Chapter title |
Polarizing Cytokines for Human Th9 Cell Differentiation
|
---|---|
Chapter number | 6 |
Book title |
Th9 Cells
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6877-0_6 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6876-3, 978-1-4939-6877-0
|
Authors |
Prabhakar Putheti M.S., Ph.D., Prabhakar Putheti |
Editors |
Ritobrata Goswami |
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. |
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