title: Production of a recombinant cholera toxin B subunit-insulin B chain peptide hybrid protein by Brevibacillus choshinensis expression system as a nasal vaccine against autoimmune diabetes.
abstract: Mucosally induced tolerance is an attractive strategy for preventing or reducing autoimmune diseases. Here, we produced a recombinant CTB fusion protein linked with autoantigen T cell epitope of insulin B chain peptide 9-23 (C19S) at levels up to 200 mg/L culture media in Brevibacillus choshinensis secretion-expression system. Receptor-competitive assay showed that the CTB-insulin peptide binds to GM1 receptor almost equivalent degree as the native form of CTB. Non-obese diabetes (NOD) mice that spontaneously develop an insulin-dependent diabetes were nasally immunized with CTB-insulin peptide (5 microg) for three times. The nasal treatment significantly reduced the development of insulin-dependent diabetes and peptide specific DTH responses after systemic immunization with the insulin peptide B 9-23(C19S) in CFA. Nasal administration of as high as 50 microg of the peptide alone demonstrated a similar level of the disease inhibition. In contrast, all mice given 5 microg of the insulin peptide alone or 5 microg of insulin peptide with 25 microg of the free form of CTB did not lead to the suppression of diabetes development and DTH responses. Because molecular weight of the insulin peptide is about one tenth of that of the CTB-insulin peptide, the results demonstrate that the recombinant hybrid of autoantigen and CTB increased its tolerogenic potential for nasal administration by up 100-fold on molar base of autoantigen peptide. Taken together, nasally-induced tolerance by administration of the recombinant B. choshinensis-derived hybrid protein of CTB and autoantigen T cell-epitope peptide could be useful mucosal immunetherapy for the control of T cell-mediated autoimmune diseases.
authors.affiliation: Division of Mucosal Immunology, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 Japan. yukiy@ims.u-tokyo.ac.jp
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Yuki
authors.forename: Chisato
authors.initials: C
authors.lastname: Hara-Yakoyama
authors.forename: Abigail A E
authors.initials: AA
authors.lastname: Guadiz
authors.forename: Shigezo
authors.initials: S
authors.lastname: Udaka
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kiyono
authors.forename: Subhendra
authors.initials: S
authors.lastname: Chatterjee
mesh_terms: D000281:Administration, Intranasal; D000818:Animals; D001324:Autoantigens / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics; D001951:Brevibacterium; D002772:Cholera Toxin / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D003922:Diabetes Mellitus, Type 1 / Q000517:prevention & control; D018984:Epitopes, T-Lymphocyte / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D015870:Gene Expression; D007108:Immune Tolerance / Q000187:drug effects; D007328:Insulin / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D016688:Mice, Inbred NOD; D010455:Peptides / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D001324:Autoantigens; D018984:Epitopes, T-Lymphocyte; D007328:Insulin; D010455:Peptides; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; C034993:ganglioside receptor; D002772:Cholera Toxin