce the function of pro-inflammatory proteins such as interleukin (IL)-8, monocyte chemoattractant protein-1, and eotaxin, but in contrast, enhance the function of the anti-inflammatory cytokine, IL-10, in reducing IL-1 release from blood monocytes. However, the effect of nitrated IL-10 on release of pro-inflammatory cytokines from lung epithelial cells is unknown. We hypothesized that peroxynitrite would enhance the capacity of human IL-10 to reduce inflammatory mediators released by epithelial cells. To test this hypothesis, recombinant human IL-10 was evaluated for its capacity to attenuate the release of neutrophil chemotactic activity (NCA) and IL-8 from a human epithelial cell line in response to IL-1{beta} and tumor necrosis factor {alpha} (TNF{alpha}). NCA and IL-8 in lung epithelial culture supernatant fluids were significantly lower after culture with nitrated human IL-10 compared to non-nitrated human IL-10 controls (p<0.05). Consistent with these results, nitrated human IL-10 attenuated IL-8 mRNA expression more than non-nitrated human IL-10 controls (p<0.05). These data demonstrate that peroxynitrite exposed IL-10 has enhanced anti-inflammatory activity and suggest that nitration may play a critical role in the regulation of inflammation within the lower respiratory tract.






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