title: The Position of His-Tag in Recombinant OspC and Application of Various Adjuvants Affects the Intensity and Quality of Specific Antibody Response after Immunization of Experimental Mice.
abstract: Lyme disease, Borrelia burgdorferi-caused infection, if not recognized and appropriately treated by antibiotics, may lead to chronic complications, thus stressing the need for protective vaccine development. The immune protection is mediated by phagocytic cells and by Borrelia-specific complement-activating antibodies, associated with the Th1 immune response. Surface antigen OspC is involved in Borrelia spreading through the host body. Previously we reported that recombinant histidine tagged (His-tag) OspC (rOspC) could be attached onto liposome surfaces by metallochelation. Here we report that levels of OspC-specific antibodies vary substantially depending upon whether rOspC possesses an N' or C' terminal His-tag. This is the case in mice immunized: (a) with rOspC proteoliposomes containing adjuvants MPLA or non-pyrogenic MDP analogue MT06; (b) with free rOspC and Montanide PET GEL A; (c) with free rOspC and alum; or (d) with adjuvant-free rOspC. Stronger responses are noted with all N'-terminal His-tag rOspC formulations. OspC-specific Th1-type antibodies predominate post-immunization with rOspC proteoliposomes formulated with MPLA or MT06 adjuvants. Further analyses confirmed that the structural features of soluble N' and C' terminal His-tag rOspC and respective rOspC proteoliposomes are similar including their thermal stabilities at physiological temperatures. On the other hand, a change in the position of the rOspC His-tag from N' to C' terminal appears to affect substantially the immunogenicity of rOspC arguably due to steric hindrance of OspC epitopes by the C' terminal His-tag itself and not due to differences in overall conformations induced by changes in the His-tag position in rOspC variants.
authors: Department of Immunology, Faculty of Medicine and Dentistry, Palacky University Olomouc, Olomouc, Czech Republic.
authors: Michal
authors: M
authors: Krupka
authors: Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno, Czech Republic.
authors: Josef
authors: J
authors: Masek
authors: Department of Immunology, Faculty of Medicine and Dentistry, Palacky University Olomouc, Olomouc, Czech Republic.
authors: Lucia
authors: L
authors: Barkocziova
authors: Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno, Czech Republic.
authors: Pavlina
authors: P
authors: Turanek Knotigova
authors: Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno, Czech Republic.
authors: Pavel
authors: P
authors: Kulich
authors: Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno, Czech Republic.
authors: Jana
authors: J
authors: Plockova
authors: Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno, Czech Republic.
authors: Robert
authors: R
authors: Lukac
authors: Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno, Czech Republic.
authors: Eliska
authors: E
authors: Bartheldyova
authors: Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno, Czech Republic.|International Clinical Research Center, St. Anne´s University Hospital, Brno, Czech Republic.
authors: Stepan
authors: S
authors: Koudelka
authors: International Clinical Research Center, St. Anne´s University Hospital, Brno, Czech Republic.|Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment RECETOX, Masaryk University, Brno, Czech Republic.
authors: Radka
authors: R
authors: Chaloupkova
authors: Centre of the Region Hana for Biotechnological and Agricultural Research, Faculty of Science, Palacky University Olomouc, Olomouc, Czech Republic.
authors: Marek
authors: M
authors: Sebela
authors: Apigenex, Prague, Czech Republic.
authors: Daniel
authors: D
authors: Zyka
authors: Apigenex, Prague, Czech Republic.
authors: Ladislav
authors: L
authors: Droz
authors: Department of Chemistry of Natural Compounds University of Chemistry and Technology, Prague, Czech Republic.
authors: Roman
authors: R
authors: Effenberg
authors: Department of Chemistry of Natural Compounds University of Chemistry and Technology, Prague, Czech Republic.
authors: Miroslav
authors: M
authors: Ledvina
authors: King's College London, Institute of Pharmaceutical Science, London, United Kingdom, and GlobalAcorn Ltd, London, United Kingdom.
authors: Andrew D
authors: AD
authors: Miller
authors: Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno, Czech Republic.
authors: Jaroslav
authors: J
authors: Turanek
authors: Department of Immunology, Faculty of Medicine and Dentistry, Palacky University Olomouc, Olomouc, Czech Republic.|Department of Pharmacology and Immunotherapy, Veterinary Research Institute, Brno, Czech Republic.
authors: Milan
authors: M
authors: Raska
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000917:Antibody Formation / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D000942:Antigens, Bacterial / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D025065:Borrelia burgdorferi / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D007114:Immunization; D008193:Lyme Disease / Q000276:immunology; D051379:Mice; D023421:Models, Animal; D055550:Protein Stability; D017433:Protein Structure, Secondary; D011510:Proteolipids; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; C079616:OspC protein; D011510:Proteolipids; D011993:Recombinant Fusion Proteins; C020485:proteoliposomes