Cancer Immunology, Immunotherapy

, Volume 56, Issue 11, pp 1845–1852

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

  • Jason M. Zimmerer
  • Gregory B. Lesinski
  • Michael D. Radmacher
  • Amy Ruppert
  • William E. CarsonIII
Original Article

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.

Keywords

Interferon-alpha Oligonucleotide microarray analysis STAT1 Immune cells 

References

  1. 1.
    Badgwell B, Lesinski GB, Magro C, Abood G, Skaf A, Carson W III (2004) The antitumor effects of interferon-alpha are maintained in mice challenged with a STAT1-deficient murine melanoma cell line. J Surg Res 116:129–136PubMedCrossRefGoogle Scholar
  2. 2.
    Bauer JA, Morrison BH, Grane RW, Jacobs BS, Borden EC, Lindner DJ (2003) IFN-alpha2b and thalidomide synergistically inhibit tumor-induced angiogenesis. J Interferon Cytokine Res 23:3–10PubMedCrossRefGoogle Scholar
  3. 3.
    Belardelli F, Ferrantini M, Proietti E, Kirkwood JM (2002) Interferon-alpha in tumor immunity and immunotherapy. Cytokine Growth Factor Rev 13:119–134PubMedCrossRefGoogle Scholar
  4. 4.
    Belardelli F, Gresser I, Maury C, Maunoury MT (1982) Antitumor effects of interferon in mice injected with interferon-sensitive and interferon-resistant Friend leukemia cells. I. Int J Cancer 30:813–820CrossRefGoogle Scholar
  5. 5.
    Belardelli F, Gresser I, Maury C, Maunoury MT (1982) Antitumor effects of interferon in mice injected with interferon-sensitive and interferon-resistant Friend leukemia cells. II. Role of host mechanisms. Int J Cancer 30:821–825PubMedCrossRefGoogle Scholar
  6. 6.
    Biron CA (2001) Interferons alpha and beta as immune regulators–a new look. Immunity 14:661–664PubMedCrossRefGoogle Scholar
  7. 7.
    Brassard DL, Grace MJ, Bordens RW (2002) Interferon-alpha as an immunotherapeutic protein. J Leukoc Biol 71:565–581PubMedGoogle Scholar
  8. 8.
    Carr DJ, Chodosh J, Ash J, Lane TE (2003) Effect of anti-CXCL10 monoclonal antibody on herpes simplex virus type 1 keratitis and retinal infection. J Virol 77:10037–10046PubMedCrossRefGoogle Scholar
  9. 9.
    Darnell JE Jr, Kerr IM, Stark GR (1994) Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins. Science 264:1415–1421PubMedCrossRefGoogle Scholar
  10. 10.
    Durbin JE, Hackenmiller R, Simon MC, Levy DE (1996) Targeted disruption of the mouse Stat1 gene results in compromised innate immunity to viral disease. Cell 84:443–450PubMedCrossRefGoogle Scholar
  11. 11.
    Fallarino F, Gajewski TF (1999) Cutting edge: differentiation of antitumor CTL in vivo requires host expression of Stat1. J Immunol 163:4109–4113PubMedGoogle Scholar
  12. 12.
    Frey AB (2006) Myeloid suppressor cells regulate the adaptive immune response to cancer. J Clin Invest 116:2587–2590PubMedCrossRefGoogle Scholar
  13. 13.
    Fuse S, Obar JJ, Bellfy S, Leung EK, Zhang W, Usherwood EJ (2006) CD80 and CD86 control antiviral CD8+ T-cell function and immune surveillance of murine gammaherpesvirus 68. J Virol 80:9159–9170PubMedCrossRefGoogle Scholar
  14. 14.
    Garbe C, Krasagakis K, Zouboulis CC, Schroder K, Kruger S, Stadler R, Orfanos CE (1990) Antitumor activities of interferon alpha, beta, and gamma and their combinations on human melanoma cells in vitro: changes of proliferation, melanin synthesis, and immunophenotype. J Invest Dermatol 95:231S–237SPubMedCrossRefGoogle Scholar
  15. 15.
