title: DENV-2 subunit proteins fused to CR2 receptor-binding domain (P28)-induces specific and neutralizing antibodies to the Dengue virus in mice.
abstract: Domain III (DIII) of the dengue virus (DENV) envelope (E) protein induces strong neutralizing type-specific antibodies. In addition, a region near the fusion loop in domain II (DII) induces the production of cross-reactive antibodies with neutralizing potential. Thus, this study aimed to generate DENV-2 recombinant fusion proteins (i.e., rEII*EIII and rEII*EIII/NS1*) either alone or fused to 3 copies of P28, the minimum CR2-binding domain of the complement protein C3d. The 4 recombinant proteins were generated in a Drosophila melanogaster Schneider 2 (S2) cell system. The expression and secretion of the recombinant proteins were confirmed in vitro using immunofluorescence (IF) and western blot (WB) analyses. Human dengue immune serum samples recognized recombinant proteins. The immunogenicity of the 4 proteins in BALB/c mice was analyzed using ELISA, and the results revealed that the induced specific antibody response was higher in the groups of mice immunized with the P28 fusion proteins. Interestingly, although the 4 recombinant proteins were able to elicit high levels of neutralizing antibodies in BALB/c mice; no adjuvant effect was observed in terms of neutralizing antibodies in the groups immunized with proteins containing P28. Thus, ELISA and PRNT50 assays may evaluate different epitopes and responses, where ELISA showed a wider response that did not always correlate with neutralization. Furthermore, the elicited antibodies were able to recognize the immobilized E glycoprotein of DENV. All mice vaccinated with the DENV-2 recombinant proteins showed induction of higher levels of IgG1 antibodies than of IgG2a antibodies.
authors.affiliation: Department of Molecular Biomedicine Centre for Research and Advanced Studies (CINVESTAV-IPN) Av. IPN # 2508 Col.; San Pedro Zacatenco, D.F. Mexico, Mexico.
authors.forename: Jazmín
authors.initials: J
authors.lastname: García-Machorro
authors.forename: Moisés
authors.initials: M
authors.lastname: López-González
authors.forename: Olivia
authors.initials: O
authors.lastname: Barrios-Rojas
authors.forename: Cynthia
authors.initials: C
authors.lastname: Fernández-Pomares
authors.forename: Claudia
authors.initials: C
authors.lastname: Sandoval-Montes
authors.forename: Leopoldo
authors.initials: L
authors.lastname: Santos-Argumedo
authors.forename: Nicolás
authors.initials: N
authors.lastname: Villegas-Sepúlveda
authors.forename: Benito
authors.initials: B
authors.lastname: Gutiérrez-Castañeda
authors.forename: Leticia
authors.initials: L
authors.lastname: Cedillo-Barrón
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood*; D002460:Cell Line; D015933:Complement C3d / Q000235:genetics / Q000378:metabolism*; D053059:Dengue Vaccines / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology*; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004331:Drosophila melanogaster; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D011485:Protein Binding; D011951:Receptors, Complement / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D053059:Dengue Vaccines; D011951:Receptors, Complement; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D015933:Complement C3d
keywords: dengue enveloped protein Domain II and III; immunization; recombinant proteins