RA Robbins, JH Sisson, DR Springall, KJ Nelson, JA Taylor, NA Mason, JM Polak and RG Townley
Research Service, Overton Brooks VA Medical Center, Shreveport, Louisiana 71101, USA.

Nitric oxide (NO) is a gas released by the airway epithelium, but the mechanism regulating NO release is unclear. We hypothesized that lung mononuclear cell release of tumor necrosis factor alpha (TNF) and interleukin-1beta (IL-1) would induce epithelial cells to release NO. Lung mononuclear cells were obtained from seven normal volunteers by bronchoalveolar lavage and cultured with Escherichia coli lipopolysaccharide for 24 h. The mononuclear cell culture-conditioned media (M-CM) were then applied to cultures of the murine lung epithelial cell line, LA-4. Nitrite and nitrite + nitrate concentrations were 0.9 +/- 0.1 and 11.8 +/- 2.4 microM in the M-CM. Culturing LA-4 cells line with the M-CM (1:10 dilution) resulted in a marked and time-dependent increase in nitrite or nitrite + nitrate compared with LA-4 cells cultured in media alone (2.4 +/- 0.5 versus 0.9 +/- 0.1 microm and 16.6 +/- 0.6 versus 11.8 +/- 2.4 microM after 24 h). Antibodies to TNF and/or IL-1 significantly reduced the nitrite or nitrite + nitrate concentrations and the concentrations of TNF and IL-1 in the M-CM correlated with nitrite concentrations in the LA-4 culture supernatant fluids (r2 = 0.848 and 0.956). Inducible nitric oxide synthase (iNOS) protein and mRNA examined by immunohistochemistry and Northern blot analysis revealed a marked elevation in the cells cultured with the M-CM which was significantly reduced by TNF and IL-1 antibodies. These data demonstrate that mononuclear cells can stimulate LA-4 cells to express iNOS by releasing TNF and IL-1.

This article has been cited by other articles:

				Y. Yan, J. Li, W. Ouyang, Q. Ma, Y. Hu, D. Zhang, J. Ding, Q. Qu, K. Subbaramaiah, and C. Huang NFAT3 is specifically required for TNF-{alpha}-induced cyclooxygenase-2 (COX-2) expression and transformation of Cl41 cells J. Cell Sci., July 15, 2006; 119(14): 2985 - 2994. [Abstract] [Full Text] [PDF]

				J. C. Hoyt, J. Ballering, H. Numanami, J. M. Hayden, and R. A. Robbins Doxycycline Modulates Nitric Oxide Production in Murine Lung Epithelial Cells J. Immunol., January 1, 2006; 176(1): 567 - 572. [Abstract] [Full Text] [PDF]

				I. Gertsberg, V. Hellman, M. Fainshtein, S. Weil, S. D. Silberberg, M. Danilenko, and Z. Priel Intracellular Ca2+ Regulates the Phosphorylation and the Dephosphorylation of Ciliary Proteins Via the NO Pathway J. Gen. Physiol., October 25, 2004; 124(5): 527 - 540. [Abstract] [Full Text] [PDF]

				K. Subbaramaiah, K. Yoshimatsu, E. Scherl, K. M. Das, K. D. Glazier, D. Golijanin, R. A. Soslow, T. Tanabe, H. Naraba, and A. J. Dannenberg Microsomal Prostaglandin E Synthase-1 Is Overexpressed in Inflammatory Bowel Disease: EVIDENCE FOR INVOLVEMENT OF THE TRANSCRIPTION FACTOR Egr-1 J. Biol. Chem., March 26, 2004; 279(13): 12647 - 12658. [Abstract] [Full Text] [PDF]

				D.V. Pechkovsky, G. Zissel, C. Stamme, T. Goldmann, H. Ari Jaffe, M. Einhaus, C. Taube, H. Magnussen, M. Schlaak, and J. Muller-Quernheim Human alveolar epithelial cells induce nitric oxide synthase-2 expression in alveolar macrophages Eur. Respir. J., April 1, 2002; 19(4): 672 - 683. [Abstract] [Full Text] [PDF]

				A. Murakami, K. Kawabata, T. Koshiba, G. Gao, Y. Nakamura, K. Koshimizu, and H. Ohigashi Nitric Oxide Synthase Is Induced in Tumor Promoter-sensitive, but not Tumor Promoter-resistant, JB6 Mouse Epidermal Cells Cocultured with Interferon- {{gamma}}-stimulated RAW 264.7 Cells: The Role of Tumor Necrosis Factor-{{alpha}} Cancer Res., November 1, 2000; 60(22): 6326 - 6331. [Abstract] [Full Text]

				B. M. Fischer, L. G. Rochelle, J. A. Voynow, N. J. Akley, and K. B. Adler Tumor Necrosis Factor-alpha Stimulates Mucin Secretion and Cyclic GMP Production by Guinea Pig Tracheal Epithelial Cells In Vitro Am. J. Respir. Cell Mol. Biol., March 1, 1999; 20(3): 413 - 422. [Abstract] [Full Text]

				K. L. Jones, T. W. Bryan, P. A. Jinkins, K. L. Simpson, M. B. Grisham, M. W. Owens, S. A. Milligan, B. A. Markewitz, and R. A. Robbins Superoxide released from neutrophils causes a reduction in nitric oxide gas Am J Physiol Lung Cell Mol Physiol, December 1, 1998; 275(6): L1120 - L1126. [Abstract] [Full Text] [PDF]

				R. A. Robbins, P. A. Jinkins, T. W. Bryan, S. C. Prado, and S. A. Milligan Methotrexate Inhibition of Inducible Nitric Oxide Synthase in Murine Lung Epithelial Cells In Vitro Am. J. Respir. Cell Mol. Biol., June 1, 1998; 18(6): 853 - 859. [Abstract] [Full Text]

				D. V. Pechkovsky, G. Zissel, T. Goldmann, M. Einhaus, C. Taube, H. Magnussen, M. Schlaak, and J. Muller-Quernheim Pattern of NOS2 and NOS3 mRNA expression in human A549 cells and primary cultured AEC II Am J Physiol Lung Cell Mol Physiol, April 1, 2002; 282(4): L684 - L692. [Abstract] [Full Text] [PDF]