Skip to main content
SpringerLink
Log in

Retroviral Transduction and Reporter Assay: Transcription Factor Cooperation in Th9 Cell Development

  • Protocol
  • First Online:
Th9 Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1585))

  • 1234 Accesses

Abstract

Naïve CD4+ T cells differentiate into different T helper subsets in response to specific cytokine environment and transcription factors. Th9 cells are induced in response to signals from cytokines, TGF-β and IL-4. Transcription factors that are downstream of these cytokines converge to drive the development of Th9 cells. Retroviral transduction allows the genetic modification in T cells thereby helping us to better understand the molecular mechanisms that control their development as well as function. In this chapter, an optimized protocol for retroviral transduction of murine Th9 cells as well as transient transfection of Th9 cells with luciferase reporter constructs is described.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 119.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 159.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Zhu J, Yamane H, Paul WE (2010) Differentiation of effector CD4 T cell populations (*). Annu Rev Immunol 28:445–489. doi:10.1146/annurev-immunol-030409-101212

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Zhou L, Chong MM, Littman DR (2009) Plasticity of CD4+ T cell lineage differentiation. Immunity 30(5):646–655. doi:10.1016/j.immuni.2009.05.001

    Article  CAS  PubMed  Google Scholar 

  3. Veldhoen M, Uyttenhove C, van Snick J, Helmby H, Westendorf A, Buer J, Martin B, Wilhelm C, Stockinger B (2008) Transforming growth factor-beta ‘reprograms’ the differentiation of T helper 2 cells and promotes an interleukin 9-producing subset. Nat Immunol 9(12):1341–1346. doi:10.1038/ni.1659

    Article  CAS  PubMed  Google Scholar 

  4. Dardalhon V, Awasthi A, Kwon H, Galileos G, Gao W, Sobel RA, Mitsdoerffer M, Strom TB, Elyaman W, Ho IC, Khoury S, Oukka M, Kuchroo VK (2008) IL-4 inhibits TGF-beta-induced Foxp3+ T cells and, together with TGF-beta, generates IL-9+ IL-10+ Foxp3(−) effector T cells. Nat Immunol 9(12):1347–1355. doi:10.1038/ni.1677

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Chang HC, Sehra S, Goswami R, Yao W, Yu Q, Stritesky GL, Jabeen R, McKinley C, Ahyi AN, Han L, Nguyen ET, Robertson MJ, Perumal NB, Tepper RS, Nutt SL, Kaplan MH (2010) The transcription factor PU.1 is required for the development of IL-9-producing T cells and allergic inflammation. Nat Immunol 11(6):527–534. doi:10.1038/ni.1867

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Jabeen R, Kaplan MH (2012) The symphony of the ninth: the development and function of Th9 cells. Curr Opin Immunol 24(3):303–307. doi:10.1016/j.coi.2012.02.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Goswami R, Jabeen R, Yagi R, Pham D, Zhu J, Goenka S, Kaplan MH (2012) STAT6-dependent regulation of Th9 development. J Immunol 188(3):968–975. doi:10.4049/jimmunol.1102840

    Article  CAS  PubMed  Google Scholar 

  8. Staudt V, Bothur E, Klein M, Lingnau K, Reuter S, Grebe N, Gerlitzki B, Hoffmann M, Ulges A, Taube C, Dehzad N, Becker M, Stassen M, Steinborn A, Lohoff M, Schild H, Schmitt E, Bopp T (2010) Interferon-regulatory factor 4 is essential for the developmental program of T helper 9 cells. Immunity 33(2):192–202. doi:10.1016/j.immuni.2010.07.014

    Article  CAS  PubMed  Google Scholar 

  9. Jabeen R, Goswami R, Awe O, Kulkarni A, Nguyen ET, Attenasio A, Walsh D, Olson MR, Kim MH, Tepper RS, Sun J, Kim CH, Taparowsky EJ, Zhou B, Kaplan MH (2013) Th9 cell development requires a BATF-regulated transcriptional network. J Clin Invest 123(11):4641–4653. doi:10.1172/JCI69489

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Kaplan MH, Hufford MM, Olson MR (2015) The development and in vivo function of T helper 9 cells. Nat Rev Immunol 15(5):295–307. doi:10.1038/nri3824

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Lois C, Refaeli Y, Qin XF, Van Parijs L (2001) Retroviruses as tools to study the immune system. Curr Opin Immunol 13(4):496–504

    Article  CAS  PubMed  Google Scholar 

  12. Riviere I, Gallardo HF, Hagani AB, Sadelain M (2000) Retroviral-mediated gene transfer in primary murine and human T-lymphocytes. Mol Biotechnol 15(2):133–142. doi:10.1385/MB:15:2:133

    Article  CAS  PubMed  Google Scholar 

  13. Morita S, Kojima T, Kitamura T (2000) Plat-E: an efficient and stable system for transient packaging of retroviruses. Gene Ther 7(12):1063–1066. doi:10.1038/sj.gt.3301206

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. HB Wells Center for Pediatric Research, Indiana School of Medicine, 1044 W Walnut St., Indianapolis, IN, 46202, USA

    Rukhsana Jabeen Ph.D.

Authors
  1. Rukhsana Jabeen Ph.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rukhsana Jabeen Ph.D. .

Editor information

Editors and Affiliations

  1. School of Bio Science, Sir JC Bose Laboratory Complex, Indian Institute of Technology, Kharagpur, West Bengal, India

    Ritobrata Goswami

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Jabeen, R. (2017). Retroviral Transduction and Reporter Assay: Transcription Factor Cooperation in Th9 Cell Development. In: Goswami, R. (eds) Th9 Cells. Methods in Molecular Biology, vol 1585. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6877-0_12

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6877-0_12

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6876-3

  • Online ISBN: 978-1-4939-6877-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation