Human lung mononuclear cells induce nitric oxide synthase in murine airway epithelial cells in vitro

Role of TNFα and IL-1β

Richard A. Robbins, Joseph H. Sisson, David R. Springall, Kenneth J. Nelson, Janice A. Taylor, Nicola A. Mason, Julia M. Polak, Robert G. Townley

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

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 α (TNF) and interleukin- 1β3 (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 μM 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 μm and 16.6 ± 0.6 versus 11.8 ± 2.4 μM 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.

Original languageEnglish
Pages (from-to)268-273
Number of pages6
JournalAmerican Journal of Respiratory and Critical Care Medicine
Volume155
Issue number1
StatePublished - 1997
Externally publishedYes

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Interleukin-13
Nitrites
Nitric Oxide Synthase
Tumor Necrosis Factor-alpha
Epithelial Cells
Conditioned Culture Medium
Culture Media
Lung
Cell Culture Techniques
Nitrates
Nitric Oxide
Nitric Oxide Synthase Type II
Cultured Cells
Cell Line
Antibodies
Bronchoalveolar Lavage
In Vitro Techniques
Northern Blotting
Lipopolysaccharides
Healthy Volunteers

All Science Journal Classification (ASJC) codes

  • Pulmonary and Respiratory Medicine

Cite this

Robbins, R. A., Sisson, J. H., Springall, D. R., Nelson, K. J., Taylor, J. A., Mason, N. A., ... Townley, R. G. (1997). Human lung mononuclear cells induce nitric oxide synthase in murine airway epithelial cells in vitro: Role of TNFα and IL-1β. American Journal of Respiratory and Critical Care Medicine, 155(1), 268-273.

Human lung mononuclear cells induce nitric oxide synthase in murine airway epithelial cells in vitro : Role of TNFα and IL-1β. / Robbins, Richard A.; Sisson, Joseph H.; Springall, David R.; Nelson, Kenneth J.; Taylor, Janice A.; Mason, Nicola A.; Polak, Julia M.; Townley, Robert G.

In: American Journal of Respiratory and Critical Care Medicine, Vol. 155, No. 1, 1997, p. 268-273.

Research output: Contribution to journalArticle

Robbins, RA, Sisson, JH, Springall, DR, Nelson, KJ, Taylor, JA, Mason, NA, Polak, JM & Townley, RG 1997, 'Human lung mononuclear cells induce nitric oxide synthase in murine airway epithelial cells in vitro: Role of TNFα and IL-1β', American Journal of Respiratory and Critical Care Medicine, vol. 155, no. 1, pp. 268-273.
Robbins, Richard A. ; Sisson, Joseph H. ; Springall, David R. ; Nelson, Kenneth J. ; Taylor, Janice A. ; Mason, Nicola A. ; Polak, Julia M. ; Townley, Robert G. / Human lung mononuclear cells induce nitric oxide synthase in murine airway epithelial cells in vitro : Role of TNFα and IL-1β. In: American Journal of Respiratory and Critical Care Medicine. 1997 ; Vol. 155, No. 1. pp. 268-273.
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