Rapid ex vivo isolation and long-term culture of human Th17 cells
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- 作者：Hendrik Streeck ; Kristin W. Cohen ; Jonathan S. Jolin ; Mark A. Brockman ; Angela Meier ; Karen A. Power ; Michael T. Waring ; Galit Alter ; Marcus Altfeld
- 关键词：Th17 cells ; Isolation ; Capture assay ; Long-term culture ; RORγ ; t
- 刊名：Journal of Immunological Methods
- 出版年：2008
- 出版时间：20 April 2008
- 年：2008
- 卷：333
- 期：1-2
- 页码：115-125
- 全文大小：1255 K
文摘
T helper (Th) 17 cells are a distinct lineage of CD4+ T cells mediating tissue inflammation through the secretion of IL-17. In addition, it has been shown that the expression of the transcriptional factor RORγt is responsible for the induction and maintenance of this cell line. Th17 cells are believed to be involved in a variety of autoimmune disorders, but may also play an important role in host defense. Here we describe a novel technique to reproducibly isolate viable Th17 cells based on their IL-17 secreting ability. We confirmed Th17 cell enrichment by quantitative PCR analysis and demonstrate that positively selected cells using this technique express significantly increased mRNA levels of RORγt, IL-23 receptor and CCR4 when compared to negatively selected cells. Furthermore, we show that purified Th17 cells can be maintained in long-term culture and expand in vitro. In conclusion, this technique will allow for the first time the direct, ex vivo analysis of phenotypic and functional properties of Th17 cells.