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

• Published in: PloS one Vol. 11; no. 2; p. e0148497
• Main Authors: Krupka, Michal, Masek, Josef, Barkocziova, Lucia, Turanek Knotigova, Pavlina, Kulich, Pavel, Plockova, Jana, Lukac, Robert, Bartheldyova, Eliska, Koudelka, Stepan, Chaloupkova, Radka, Sebela, Marek, Zyka, Daniel, Droz, Ladislav, Effenberg, Roman, Ledvina, Miroslav, Miller, Andrew D, Turanek, Jaroslav, Raska, Milan
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 05.02.2016; Public Library of Science (PLoS)
• Subjects: Adjuvants; Adjuvants, Immunologic; Alum; Aluminum sulfate; Amino acids; Animals; Antibiotics; Antibodies; Antibodies, Bacterial - immunology; Antibody Formation - immunology; Antibody response; Antibody Specificity - immunology; Antigens; Antigens, Bacterial - administration & dosage; Antigens, Bacterial - chemistry; Antigens, Bacterial - immunology; Bacterial Outer Membrane Proteins - administration & dosage; Bacterial Outer Membrane Proteins - chemistry; Bacterial Outer Membrane Proteins - immunology; Biology and Life Sciences; Borrelia; Borrelia burgdorferi; Borrelia burgdorferi - immunology; Causes of; Complications; Complications and side effects; Dentistry; Development and progression; Enzyme-Linked Immunosorbent Assay; Epitopes; Formulations; Genetic aspects; Histidine; Immune response; Immune system; Immunization; Immunogenicity; Immunoglobulins; Immunology; Immunotherapy; Lipids; Lyme disease; Lyme Disease - immunology; Lymphocytes T; Medicine and Health Sciences; Mice; Models, Animal; Phagocytes; Pharmacology; Physiological aspects; Prevention; Protein Stability; Protein Structure, Secondary; Proteins; Proteolipids; Recombinant Fusion Proteins - administration & dosage; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - isolation & purification; Research and Analysis Methods; Steric hindrance; Vaccine development; Vaccines; Vector-borne diseases; United Kingdom; Czech Republic
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0148497

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.