Impact of epitope permutations in the antibody response of mice to a multi-epitope polypeptide of the V3 loop of human immunodeficiency virus type 1.
Our group have produced in Escherichia coli and evaluated the immunogenicity of different multi-epitope polypeptides (MEPs) bearing one copy of V3 loop sequential B cell epitopes from several isolates of human immunodeficiency virus type 1 (HIV-1) gp120. One of these MEPs called TAB9 comprises the 15 central amino acids of the V3 loop from isolates LR150, JY1, RF, MN, BRVA and IIIB in this order. Antibodies against all V3 regions were elicited after immunization of rabbits, macaques and humans with TAB9. In contrast, mice immunized with this protein only developed antibodies against epitopes JY1, LR150 and MN in that order (JY1>LR150>MN>>>RF, BRVA, IIIB) resembling an immunodominant gradient from the N-terminus to the C-terminal portion of this construction. To assess what role the location of the V3 epitopes in TAB9 could play, we constructed the protein TAB16, by altering the position of V3 epitopes in TAB9 primary structure and compared the pattern of antibodies elicited by both MEPs in H-2(d) Balb/c mice. The MEP TAB16 elicited antibody titers comparable to that of the sera from mice immunized with TAB9. There were no statistical differences in antibody titers between both groups (P>0.05). JY1, LR150 and MN V3 epitopes were again immunodominant in mice immunized with TAB16 fusion protein. The highest antibody titers detected in both groups among V3 epitopes corresponded to JY1, now located at the C-terminus of the permuted chimera. Antibodies against V3 epitopes RF, BRVA and IIIB were again not detected. Additionally, the MN V3 epitope showed to be significantly more immunogenic in its new orientation in TAB16, possibly as a result of a higher degree of accessibility in the surface of the protein. The results of the present investigation strongly suggest that the sequential order or the intramolecular position of V3 epitopes inside the primary structure of TAB9 and TAB16 MEPs does not interfere with the global immunogenicity or with the hierarchy of immunodominance of these regions.
Aguilar
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A
Departamento de SIDA, Centro de Ingeniería Genética y Biotecnología, Apdo 6162, Cubanacán, Playa, 10600, Havana, Cuba.
Carrazana
Y
Y
Duarte
C A
CA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D005260:Female; D015699:HIV Envelope Protein gp120 / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000737:chemistry* / Q000276:immunology; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D018985:Epitopes, B-Lymphocyte; D015699:HIV Envelope Protein gp120; C059762:HIV envelope protein gp120 (305-321); D016056:Immunodominant Epitopes; D007135:Immunoglobulin Variable Region; D010446:Peptide Fragments; D010455:Peptides; D011993:Recombinant Fusion Proteins; C417682:TAB9