title: IL-2-independent generation of FOXP3(+)CD4(+)CD8(+)CD25(+) cytotoxic regulatory T cell lines from human umbilical cord blood.
abstract: OBJECTIVE
  Since the existence of mouse naturally occurring CD4(+)CD25(+) T regulatory (Treg) cells was demonstrated, a variety of human Treg subsets have been identified as distinct T cell populations. Here we show the establishment of novel Treg cell lines possessing unique characteristics.
  
  
  METHODS
  Novel Treg cell lines, designated HOZOT, were generated by coculturing human umbilical cord blood cells with mouse stromal cell lines in the absence of exogenous IL-2 or other cytokines. HOZOT were characterized and compared with CD4(+)CD25(+) Treg cells in terms of the CD phenotype, FOXP3 expression, suppressor activity against allogeneic MLR, anergy property, and IL-10 production.
  
  
  RESULTS
  HOZOT were generated and expanded as normal lymphoblastoid cells with cytotoxic activity against the cocultured stromal cells. HOZOT consisted of three subpopulations as defined by phenotype: CD4(+)CD8(+), CD4(+)CD8(dim), and CD4(-)CD8(+). All three subpopulations showed both suppressor and cytotoxic activities. While HOZOT's expression of FOXP3, CD25, GITR, and cytoplasmic CTLA-4 implied a similarity to naturally occurring CD4(+)CD25(+) Treg cells, these two Treg cells differed in IL-2 responsiveness and IL-10 production.
  
  
  CONCLUSIONS
  Our studies introduce a new method of generating Treg cells in an IL-2-independent manner and highlight a unique Treg cell type with cytotoxic activity and a phenotype of FOXP3(+)CD4(+)CD8(+)CD25(+).
authors.affiliation: Cell Biology Institute, Research Center, Hayashibara Biochemical Laboratories, Inc., Okayama, Okayama, Japan. shnakamu@hayashibara.co.jp
authors.forename: Shuji
authors.initials: S
authors.lastname: Nakamura
authors.forename: Motoyuki
authors.initials: M
authors.lastname: Suzuki
authors.forename: Akira
authors.initials: A
authors.lastname: Sugimoto
authors.forename: Kazue
authors.initials: K
authors.lastname: Tsuji-Takayama
authors.forename: Mayuko
authors.initials: M
authors.lastname: Yamamoto
authors.forename: Takeshi
authors.initials: T
authors.lastname: Otani
authors.forename: Toshiya
authors.initials: T
authors.lastname: Inoue
authors.forename: Akira
authors.initials: A
authors.lastname: Harashima
authors.forename: Ayumi
authors.initials: A
authors.lastname: Okochi
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Motoda
authors.forename: Fumiyuki
authors.initials: F
authors.lastname: Yamasaki
authors.forename: Kunzo
authors.initials: K
authors.lastname: Orita
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Kibata
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis*; D000943:Antigens, Differentiation / Q000096:biosynthesis; D060908:CTLA-4 Antigen; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques; D003601:Cytotoxicity Tests, Immunologic; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D051858:Forkhead Transcription Factors / Q000096:biosynthesis*; D053450:Glucocorticoid-Induced TNFR-Related Protein; D006801:Humans; D016130:Immunophenotyping; D016753:Interleukin-10 / Q000096:biosynthesis / Q000378:metabolism; D007376:Interleukin-2 / Q000494:pharmacology*; D051379:Mice; D017475:Receptors, Nerve Growth Factor / Q000096:biosynthesis; D018124:Receptors, Tumor Necrosis Factor / Q000096:biosynthesis; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D000943:Antigens, Differentiation; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; C556707:Ctla4 protein, mouse; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D053450:Glucocorticoid-Induced TNFR-Related Protein; C508609:IL10 protein, human; D007376:Interleukin-2; D017475:Receptors, Nerve Growth Factor; D018124:Receptors, Tumor Necrosis Factor; C491405:TNFRSF18 protein, human; D016753:Interleukin-10