title: Identification and functional characterization of multiple interleukin 12 in amberjack (Seriola dumerili).
abstract: Interleukin (IL) -12 is a heterodimeric cytokine mainly produced by monocytes, macrophages, and dendritic cells in mammals. IL-12p70 composed of IL-12p35 and IL-12p40, is known to play a crucial role in promoting cell-mediated immunity (CMI) through Th1 differentiation and IFN-γ production. Although two types of IL-12p35 (p35a, p35b) and three types of IL-12p40 (p40a, p40b and p40c) have been identified in several fish species, the knowledge on functional characteristics of teleost IL-12 is still limited. In the present study, we cloned two types of IL-12p35 and three types of IL-12p40 genes in amberjack and yellowtail, and analyzed their expressions in response to stimulation with Nocardia seriolae in amberjack. As a result, four types of IL-12 (IL-12p35a, p35b, p40a and p40b) and IFN-γ mRNA were increased by live-N. seriolae stimulation but not by formalin-killed N. seriolae, suggesting that four types of IL-12 (p35, p35b, p40a and p40c) participate in promoting CMI. Subsequently, we produced six types of recombinant IL-12p70 (rIL12p70) protein in insect cells. Head kidney leukocytes were cultured with formalin-killed N. seriolae and six types of rIL-12p70 to elucidate the role of amberjack IL-12p70 in induction of CMI. After stimulation, IFN-γ expression was elevated whereas IL-10 expression was suppressed in Head kidney leukocytes stimulated with four types of rIL-12 (p40a/p35a, p40c/p35a, p40a/p35b, p40a/p35b). On the other hand, two types of rIL-12 (p40b/p35a, p40b/p35b) only elicited down regulation of IL-10 expression. These results indicate that all amberjack IL-12p70 isoforms are involved in Th1 -differentiation and promotion of CMI with different manners. Fish IL-12 has a potential for the promising vaccine adjuvant.
authors.affiliation: The United Graduate School of Agricultural Sciences, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-8580, Japan.
authors.forename: Megumi
authors.initials: M
authors.lastname: Matsumoto
authors.affiliation: Faculty of Fisheries, Kagoshima University, 4-50-20 Shimoarata, Kagoshima 890-0056, Japan.
authors.forename: Kazuma
authors.initials: K
authors.lastname: Hayashi
authors.affiliation: Department of Marine Bioscience, Fukui Prefectural University, Obama, Fukui 917-0003, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Suetake
authors.affiliation: Faculty of Fisheries, Kagoshima University, 4-50-20 Shimoarata, Kagoshima 890-0056, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Yamamoto
authors.affiliation: Faculty of Fisheries, Kagoshima University, 4-50-20 Shimoarata, Kagoshima 890-0056, Japan. Electronic address: araki@fish.kagoshima-u.ac.jp.
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Araki
mesh_terms: D000193:Actinomycetales Infections / Q000382:microbiology / Q000628:therapy / Q000662:veterinary*; D000595:Amino Acid Sequence; D000818:Animals; D001428:Bacterial Vaccines / Q000276:immunology*; D005393:Fish Diseases / Q000382:microbiology / Q000628:therapy*; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018664:Interleukin-12 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D053765:Interleukin-12 Subunit p35 / Q000235:genetics / Q000378:metabolism; D053762:Interleukin-12 Subunit p40 / Q000235:genetics / Q000378:metabolism; D009618:Nocardiaceae / Q000276:immunology*; D010473:Perciformes; D010802:Phylogeny; D016415:Sequence Alignment / Q000662:veterinary; D014613:Vaccines, Attenuated / Q000276:immunology; D015164:Vaccines, Inactivated / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology
chemical_list: D001428:Bacterial Vaccines; D029941:Fish Proteins; D053765:Interleukin-12 Subunit p35; D053762:Interleukin-12 Subunit p40; D014613:Vaccines, Attenuated; D015164:Vaccines, Inactivated; D014614:Vaccines, Synthetic; D018664:Interleukin-12
keywords: Amberjack; Cell-mediated immunity; IFNgamma; IL-12; Yellowtail