title: Identification of a cell line producing high levels of TSLP: advantages for screening of anti-allergic drugs.
abstract: Thymic stromal lymphopoietin (TSLP) plays critical roles in the induction and exacerbation of allergic diseases. These findings suggest that an inhibitor of TSLP production may be a novel drug for allergic diseases. However, conducting high-throughput screening of such compounds is difficult because there is currently no appropriate in vitro system. In the present study, we demonstrated that the mouse keratinocyte cell line KCMH-1 produced higher amounts of TSLP than the mouse keratinocyte cell line PAM-212, human keratinocyte cell line HaCaT, or bronchial cell line BEAS-2B. A reporter gene assay revealed that transcriptional activity of the TSLP gene was also markedly higher in KCMH-1 than in PAM212 cells. Both dexamethasone and the retinoid X receptor agonist HX600 inhibited the production of TSLP in KCMH-1 cells, which indicated that its production could be pharmacologically regulated. Moreover, the biological activity of TSLP released from KCMH-1 cells in the medium was endorsed by the induction of OX40L expression in bone marrow-derived dendritic cells. These results indicate that KCMH-1 can be utilized in high-throughput screening of inhibitors of TSLP production and also as a source of native TSLP.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Segawa
authors.affiliation: Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
authors.forename: Saori
authors.initials: S
authors.lastname: Yamashita
authors.affiliation: Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
authors.forename: Natsumi
authors.initials: N
authors.lastname: Mizuno
authors.affiliation: Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
authors.forename: Mika
authors.initials: M
authors.lastname: Shiraki
authors.affiliation: Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Hatayama
authors.affiliation: Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
authors.forename: Nozomi
authors.initials: N
authors.lastname: Satou
authors.affiliation: Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Hiratsuka
authors.affiliation: Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima 734-8551, Japan.
authors.forename: Michihiro
authors.initials: M
authors.lastname: Hide
authors.affiliation: Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan. Electronic address: hirasawa@m.tohoku.ac.jp.
authors.forename: Noriyasu
authors.initials: N
authors.lastname: Hirasawa
mesh_terms: D000818:Animals; D018926:Anti-Allergic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D003713:Dendritic Cells / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology; D003984:Dibenzazepines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D057166:High-Throughput Screening Assays; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008808:Mice, Inbred CBA; D041681:NIH 3T3 Cells; D053278:OX40 Ligand; D019899:Paracrine Communication; D012333:RNA, Messenger / Q000378:metabolism; D047488:Retinoid X Receptors / Q000819:agonists / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D048069:Tumor Necrosis Factors / Q000235:genetics / Q000378:metabolism; D000094632:Thymic Stromal Lymphopoietin
chemical_list: D018926:Anti-Allergic Agents; D016207:Cytokines; D003984:Dibenzazepines; D008562:Membrane Glycoproteins; D053278:OX40 Ligand; D012333:RNA, Messenger; D047488:Retinoid X Receptors; C504586:Tnfsf4 protein, mouse; D048069:Tumor Necrosis Factors; C104181:HX 600; D003907:Dexamethasone; D000094632:Thymic Stromal Lymphopoietin
keywords: Dexamethasone; Keratinocyte; OX40L; Thymic stromal lymphopoietin