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Cytokine gene expression profile in monocytic cells after a co-culture with epithelial cells

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Abstract

Epithelial cells represent an important source of cytokines that may modulate the influx and functions of mononuclear phagocytes. The aim of our study was to characterize changes in the gene expression of selected cytokines in human macrophages co-cultured with respiratory epithelial cells. The A549 alveolar type II-like cell line was co-cultured with THP-1 cells (monocyte/macrophage cell line) in filter-separated mode to avoid their cell–cell contact. At different time-points (0, 4, 8, 12 and 24 h), the cells were harvested separately to evaluate their gene and protein expression (IL-1 beta, IL-6, IL-8, IL-10 and GM-CSF). Quantitative RT-PCR analysis showed prominent changes in the THP-1 cytokine gene expression induced by a co-culture with A549 cells. Fourfold upregulation of mRNA expression has been found in 12 genes and 4-fold downregulation in 5 genes as compared to the unstimulated control sample with a p value smaller than 0.05. The induction of inhibin beta A and IL-1 beta mRNA after 12 h and the expression of IL-1 alpha and GM-CSF mRNA after 24 h were the most prominent. When looking at the cytokine levels in culture supernatants, IL-1 beta and IL-8 were induced early (at 8 h) as compared to the release of IL-6 and GM-CSF (at 24 h). We conclude that respiratory epithelial cells constitutively regulate the cytokine gene expression of macrophages located in their environment and might further modulate the release of cytokines by posttranslational pathways.

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References

  1. Gordon S, Taylor PR. Monocyte and macrophage heterogeneity. Nat Rev Immunol. 2005;5(12):953–64.

    Article  PubMed  CAS  Google Scholar 

  2. Mills CD, Kincaid K, Alt JM, Heilman MJ, Hill AM. M-1/M-2 macrophages and the Th1/Th2 paradigm. J Immunol. 2000;164(12):6166–73.

    PubMed  CAS  Google Scholar 

  3. Kuroda E, Kito T, Yamashita U. Reduced expression of STAT4 and IFN-gamma in macrophages from BALB/c mice. J Immunol. 2002;168(11):5477–82.

    PubMed  CAS  Google Scholar 

  4. Verreck FA, de Boer T, Langenberg DM, Hoeve MA, Kramer M, Vaisberg E, et al. Human IL-23-producing type 1 macrophages promote but IL-10-producing type 2 macrophages subvert immunity to (myco) bacteria. Proc Natl Acad Sci USA. 2004;101(13):4560–5.

    Article  PubMed  CAS  Google Scholar 

  5. Goerdt S, Orfanos CE. Other functions, other genes: alternative activation of antigen-presenting cells. Immunity. 1999;10(2):137–42.

    Article  PubMed  CAS  Google Scholar 

  6. Gough MJ, Melcher AA, Ahmed A, Crittenden MR, Riddle DS, Linardakis E, et al. Macrophages orchestrate the immune response to tumor cell death. Cancer Res. 2001;61(19):7240–7.

    PubMed  CAS  Google Scholar 

  7. Gordon S. Alternative activation of macrophages. Nat Rev Immunol. 2003;3(1):23–35.

    Article  PubMed  CAS  Google Scholar 

  8. Buhling F, Ittenson A, Kaiser D, Tholert G, Hoffmann B, Reinhold D, et al. MRP8/MRP14, CD11b and HLA-DR expression of alveolar macrophages in pneumonia. Immunol Lett. 2000;71(3):185–90.

    Article  PubMed  CAS  Google Scholar 

  9. Leonard CT, Soccal PM, Singer L, Berry GJ, Theodore J, Holt PG, et al. Dendritic cells and macrophages in lung allografts: a role in chronic rejection? Am J Respir Crit Care Med. 2000;161(4 Pt 1):1349–54.

    PubMed  CAS  Google Scholar 

  10. Hamilton JA. GM-CSF in inflammation and autoimmunity. Trends Immunol. 2002;23(8):403–8.

    Article  PubMed  CAS  Google Scholar 

  11. Stanley E, Lieschke GJ, Grail D, Metcalf D, Hodgson G, Gall JA, et al. Granulocyte/macrophage colony-stimulating factor-deficient mice show no major perturbation of hematopoiesis but develop a characteristic pulmonary pathology. Proc Natl Acad Sci USA. 1994;91(12):5592–6.

    Article  PubMed  CAS  Google Scholar 

  12. Striz I, Slavcev A, Kalanin J, Jaresova M, Rennard SI. Cell-cell contacts with epithelial cells modulate the phenotype of human macrophages. Inflammation. 2001;25(4):241–6.

    Article  PubMed  CAS  Google Scholar 

  13. Sims JE, Smith DE. The IL-1 family: regulators of immunity. Nat Rev Immunol. 2010;10(2):89–102.

    Article  PubMed  CAS  Google Scholar 

  14. Brunda MJ. Interleukin-12. J Leukoc Biol. 1994;55(2):280–8.

    PubMed  CAS  Google Scholar 

  15. D’Andrea A, Rengaraju M, Valiante NM, Chehimi J, Kubin M, Aste M, et al. Production of natural killer cell stimulatory factor (interleukin 12) by peripheral blood mononuclear cells. J Exp Med. 1992;176(5):1387–98.

    Article  PubMed  Google Scholar 

  16. Eramaa M, Hurme M, Stenman UH, Ritvos O. Activin A/erythroid differentiation factor is induced during human monocyte activation. J Exp Med. 1992;176(5):1449–52.

    Article  PubMed  CAS  Google Scholar 

  17. Eto Y, Tsuji T, Takezawa M, Takano S, Yokogawa Y, Shibai H. Purification and characterization of erythroid differentiation factor (EDF) isolated from human leukemia cell line THP-1. Biochem Biophys Res Commun. 1987;142(3):1095–103.

    Article  PubMed  CAS  Google Scholar 

  18. Shao L, Frigon NL Jr, Sehy DW, Yu AL, Lofgren J, Schwall R, et al. Regulation of production of activin A in human marrow stromal cells and monocytes. Exp Hematol. 1992;20(10):1235–42.

    PubMed  CAS  Google Scholar 

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Acknowledgments

This study was supported by the MSMT grant ME 906 and by the Institute for Clinical and Experimental Medicine (MZO 00023001).

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Authors and Affiliations

  1. Department of Immunogenetics, Institute for Clinical and Experimental Medicine, Videnska 1958/9, Prague, Czech Republic

    Libor Kolesar

  2. Department of Clinical and Transplant Immunology, Institute for Clinical and Experimental Medicine, Videnska 1958/9, Prague, Czech Republic

    Libor Kolesar, Eva Brabcova, Eliska Thorburn, Alena Sekerkova, Marcela Jaresova & Ilja Striz

  3. Transplant Laboratory, Institute for Clinical and Experimental Medicine, Videnska 1958/9, Prague, Czech Republic

    Irena Brabcova

  4. Department of Nephrology, Institute for Clinical and Experimental Medicine, Videnska 1958/9, Prague, Czech Republic

    Ondrej Viklicky

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  1. Libor Kolesar
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  2. Eva Brabcova
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  3. Eliska Thorburn
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  5. Irena Brabcova
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  8. Ilja Striz
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Correspondence to Libor Kolesar.

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Kolesar, L., Brabcova, E., Thorburn, E. et al. Cytokine gene expression profile in monocytic cells after a co-culture with epithelial cells. Immunol Res 52, 269–275 (2012). https://doi.org/10.1007/s12026-012-8338-y

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  • DOI: https://doi.org/10.1007/s12026-012-8338-y

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