title: Production of canine soluble CD40 ligand to induce maturation of monocyte derived dendritic cells for cancer immunotherapy.
abstract: CD40 ligand (CD40L) expressed by activated T cells is shown to induce maturation of immature dendritic cells (DCs) and this maturation is a vital part in DC based tumor immunotherapy. We constructed an expression vector by cloning the extracellular domain of canine CD40L fused to the signal sequence of canine IL-12p40. When PBMCs were incubated with canine granulocyte-macrophage (GM) -CSF and IL-4, expression of CD86 was significantly elevated, but the majority of cells displayed the morphology of immature DCs. Following addition of the expressed canine soluble CD40L (csCD40L) to the DC-inducing culture, the cell morphology shifted to that of mature DCs, and expression of CD80, CD86, MHC class II and CD1a was significantly enhanced. This morphological change and enhancement of expression was observed even when the csCD40L was present only in the second half period of the culture. Furthermore, the csCD40L caused a significant increase in IL-12 production from DCs. These results show that the csCD40L significantly promotes the maturation and activation of canine monocyte derived DCs.
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano City, Osaka, Japan.
authors.forename: Viskam
authors.initials: V
authors.lastname: Wijewardana
authors.forename: Kikuya
authors.initials: K
authors.lastname: Sugiura
authors.forename: Mana
authors.initials: M
authors.lastname: Yahata
authors.forename: Takashi
authors.initials: T
authors.lastname: Akazawa
authors.forename: Daluthgamage Patsy H
authors.initials: DP
authors.lastname: Wijesekera
authors.forename: Shigeki
authors.initials: S
authors.lastname: Imamoto
authors.forename: Shingo
authors.initials: S
authors.lastname: Hatoya
authors.forename: Norimitsu
authors.initials: N
authors.lastname: Inoue
authors.forename: Toshio
authors.initials: T
authors.lastname: Inaba
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D023201:CD40 Ligand / Q000096:biosynthesis / Q000235:genetics*; D003713:Dendritic Cells / Q000276:immunology / Q000502:physiology*; D004285:Dogs; D007167:Immunotherapy; D053762:Interleukin-12 Subunit p40 / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D009000:Monocytes / Q000166:cytology*; D009369:Neoplasms / Q000628:therapy*; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*
chemical_list: D053762:Interleukin-12 Subunit p40; D011993:Recombinant Fusion Proteins; D023201:CD40 Ligand
keywords: Dendritic cells; Expression vector; Maturation; Soluble CD40 ligand