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Construction of novel in vitro epithelioid cell granuloma model from mouse macrophage cell line

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Abstract

There have been several attempts to make granuloma model to clarify the mechanism of granulomatous diseases like sarcoidosis. However, a unique in vitro model that generates multinucleated giant cell (MGC) through epithelioid cells resembled to human granuloma, has not yet been clearly established. In this study, the generation of granuloma model that forms MGC via epithelioid cells from the mouse macrophage cell line was investigated. A RAW 246.7 mouse macrophage cell line was cultured with lipopolysaccharide (LPS) and concanavalin A (Con A) in various concentrations either alone or both. We found that separate treatment of LPS and Con A induced around 35 and 20% MGC respectively whereas cotreatment of these chemicals drastically accelerated granuloma formation rate and it was around 80%. The highest fusion index (MGC formation rate) was observed at days 7. A gradual increase of tumor necrosis factor alpha (TNF-α) production in the culture supernatant was analyzed by enzyme-linked immunosorbent assay (ELISA). And the neutralization of the elevated level of TNF-α production by its monoclonal antibody leads to significant decrease of MGC formation. Interestingly, we found that the RAW cells were changed into spindle cells, which morphologically resembled to epithelioid cells and eventually MGC was formed from these spindle cells. Our in vitro granuloma model appeared to be similar with in vivo epithelioid cell granulomas like sarcoidosis. Thus, our model would be useful as in vitro epithelioid granuloma model for analyzing the mechanisms and screening the effective drugs of granulomatous diseases in future.

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References

  1. Byrd TF (1998) Multinucleated giant cell formation induced by IFN-γ/IL-3 is associated with restriction of virulent mycobacterium tuberculosis cell to cell invasion in human monocytes monolayers. Cell Immunol 188:89–96

    Article  PubMed  CAS  Google Scholar 

  2. Graber R, Leoni L, Carrel S, Losa GA (1993) Lectin and anti-T monoclonal antibodies-induced changes of second messengers generating enzymes in human peripheral blood mononuclear cells. Cell Mol Biol 39:45–54

    PubMed  CAS  Google Scholar 

  3. Held TK, Weihua X, Yuan L, Kalvakolanu DV, Cross AS (1999) Gamma interferon augments macrophage activation by lipopolysaccharide by two distinct mechanisms, at the signal transduction level and via an autocrine mechanism involving tumor necrosis factor alpha and interleukin-1. Infect Immun 67:206–212

    PubMed  CAS  Google Scholar 

  4. Ikeda T, Ikeda K, Sasaki K, Kawakami K, Hatake K, Kaji Y, Norimatsu H, Harada M, Takahara J (1998) IL-13 as well as IL-4 induces monocytes/macraphages and a monoblastic cell line (UG3) to differentiate into multinucleated giant cells in the presense of M-CSF. Biochem Biophys Res Commun 253:265–272

    Article  PubMed  CAS  Google Scholar 

  5. Jersmann HP, Hii CS, Ferrante JV, Ferrante A (2001) Bacterial lipopolysaccharide and tumor necrosis factor alpha synergistically increase expression of human endothelial adhesion molecules thought activation of NF-κB and p38 mitogen-activated protein kinase signaling pathways. Infect Immun 69:1273–1279

    Article  PubMed  CAS  Google Scholar 

  6. Mallbris L, Ljungberg A, Hedblad MA, Larsson P, Stahle-Backdahl M (2003) Progressive cutaneous sarcoidosis responding to anti-tumor necrosis factor-α therapy. J Am Acad Dermatol 48:290–293

    Article  PubMed  Google Scholar 

  7. Mcinnes A, Rennick DM (1988) Interleukin 4 induces cultured monocytes -macrophages to form giant multinucleated cells. J Exp Med 167:598–611

    Article  PubMed  CAS  Google Scholar 

  8. McNally AK, Anderson JM (1995) Interleukin-4 induce foreign body giant cells from human monocytes/macrophages. Differential lymphokine regulation of macrophage fusion leads to morphological variants of multinucleated giant cells. Am J Pathol 147:1487–1499

