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. 2002 Jan;110(1):79–84. doi: 10.1289/ehp.0211079

The effect of ozone exposure on the ability of human surfactant protein a variants to stimulate cytokine production.

Guirong Wang 1, Todd M Umstead 1, David S Phelps 1, Hamid Al-Mondhiry 1, Joanna Floros 1
PMCID: PMC1240696  PMID: 11781168

Abstract

Ozone exposure can cause inflammation and impaired lung function. Human surfactant protein A (SP-A) may play a role in inflammation by modulating cytokine production by macrophages. SP-A is encoded by two genes, SP-A1 and SP-A2, and several allelic variants have been characterized for each gene. These allelic variants differ among themselves in amino acids that may exhibit differential sensitivity to ozone-induced oxidation and this may produce functional differences. We studied the effects of SP-A variants before and after ozone exposure on the production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-8. These are important proinflammatory cytokines and are expressed by the macrophage-like THP-1 cells. Eight variants were expressed in vitro, characterized by gel electrophoresis, and studied. These included six single-gene SP-A alleles and two SP-A variants derived from both genes. Variants were exposed to ozone at 1 ppm for 4 hr at 37 degrees C, and we compared their ability to stimulate cytokine (TNF-alpha and IL-8) production by THP-1 cells to air-exposed and unexposed SP-A variants. We found that a) SP-A2 variants (1A, 1A(0), 1A(1) stimulate significantly more TNF-alpha and IL-8 production than SP-A1 variants (6A, 6A(2), 6A(4); b) coexpressed SP-A variants (1A(0)/6A(2), 1A(1)/6A(4) have significantly higher activity than single gene products; c) after ozone exposure, all SP-A variants showed a decreased ability to stimulate TNF-alpha and IL-8 production, and the level of the decrease varied among SP-A variants (26-48%); and d) human SP-A from patients with alveolar proteinosis exhibited a minimal decrease (18% and 12%, respectively) in its ability to stimulate TNF-alpha and IL-8 after in vitro ozone exposure. We conclude that biochemical and functional differences exist among SP-A variants, that ozone exposure modulates the ability of SP-A variants to stimulate cytokines by THP-1 cells, and that SP-As from bronchoalveolar lavage (BAL) fluid of certain alveolar proteinosis patients may be oxidized in vivo.

