title: The influence of concentration of inactivated Edwardsiella tarda bacterin and immersion time on antigen uptake and expression of immune-related genes in Japanese flounder (Paralichthys olivaceus).
abstract: Our previous work has demonstrated that the immune response of Japanese flounder was associated with the concentration of formalin-inactivated Edwardsiella tarda and immersion time. In order to further investigate the influence of immersion vaccine dose and bath time on the antigen uptake, formalin-killed Edwardsiella tarda bacterin was prepared and adjusted to four concentrations (109, 108, 107, 106cfu ml-1) for 30, 60 and 90 min immersion in Japanese flounder model, respectively. Absolute quantitative real-time PCR was employed to examine the bacterin uptake in gill, skin, spleen and kidney at 3 and 6 h post vaccination. The results showed that the antigen uptaken in gills and skin were significant higher than spleen and kidney, and the antigen amounts in gill and skin both declined from 3 to 6 h, whereas the antigen amounts in spleen and kidney gradually increased. Significant higher antigen amounts were detected in 109-30, 109-60, 108-60, 108-90 and 108-90 groups than other groups (P < 0.05), especially the 108-60min group displayed the highest antigen uptaken. Meanwhile, the expression profiles of antigen recognization and presentation genes (MHCⅡα, TcRα, CD4-1), immunoglobulins (IgM, IgT), inflammatory cytokines (IL-1β, IL-6), heat shock protein 70 (HSP70) and c-type lysozyme were analyzed using real-time PCR. On the whole, the transcription levels of the eight genes exhibited to be higher in 107-90, 108and 109cfu ml-1groups than other groups (P < 0.05), especially the 108-60 group displayed the highest up-regulation. These results demonstrated that immersion with formalin-inactivated E. tarda, especially under 108-60 min condition could efficiently enhance the antigen uptake and the expression of immune-related genes, which provided evidences for an enhanced vaccination effects under an optimized combination of vaccine dose and immersion time.
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Du
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.
authors.forename: Xiaoqian
authors.initials: X
authors.lastname: Tang
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.
authors.forename: Xiuzhen
authors.initials: X
authors.lastname: Sheng
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xing
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, 5 Yushan Road, Qingdao 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, No.1 Wenhai Road, Aoshanwei Town, Jimo, Qingdao 266071, China. Electronic address: wbzhan@ouc.edu.cn.
authors.forename: Wenbin
authors.initials: W
authors.lastname: Zhan
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial / Q000276:immunology; D001428:Bacterial Vaccines / Q000276:immunology*; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D020609:Edwardsiella tarda / Q000276:immunology*; D005393:Fish Diseases / Q000517:prevention & control*; D005432:Flounder / Q000235:genetics* / Q000276:immunology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D050884:HSP72 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D007114:Immunization; D007075:Immunoglobulin M / Q000235:genetics / Q000276:immunology; D018836:Inflammation Mediators / Q000378:metabolism; D009113:Muramidase / Q000235:genetics; D014613:Vaccines, Attenuated / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D016207:Cytokines; D050884:HSP72 Heat-Shock Proteins; D007075:Immunoglobulin M; D018836:Inflammation Mediators; D014613:Vaccines, Attenuated; D009113:Muramidase
keywords: Antigen uptake; Edwardsiella tarda; Gene expression; Immersion vaccination; Japanese flounder (Paralichthys olivaceus)