    PubMed  CAS  Google Scholar 

  9. Mitchell DN, Scadding JG, Heard BE. Hinson KF (1977) Sarcoidosis: histopathological definition and clinical diagnosis. J Clin Pathol 30:395–408

    Article  PubMed  CAS  Google Scholar 

  10. Mizuno K, Okamoto H, Horio T (2004) Inhibitory influences of xanthine oxidase inhibitor and angiotensin I-converting enzyme inhibitor on multinucleated giant cell formation from monocytes by downregulation of adhesion molecules and purinergic receptors. Br J Dermatol 150:205–210

    Article  PubMed  CAS  Google Scholar 

  11. Morrison DC, Ryan JL (1979) Bacterial endotoxins and host immune responses. Adv Immunol 28:293–450

    Article  PubMed  CAS  Google Scholar 

  12. Munoz-Fernandez MA, Pimentel-Muinos FX, Alonso MA, Campanero M, Sanchez-Madrid F, Silva A, Alonso JL, Fresno M (1990) Synergy of tumor necrosis factor with protein kinase C activators on T cell activation. Eur J Immunol 20:605–610

    Article  PubMed  CAS  Google Scholar 

  13. Okamoto H, Mizuno K, Horio T (2003) Monocyte-derived multinucleated giant cells and sarcoidosis. J Dermatol Sci 31:119–128

    Article  PubMed  CAS  Google Scholar 

  14. Puissegur MP, Lay G, Gilleron M, Botella L, Nigou J, Marrakchi H, Mari B, Duteyrat JL, Guerardel Y, Kremer L, Barbry P, Puzo G, Altare F (2007) Mycobacterial lipomannan induces granuloma macrophage fusion via a TLR2-dependent, ADAM9- and β1 integrin-mediated pathway. J Immunol 178:3161–3169

    PubMed  CAS  Google Scholar 

  15. Sone S (1984) Function of multinucleated giant cells formed by fusing rat alveolar macrophage with lymphokines containing macrophages fusion factor. Lymphokine Res; 3:163–173

    PubMed  CAS  Google Scholar 

  16. Suen YK, Fung KP, Choy YM, Lee CY, Chan CW, Kong SK (2000) Concanavalin A induced apoptosis in murine macrophage PU5-1.8 cells through clustering of mitochondria and release of cytochrome c. Apoptosis 5:369–377

    Article  PubMed  CAS  Google Scholar 

  17. Zhao R, Guerrah A, Tang H, Zhao ZJ (2002) Cell surface glycoprotein PZR is a major mediator of concanabalin A-induced cell signaling. J Bio Chem 277:7882–7888

    Article  CAS  Google Scholar 

  18. Zughaier SM, Zimmer SM, Datta A, Carlson RW, Stephens DS (2005) Differential induction of the Toll-like receptor 4-MyD88-dependent and-independent signaling pathways by endotoxins. Infect Immun 73:2940–2950

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank A. Morikawa for excellent technical assistance and T. Yokochi (Department of Microbiology and Immunology, Aichi Medical University School of Medicine) for his helpful suggestion during the course of this study.

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Authors and Affiliations

  1. Department of Dermatology, Aichi Medical University School of Medicine, Nagakute, Aichi, 480-1195, Japan

    Takeshi Yanagishita, Daisuke Watanabe, Yoichi Akita, Aki Nakano, Yuichiro Ohshima, Yasuhiko Tamada & Yoshinari Matsumoto

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  1. Takeshi Yanagishita
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  2. Daisuke Watanabe
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  3. Yoichi Akita
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  4. Aki Nakano
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  5. Yuichiro Ohshima
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  6. Yasuhiko Tamada
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  7. Yoshinari Matsumoto
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Correspondence to Takeshi Yanagishita.

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Yanagishita, T., Watanabe, D., Akita, Y. et al. Construction of novel in vitro epithelioid cell granuloma model from mouse macrophage cell line. Arch Dermatol Res 299, 399–403 (2007). https://doi.org/10.1007/s00403-007-0778-1

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  • DOI: https://doi.org/10.1007/s00403-007-0778-1

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