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Selected References

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  1. Balis J. U., Paterson J. F., Haller E. M., Shelley S. A., Montgomery M. R. Ozone-induced lamellar body responses in a rat model for alveolar injury and repair. Am J Pathol. 1988 Aug;132(2):330–344. [PMC free article] [PubMed] [Google Scholar]
  2. Balis J. U., Paterson J. F., Lundh J. M., Haller E. M., Shelley S. A., Montgomery M. R. Ozone stress initiates acute perturbations of secreted surfactant membranes. Am J Pathol. 1991 Apr;138(4):847–857. [PMC free article] [PubMed] [Google Scholar]
  3. Batenburg J. J., Haagsman H. P. The lipids of pulmonary surfactant: dynamics and interactions with proteins. Prog Lipid Res. 1998 Sep;37(4):235–276. doi: 10.1016/s0163-7827(98)00011-3. [DOI] [PubMed] [Google Scholar]
  4. Buchman A. R., Berg P. Comparison of intron-dependent and intron-independent gene expression. Mol Cell Biol. 1988 Oct;8(10):4395–4405. doi: 10.1128/mcb.8.10.4395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crouch E., Chang D., Rust K., Persson A., Heuser J. Recombinant pulmonary surfactant protein D. Post-translational modification and molecular assembly. J Biol Chem. 1994 Jun 3;269(22):15808–15813. [PubMed] [Google Scholar]
  6. Devlin R. B., McDonnell W. F., Mann R., Becker S., House D. E., Schreinemachers D., Koren H. S. Exposure of humans to ambient levels of ozone for 6.6 hours causes cellular and biochemical changes in the lung. Am J Respir Cell Mol Biol. 1991 Jan;4(1):72–81. doi: 10.1165/ajrcmb/4.1.72. [DOI] [PubMed] [Google Scholar]
  7. DiAngelo S., Lin Z., Wang G., Phillips S., Ramet M., Luo J., Floros J. Novel, non-radioactive, simple and multiplex PCR-cRFLP methods for genotyping human SP-A and SP-D marker alleles. Dis Markers. 1999 Dec;15(4):269–281. doi: 10.1155/1999/961430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Floros J., Hoover R. R. Genetics of the hydrophilic surfactant proteins A and D. Biochim Biophys Acta. 1998 Nov 19;1408(2-3):312–322. doi: 10.1016/s0925-4439(98)00077-5. [DOI] [PubMed] [Google Scholar]
  9. Floros J., Phelps D. S., Taeusch H. W. Biosynthesis and in vitro translation of the major surfactant-associated protein from human lung. J Biol Chem. 1985 Jan 10;260(1):495–500. [PubMed] [Google Scholar]
  10. Fornstedt N., Porath J. Characterization studies on a new lectin found in seeds of Vicia ervilia. FEBS Lett. 1975 Sep 15;57(2):187–191. doi: 10.1016/0014-5793(75)80713-7. [DOI] [PubMed] [Google Scholar]
  11. Horstman D. H., Folinsbee L. J., Ives P. J., Abdul-Salaam S., McDonnell W. F. Ozone concentration and pulmonary response relationships for 6.6-hour exposures with five hours of moderate exercise to 0.08, 0.10, and 0.12 ppm. Am Rev Respir Dis. 1990 Nov;142(5):1158–1163. doi: 10.1164/ajrccm/142.5.1158. [DOI] [PubMed] [Google Scholar]
  12. Johansson J., Curstedt T. Molecular structures and interactions of pulmonary surfactant components. Eur J Biochem. 1997 Mar 15;244(3):675–693. doi: 10.1111/j.1432-1033.1997.00675.x. [DOI] [PubMed] [Google Scholar]
  13. Kremlev S. G., Phelps D. S. Surfactant protein A stimulation of inflammatory cytokine and immunoglobulin production. Am J Physiol. 1994 Dec;267(6 Pt 1):L712–L719. doi: 10.1152/ajplung.1994.267.6.L712. [DOI] [PubMed] [Google Scholar]
  14. Kremlev S. G., Umstead T. M., Phelps D. S. Surfactant protein A regulates cytokine production in the monocytic cell line THP-1. Am J Physiol. 1997 May;272(5 Pt 1):L996–1004. doi: 10.1152/ajplung.1997.272.5.L996. [DOI] [PubMed] [Google Scholar]
  15. Kulle T. J., Sauder L. R., Hebel J. R., Chatham M. D. Ozone response relationships in healthy nonsmokers. Am Rev Respir Dis. 1985 Jul;132(1):36–41. doi: 10.1164/arrd.1985.132.1.36. [DOI] [PubMed] [Google Scholar]
  16. Malhotra R., Haurum J., Thiel S., Sim R. B. Interaction of C1q receptor with lung surfactant protein A. Eur J Immunol. 1992 Jun;22(6):1437–1445. doi: 10.1002/eji.1830220616. [DOI] [PubMed] [Google Scholar]
  17. Mason R. J., Greene K., Voelker D. R. Surfactant protein A and surfactant protein D in health and disease. Am J Physiol. 1998 Jul;275(1 Pt 1):L1–13. doi: 10.1152/ajplung.1998.275.1.L1. [DOI] [PubMed] [Google Scholar]
  18. McCormack F. X., Calvert H. M., Watson P. A., Smith D. L., Mason R. J., Voelker D. R. The structure and function of surfactant protein A. Hydroxyproline- and carbohydrate-deficient mutant proteins. J Biol Chem. 1994 Feb 25;269(8):5833–5841. [PubMed] [Google Scholar]
  19. McCormack F. The structure and function of surfactant protein-A. Chest. 1997 Jun;111(6 Suppl):114S–119S. doi: 10.1378/chest.111.6_supplement.114s. [DOI] [PubMed] [Google Scholar]
  20. Mudd J. B., Leavitt R., Ongun A., McManus T. T. Reaction of ozone with amino acids and proteins. Atmos Environ. 1969 Nov;3(6):669–682. doi: 10.1016/0004-6981(69)90024-9. [DOI] [PubMed] [Google Scholar]
  21. Phelps D. S., Floros J., Taeusch H. W., Jr Post-translational modification of the major human surfactant-associated proteins. Biochem J. 1986 Jul 15;237(2):373–377. doi: 10.1042/bj2370373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Putman E., van Golde L. M., Haagsman H. P. Toxic oxidant species and their impact on the pulmonary surfactant system. Lung. 1997;175(2):75–103. doi: 10.1007/pl00007561. [DOI] [PubMed] [Google Scholar]
  23. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  24. Wang G., Phelps D. S., Umstead T. M., Floros J. Human SP-A protein variants derived from one or both genes stimulate TNF-alpha production in the THP-1 cell line. Am J Physiol Lung Cell Mol Physiol. 2000 May;278(5):L946–L954. doi: 10.1152/ajplung.2000.278.5.L946. [DOI] [PubMed] [Google Scholar]

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