title: Generation of canine dendritic cells from peripheral blood monocytes without using purified cytokines.
abstract: Dendritic cells (DCs), which differentiate in vitro from peripheral blood monocytes (PBMOs) or bone marrow precursors, are a promising candidate for immunotherapy against cancer. The dog, which suffers common types of cancers along with humans, make an ideal large animal model for cancer studies. Monocyte-derived DCs in the dog have not been well characterized, however, since the appropriate condition for in vitro differentiation has not been established. To tackle this problem, we have developed a conditioned media by culturing T cells with immobilized anti-canine CD3 antibody, and sought to induce differentiation of DCs from PBMOs. When purified CD14+ PBMOs were cultured in the presence of 25% T cell conditioned medium (TCCM), the PBMOs increased size and had extended dendritic processes by day 12 of the culture. The cultured PBMOs were found to increase the expression of MHC class II and CD1a molecules, and significantly increased stimulatory activity for allogeneic T cells in the mixed leukocyte reaction. Moreover, the cells significantly increased their expression of IL-18 and IFN-gamma when stimulated with polyinosinic-polycytidylic acid (Poly (I:C)). The cells have a reduced phagocytic activity, which is a common defect in mature DCs. It follows from these results that TCCM does induce the differentiation of DCs from PBMOs.
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai City, Osaka 599-8531, Japan.
authors.forename: Viskam
authors.initials: V
authors.lastname: Wijewardana
authors.forename: Kikuya
authors.initials: K
authors.lastname: Sugiura
authors.forename: Tomoko
authors.initials: T
authors.lastname: Oichi
authors.forename: Mariko
authors.initials: M
authors.lastname: Fujimoto
authors.forename: Takashi
authors.initials: T
authors.lastname: Akazawa
authors.forename: Shingo
authors.initials: S
authors.lastname: Hatoya
authors.forename: Muneo
authors.initials: M
authors.lastname: Inaba
authors.forename: Susumu
authors.initials: S
authors.lastname: Ikehara
authors.forename: Thotawaththege S P
authors.initials: TS
authors.lastname: Jayaweera
authors.forename: Toshio
authors.initials: T
authors.lastname: Inaba
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods / Q000662:veterinary; D002454:Cell Differentiation / Q000276:immunology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016207:Cytokines / Q000235:genetics / Q000276:immunology; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D004285:Dogs / Q000097:blood / Q000276:immunology*; D004705:Endocytosis / Q000276:immunology; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology*; D007959:Lymphocyte Culture Test, Mixed / Q000662:veterinary; D018613:Microscopy, Confocal / Q000662:veterinary; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D000949:Histocompatibility Antigens Class II; D012313:RNA