Abstract
CD4+ T helper (Th) cell subset generation in vivo requires T cell receptor activation and surface CD28 co-stimulation in the presence of one or more cytokines. Similarly, Th cells can be generated in vitro by activating naïve CD4+CD25− T cells with plate bound-anti-CD3 monoclonal antibody (mAb) (pbCD3) and soluble-anti-CD28 mAb (sCD28) in the presence of polarizing recombinant (r) cytokines and anti-cytokine mAbs. In comparison to in vitro CD4+CD25− T cells, memory CD4+CD25−CD45RO+ T cells have been shown to convert to Th9 cells more efficiently. Here, protocol for in vitro generation of human Th9 cells by activating CD4+CD25−CD45RO+ memory T cells with pbCD3 and sCD28 in the presence of polarizing recombinant interleukin-4 (rIL-4) and transforming growth factor (rTGF-β) is described.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Tada T, Takemori T, Okumura K, Nonaka M, Tokuhisa T (1978) Two distinct types of helper T cells involved in the secondary antibody response: independent and synergistic effects of Ia− and Ia+ helper T cells. J Exp Med 147(2):446–458
Mosmann TR, Cherwinski H, Bond MW, Giedlin MA, Coffman RL (1986) Two types of murine helper T cell clone I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol 136(7):2348–2357
Szabo SJ, Sullivan BM, Peng SL, Glimcher LH (2003) Molecular mechanisms regulating Th1 immune responses. Annu Rev Immunol 21:713–758
Ansel KM, Djuretic I, Tanasa B, Rao A (2006) Regulation of Th2 differentiation and Il4 l0Cus accessibility. Annu Rev Immunol 24:607–656
Harrington LE, Hatton RD, Mangan PR, Turner H, Murphy TL, Murphy KM, Weaver CT (2005) Interleukin 17-producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages. Nat Immunol 6(11):1123–1132
Park H, Li Z, Yang XO, Chang SH, Nurieva R, Wang YH, Wang Y, Hood L, Zhu Z, Tian Q, Dong C (2005) A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nat Immunol 6(11):1133–1141
Weaver CT, Hatton RD, Mangan PR, Harrington LE (2007) IL-17 family cytokines and the expanding diversity of effector T cell lineages. Annu Rev Immunol 25:821–852
Sakaguchi S (2004) Naturally arising CD4+ regulatory t cells for immunologic self-tolerance and negative control of immune responses. Annu Rev Immunol 22:531–562
Chen W, Jin W, Hardegen N, Lei KJ, Li L, Marinos N, McGrady G, Wahl SM (2003) Conversion of peripheral CD4+CD25− naive T cells to CD4+CD25+ regulatory T cells by TGF-beta induction of transcription factor Foxp3. J Exp Med 198(12):1875–1886
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
Chang HC, Han L, Jabeen R, Carotta S, Nutt SL, Kaplan MH (2009) PU.1 regulates TCR expression by modulating GATA-3 activity. J Immunol 183(8):4887–4894
Putheti P, Awasthi A, Popoola J, Gao W, Strom TB (2010) Human CD4 memory T cells can become CD4+IL-9+ T cells. PLoS One 5(1):e8706
Dardalhon V, Korn T, Kuchroo VK, Anderson AC (2008) Role of Th1 and Th17 cells in organ-specific autoimmunity. J Autoimmun 31(3):252–256
Putheti P, Pettersson A, Soderstrom M, Link H, Huang YM (2004) Circulating CD4+CD25+ T regulatory cells are not altered in multiple sclerosis and unaffected by disease-modulating drugs. J Clin Immunol 24(2):155–161
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Putheti, P. (2017). Polarizing Cytokines for Human Th9 Cell Differentiation. 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_6
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6877-0_6
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