    Gil MP, Bohn E, O’Guin AK, Ramana CV, Levine B, Stark GR, Virgin HW, Schreiber RD (2001) Biologic consequences of Stat1-independent IFN signaling. Proc Natl Acad Sci USA 98:6680–6685PubMedCrossRefGoogle Scholar
  16. 16.
    Grant EP, Michalek MT, Goldberg AL, Rock KL (1995) Rate of antigen degradation by the ubiquitin-proteasome pathway influences MHC class I presentation. J Immunol 155:3750–3758PubMedGoogle Scholar
  17. 17.
    Gresser I, Kaido T, Maury C, Woodrow D, Moss J, Belardelli F (1994) Interaction of IFN alpha/beta with host cells essential to the early inhibition of Friend erythroleukemia visceral metastases in mice. Int J Cancer 57:604–611PubMedCrossRefGoogle Scholar
  18. 18.
    Gresser I, Maury C, Carnaud C, De Maeyer E, Maunoury MT, Belardelli F (1990) Anti-tumor effects of interferon in mice injected with interferon-sensitive and interferon-resistant Friend erythroleukemia cells. VIII. Role of the immune system in the inhibition of visceral metastases. Int J Cancer 46:468–474PubMedCrossRefGoogle Scholar
  19. 19.
    Haque SJ, Williams BR (1998) Signal transduction in the interferon system. Semin Oncol 25:14–22PubMedGoogle Scholar
  20. 20.
    Imanishi T, Kamigaki T, Nakamura T, Hayashi S, Yasuda T, Kawasaki K, Takase S, Ajiki T, Kuroda Y (2006) Correlation between expression of major histocompatibility complex class I and that of antigen presenting machineries in carcinoma cell lines of the pancreas, biliary tract and colon. Kobe J Med Sci 52:85–95PubMedGoogle Scholar
  21. 21.
    Jaakkola I, Merinen M, Jalkanen S, Hanninen A (2003) Ly6C induces clustering of LFA-1 (CD11a/CD18) and is involved in subtype-specific adhesion of CD8 T cells. J Immunol 170:1283–1290PubMedGoogle Scholar
  22. 22.
    Koon H, Atkins M (2006) Autoimmunity and immunotherapy for cancer. N Engl J Med 354:758–760PubMedCrossRefGoogle Scholar
  23. 23.
    Lesinski GB, Anghelina M, Zimmerer J, Bakalakos T, Badgwell B, Parihar R, Hu Y, Becknell B, Abood G, Chaudhury AR, Magro C, Durbin J, Carson WE III (2003) The antitumor effects of IFN-alpha are abrogated in a STAT1-deficient mouse. J Clin Invest 112:170–180PubMedCrossRefGoogle Scholar
  24. 24.
    Lesinski GB, Kondadasula SV, Crespin T, Shen L, Kendra K, Walker M, Carson WE III (2004) Multiparametric flow cytometric analysis of inter-patient variation in STAT1 phosphorylation following interferon Alfa immunotherapy. J Natl Cancer Inst 96:1331–1342PubMedCrossRefGoogle Scholar
  25. 25.
    Levy D, Reich N, Kessler D, Pine R, Darnell JE Jr (1988) Transcriptional regulation of interferon-stimulated genes: a DNA response element and induced proteins that recognize it. Cold Spring Harb Symp Quant Biol 53(Pt 2):799–802PubMedGoogle Scholar
  26. 26.
    Li C, Wong WH (2003) The analysis of gene expression data: methods and software. DNA-Chip Analyzer (dChip)Google Scholar
  27. 27.
    Lin MY, Zal T, Ch’en IL, Gascoigne NR, Hedrick SM (2005) A pivotal role for the multifunctional calcium/calmodulin-dependent protein kinase II in T cells: from activation to unresponsiveness. J Immunol 174:5583–5592PubMedGoogle Scholar
  28. 28.
    Maellaro E, Pacenti L, Del Bello B, Valentini MA, Mangiavacchi P, De Felice C, Rubegni P, Luzi P, Miracco C (2003) Different effects of interferon-alpha on melanoma cell lines: a study on telomerase reverse transcriptase, telomerase activity and apoptosis. Br J Dermatol 148:1115–1124PubMedCrossRefGoogle Scholar
