title: Fusion protein His-Hsp65-6IA2P2 prevents type 1 diabetes through nasal immunization in NOD Mice.
abstract: Human heat shock protein 60 (Hsp60), is an endogenous β-cells autoantigen, it could postpone the onset of insulitis and sooner type 1 diabetes mellitus. P277 is one of Hsp65 determinants at position 437-469 of amino acids cascaded. Meanwhile, it's already well-known that there were several better anti-diabetic B epitopes, such as insulinoma antigen-2 (IA-2). Currently, fusion protein IA2P2 has constructed in order to enhance its pharmacological efficacy. In addition, added homologous bacterial-derived Hsp65 and His tag were beneficial to protein immunogenicity and purification separately. So, finally we examined a fusion protein His-Hsp65-6IA2P2 could regulate Th2 immune response and reduce natural diabetic incidence in NOD mice. We constructed two express vector pET28a-His-Hsp65-6P277 and pET28a-His-Hsp65-6IA2P2. After purification, we observed that triple intranasal administration of these two fusion protein in 4-week-old NOD mice maintained normal blood glucose and weight, with a lower diabetic or insulitis incidence. Consistent with induced splenic T cells proliferation and tolerance, His-Hsp65-6IA2P2-treated mice performed reduced IFN-γ and increased IL-10 level. In conclusion, we suggested that fusion protein His-Hsp65-6IA2P2 could be reconstructed and purified successively. Furthermore, nasal administration of this fusion protein could rebalance T cells population and prevent T1DM.
authors.affiliation: Minigene Pharmacy Laboratory, School of Life Science & Technology, China Pharmaceutical University, PR China.
authors.forename: Shiping
authors.initials: S
authors.lastname: Lu
authors.affiliation: Minigene Pharmacy Laboratory, School of Life Science & Technology, China Pharmaceutical University, PR China.
authors.forename: Guoliang
authors.initials: G
authors.lastname: Li
authors.affiliation: Minigene Pharmacy Laboratory, School of Life Science & Technology, China Pharmaceutical University, PR China.
authors.forename: Kunfeng
authors.initials: K
authors.lastname: Liu
authors.affiliation: Minigene Pharmacy Laboratory, School of Life Science & Technology, China Pharmaceutical University, PR China.
authors.forename: Xue
authors.initials: X
authors.lastname: Yang
authors.affiliation: Minigene Pharmacy Laboratory, School of Life Science & Technology, China Pharmaceutical University, PR China.
authors.forename: Rongyue
authors.initials: R
authors.lastname: Cao
authors.affiliation: Institute of Pharmaceutics, School of Pharmacy, China Pharmaceutical University, PR China.
authors.forename: Li
authors.initials: L
authors.lastname: Zong
authors.affiliation: Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica,College of Pharmacy, Nanjing University of Chinese Medicine, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Long
authors.affiliation: Minigene Pharmacy Laboratory, School of Life Science & Technology, China Pharmaceutical University, PR China.
authors.forename: Liang
authors.initials: L
authors.lastname: Jin
authors.affiliation: Minigene Pharmacy Laboratory, School of Life Science & Technology, China Pharmaceutical University, PR China. Electronic address: wujie@cpu.edu.cn.
authors.forename: Jie
authors.initials: J
authors.lastname: Wu
mesh_terms: D000281:Administration, Intranasal; D000818:Animals; D002478:Cells, Cultured; D018834:Chaperonin 60 / Q000235:genetics; D016207:Cytokines / Q000378:metabolism; D003922:Diabetes Mellitus, Type 1 / Q000188:drug therapy*; D004195:Disease Models, Animal; D005260:Female; D006360:Heat-Shock Proteins / Q000235:genetics; D006639:Histidine / Q000235:genetics; D006801:Humans; D007114:Immunization; D051379:Mice; D016688:Mice, Inbred NOD; D010446:Peptide Fragments / Q000235:genetics; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8 / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000627:therapeutic use*; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D058408:Th1-Th2 Balance / Q000187:drug effects; D024101:Mitochondrial Proteins
chemical_list: D018834:Chaperonin 60; D016207:Cytokines; D006360:Heat-Shock Proteins; D006639:Histidine; D010446:Peptide Fragments; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8; D011993:Recombinant Fusion Proteins; C095402:peptide 277, heat shock protein 60; C516862:PTPRN protein, human; C060293:HSPD1 protein, human; D024101:Mitochondrial Proteins
keywords: Fusion protein; NOD mice; Nasal immunization; Type 1 diabetes mellitus