title: Study on the immune response to recombinant Hsp70 protein from Megalobrama amblycephala.
abstract: The expression of heat shock protein 70 (Hsp70) is induced in response to many factors including high temperature, infection, metal pollutants and toxic chemicals. In this study, Megalobrama amblycephala HSP70 promoter was cloned, and characteristic heat shock elements (HSEs) were identified in the promoter region. The recombinant M. amblycephala Hsp70 protein (rMaHsp70) was expressed and purified from Escherichia coli BL21 (DE3). To evaluate in vivo immune response of rMaHsp70, we administered intraperitoneal (IP) injection, and demonstrated that rMaHsp70 stimulated M. amblycephala immune activity by inducing the expression of HSP70, HIF-1α, HSC70, CXCR4b, TNF-α and IL-1β mRNAs in liver, headkidney, spleen and gill, as well as SOD, glutathione, lysozyme and interferon alpha proteins in serum and liver. The effect of rMaHsp70 as adjuvant against Aeromonas hydrophila was assessed by injecting a mixed vaccine of rMaHsp70 and A. hydrophila (A. hydrophila/Hsp70) into M. amblycephala, and the relative percent survival (RPS) in the A. hydrophila/Hsp70 group was 75% compared to 50% in the A. hydrophila/PBS group. Furthermore, rMaHsp70 also promoted the proliferation and suppressed apoptosis in M. amblycephala fin cells (MAF) in a dose-dependent manner. Taken together, these results suggest that rMaHsp70 can induce organic immune response and improve environmental tolerance.
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070 Wuhan, PR China; Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, 430070 Wuhan, PR China.
authors.forename: Nan
authors.initials: N
authors.lastname: Chen
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070 Wuhan, PR China.
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Wan
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070 Wuhan, PR China; Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, 430070 Wuhan, PR China.
authors.forename: Chun-Xiao
authors.initials: CX
authors.lastname: Huang
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070 Wuhan, PR China; Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, 430070 Wuhan, PR China.
authors.forename: Wei-Min
authors.initials: WM
authors.lastname: Wang
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070 Wuhan, PR China; Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, 430070 Wuhan, PR China.
authors.forename: Hong
authors.initials: H
authors.lastname: Liu
authors.affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, 430070 Wuhan, PR China; Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, 430070 Wuhan, PR China. Electronic address: hbauwhl@hotmail.com.
authors.forename: Huan-Ling
authors.initials: HL
authors.lastname: Wang
mesh_terms: D024506:5' Flanking Region; D000276:Adjuvants, Immunologic; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D003530:Cyprinidae; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D059372:Nucleotide Motifs; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D017422:Sequence Analysis, DNA
chemical_list: D000276:Adjuvants, Immunologic; D018840:HSP70 Heat-Shock Proteins; D011994:Recombinant Proteins
keywords: Immune response; Megalobrama amblycephala; Recombinant Hsp70