  29. 29.
    Moschella F, Bisikirska B, Maffei A, Papadopoulos KP, Skerrett D, Liu Z, Hesdorffer CS, Harris PE (2003) Gene expression profiling and functional activity of human dendritic cells induced with IFN-alpha-2b: implications for cancer immunotherapy. Clin Cancer Res 9:2022–2031PubMedGoogle Scholar
  30. 30.
    Moschos SJ, Edington HD, Land SR, Rao UN, Jukic D, Shipe-Spotloe J, Kirkwood JM (2006) Neoadjuvant treatment of regional stage IIIB melanoma with high-dose interferon alfa-2b induces objective tumor regression in association with modulation of tumor infiltrating host cellular immune responses. J Clin Oncol 24:3164–3171PubMedCrossRefGoogle Scholar
  31. 31.
    Mullbacher A, Waring P, Tha Hla R, Tran T, Chin S, Stehle T, Museteanu C, Simon MM (1999) Granzymes are the essential downstream effector molecules for the control of primary virus infections by cytolytic leukocytes. Proc Natl Acad Sci USA 96:13950–13955PubMedCrossRefGoogle Scholar
  32. 32.
    Parronchi P, Mohapatra S, Sampognaro S, Giannarini L, Wahn U, Chong P, Maggi E, Renz H, Romagnani S (1996) Effects of interferon-alpha on cytokine profile, T cell receptor repertoire and peptide reactivity of human allergen-specific T cells. Eur J Immunol 26:697–703PubMedCrossRefGoogle Scholar
  33. 33.
    Ramana CV, Gil MP, Han Y, Ransohoff RM, Schreiber RD, Stark GR (2001) Stat1-independent regulation of gene expression in response to IFN-gamma. Proc Natl Acad Sci USA 98:6674–6679PubMedCrossRefGoogle Scholar
  34. 34.
    Rizvi SM, Raghavan M (2006) Direct peptide-regulatable interactions between MHC class I molecules and tapasin. Proc Natl Acad Sci USA 103:18220–18225PubMedCrossRefGoogle Scholar
  35. 35.
    Saborit-Villarroya I, Del Valle JM, Romero X, Esplugues E, Lauzurica P, Engel P, Martin M (2005) The adaptor protein 3BP2 binds human CD244 and links this receptor to Vav signaling, ERK activation, and NK cell killing. J Immunol 175:4226–4235PubMedGoogle Scholar
  36. 36.
    Zimmerer JM, Lesinski GB, Kondadasula SV, Karpa VI, Lehman A, Raychaudhury A, Becknell B, Carson WE III (2007) IFN-{alpha}-Induced signal transduction, gene expression, and antitumor activity of immune effector cells are negatively regulated by suppressor of cytokine signaling proteins. J Immunol 178:4832–4845PubMedGoogle Scholar
  37. 37.
    Zimmerer JM, Lesinski GB, Kornacker K, Shen L, Liyanarachichi S, Durbin J, Kendra K, Walker M, Carson WE (2003) Gene expression profiling of the response to interferon-alpha immunotherapy. American Association for Cancer Research Annual Meeting, Orlando, FL [Abstract no. 4679]Google Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Jason M. Zimmerer
    • 1
    • 2
  • Gregory B. Lesinski
    • 2
  • Michael D. Radmacher
    • 3
  • Amy Ruppert
    • 3
  • William E. CarsonIII
    • 2
    • 4
  1. 1.Integrated Biological Sciences Graduate ProgramThe Ohio State University Comprehensive Cancer CenterColumbusUSA
  2. 2.Human Cancer Genetics Program Department of Molecular Virology, Immunology, and Medical GeneticsThe Ohio State University Comprehensive Cancer CenterColumbusUSA
  3. 3.Center for BiostastisticsThe Ohio State University Comprehensive Cancer CenterColumbusUSA
  4. 4.Division of Surgical Oncology, Department of SurgeryThe Ohio State University Comprehensive Cancer CenterColumbusUSA

Personalised recommendations