title: Rapid ex vivo isolation and long-term culture of human Th17 cells.
abstract: 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 RORgammat 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 RORgammat, 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.
authors.affiliation: Partners AIDS Research Center, Division of Infectious Diseases, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02129, United States. hstreeck@partners.org
authors.forename: Hendrik
authors.initials: H
authors.lastname: Streeck
authors.forename: Kristin W
authors.initials: KW
authors.lastname: Cohen
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Jolin
authors.forename: Mark A
authors.initials: MA
authors.lastname: Brockman
authors.forename: Angela
authors.initials: A
authors.lastname: Meier
authors.forename: Karen A
authors.initials: KA
authors.lastname: Power
authors.forename: Michael T
authors.initials: MT
authors.lastname: Waring
authors.forename: Galit
authors.initials: G
authors.lastname: Alter
authors.forename: Marcus
authors.initials: M
authors.lastname: Altfeld
mesh_terms: D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D020869:Gene Expression Profiling; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods; D020381:Interleukin-17 / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D054398:Receptors, CCR4 / Q000096:biosynthesis / Q000235:genetics; D018123:Receptors, Interleukin / Q000096:biosynthesis / Q000235:genetics; D018168:Receptors, Retinoic Acid / Q000096:biosynthesis / Q000235:genetics; D011988:Receptors, Thyroid Hormone / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology* / Q000276:immunology
chemical_list: D000906:Antibodies; C515354:CCR4 protein, human; C515500:IL23R protein, human; D020381:Interleukin-17; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D012333:RNA, Messenger; C539900:RORC protein, human; D054398:Receptors, CCR4; D018123:Receptors, Interleukin; D018168:Receptors, Retinoic Acid; D011988:Receptors, Thyroid Hormone