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STAT1-dependent and STAT1-independent gene expression in murine immune cells following stimulation with interferon-alpha

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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Purpose

The precise molecular targets of interferon-alpha (IFN-α) therapy of melanoma are unknown but likely involve signal transducer and activator of transcription 1 (STAT1) signal transduction within host immune effector cells. We hypothesized that microarray analysis could be utilized to identify candidate molecular targets important for mediating the anti-tumor effect of exogenously administered IFN-α.

Experimental Methods

To identify the STAT1-dependent genes regulated by IFN-α, the gene expression profile of splenocytes from wild type (WT) and STAT1−/− mice was characterized.

Results

This analysis identified 30 genes that required STAT1 signal transduction for optimal expression in response to IFN-α (p < 0.001). These genes include granzyme b (Gzmb), interferon regulatory factor 7 (Irf7), Fas death domain-associated protein (Daxx), and lymphocyte antigen 6 complex, locus C (Ly6c). The expression of 20 genes was found to be suppressed in the presence of STAT1 including chemokine ligand 2 (Ccl2), Ccl5, and Ccl7. Nineteen genes were significantly upregulated in murine splenocytes following treatment with IFN-α regardless of the presence of STAT1 including CD86, lymphocyte antigen 6 complex, locus A (Ly6a), and Tap binding protein (Tapbp). The expression of representative IFN-responsive genes was confirmed at the transcriptional level by Real Time PCR.

Conclusion

This report is the first to demonstrate that STAT1-mediated signal transduction plays a major role in the transcriptional response of murine immune cells to IFNα.

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Acknowledgments

The authors thank The Ohio State University Comprehensive Cancer Center (CCC) Microarray Core Facility for performing cRNA hybridization and the raw data collection. We also thank The Ohio State University CCC Real Time Core Facility for assisting in the operation of the ABI PRISM 7900 Sequence Detection System. Some statistical analyses were performed using BRB-ArrayTools Version 3.22 developed by Dr. Richard Simon and Amy Peng Lam. This work was supported by National Institutes of Health (NIH) Grants K24 CA93670, P30-CA16058 (to WE Carson) and a Career Development Award from the Melanoma Research Foundation (to GB Lesinski).

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Authors and Affiliations

  1. Integrated Biological Sciences Graduate Program, The Ohio State University Comprehensive Cancer Center, Columbus, OH, 43210, USA

    Jason M. Zimmerer

  2. Human Cancer Genetics Program Department of Molecular Virology, Immunology, and Medical Genetics, The Ohio State University Comprehensive Cancer Center, Columbus, OH, 43210, USA

    Jason M. Zimmerer, Gregory B. Lesinski & William E. Carson III

  3. Center for Biostastistics, The Ohio State University Comprehensive Cancer Center, Columbus, OH, 43210, USA

    Michael D. Radmacher & Amy Ruppert

  4. Division of Surgical Oncology, Department of Surgery, The Ohio State University Comprehensive Cancer Center, N924 Doan Hall, 410 West 10th Ave., Columbus, OH, 43210, USA

    William E. Carson III

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  1. Jason M. Zimmerer
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  2. Gregory B. Lesinski
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  3. Michael D. Radmacher
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  4. Amy Ruppert
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  5. William E. Carson III
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Correspondence to William E. Carson III.

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Zimmerer, J.M., Lesinski, G.B., Radmacher, M.D. et al. STAT1-dependent and STAT1-independent gene expression in murine immune cells following stimulation with interferon-alpha. Cancer Immunol Immunother 56, 1845–1852 (2007). https://doi.org/10.1007/s00262-007-0329-9

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  • DOI: https://doi.org/10.1007/s00262-007-0329-9

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