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Notch signaling regulates expression of Mcl-1 and apoptosis in PPD-treated macrophages

Cellular & Molecular Immunology volume 10, pages 444–452 (2013)Cite this article

Abstract

Macrophages are cellular targets for infection by bacteria and viruses. The fate of infected macrophages plays a key role in determining the outcome of the host immune response. Apoptotic cell death of macrophages is considered to be a protective host defense that eliminates pathogens and infected cells. In this study, we investigated the involvement of Notch signaling in regulating apoptosis in macrophages treated with tuberculin purified protein derivative (PPD). Murine bone marrow-derived macrophages (BMMs) treated with PPD or infected with Mycobacterium bovis Bacillus Calmette-Guérin (BCG) induced upregulation of Notch1. This upregulation correlated well with the upregulation of the anti-apoptotic gene mcl-1 both at the transcriptional and translational levels. Decreased levels of Notch1 and Mcl-1 were observed in BMM treated with PPD when a gamma secretase inhibitor (GSI), which inhibits the processing of Notch receptors, was used. Moreover, silencing Notch1 in the macrophage-like cell line RAW264.7 decreased Mcl-1 protein expression, suggesting that Notch1 is critical for Mcl-1 expression in macrophages. A significant increase in apoptotic cells was observed upon treatment of BMM with PPD in the presence of GSI compared to the vehicle-control treated cells. Finally, analysis of the mcl-1 promoter in humans and mice revealed a conserved potential CSL/RBP-Jκ binding site. The association of Notch1 with the mcl-1 promoter was confirmed by chromatin immunoprecipitation. Taken together, these results indicate that Notch1 inhibits apoptosis of macrophages stimulated with PPD by directly controlling the mcl-1 promoter.

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Acknowledgements

The authors are grateful to Dr. Stefan H. E. Kaufmann, Dr Barbara Osborne and Dr Todd Golde for sharing reagents. The authors thank Dr Jorg Schriber and Dr Masami Nakatsu for their help with ChIP and ChIP-on-chip analysis. This work was partly supported by the Thailand Research Fund Grant No. RSA5280014, Research Foundation Enhancement from Chulalongkorn University (Ratchadaphiseksomphot Endowment Fund) and by the Thai Government Stimulus Package 2 (TKK2555) under the Project for Establishment of Comprehensive Center for Innovative Food, Health Products and Agriculture and the Higher Education Research Promotion and National Research University Project of Thailand, the Office of the Higher Education Commission (AS613A) and The Fogarty International Research Collaborative Award (Grant No. R03 TW008420-01A1, NIH, Bethesda, MD USA). W. Wongchana was supported by the Thailand Research Fund through the Royal Golden Jubilee Ph.D. Program (PHD/0337/2551) (NIH, Bethesda, MD, USA).

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Tanapat Palaga, Siriluk Ratanabunyong, Thitiporn Pattarakankul, Naunpun Sangphech, Wipawee Wongchana & Patipark Kueanjinda

  2. Interdisciplinary Program in Medical Microbiology, Graduate School, Chulalongkorn University, Bangkok, 10330, Thailand

    Siriluk Ratanabunyong, Naunpun Sangphech & Patipark Kueanjinda

  3. Graduate Program in Industrial Microbiology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Thitiporn Pattarakankul

  4. Graduate Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Wipawee Wongchana

  5. International Center for Biotechnology, Osaka University, Osaka, 565-0871, Japan

    Yukihiro Hadae

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Correspondence to Tanapat Palaga.

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Palaga, T., Ratanabunyong, S., Pattarakankul, T. et al. Notch signaling regulates expression of Mcl-1 and apoptosis in PPD-treated macrophages. Cell Mol Immunol 10, 444–452 (2013). https://doi.org/10.1038/cmi.2013.22

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