title: Synthetic peptides containing B- and T-cell epitope of dengue virus-2 E domain III provoked B- and T-cell responses.
abstract: Our previous work applied a combination of bioinformatics approaches and in vitro assays to identify the dengue-2 virus (DENV-2)-specific B- and T-cell epitopes. In this report, we first evaluated the antigenicity of both B- and T-cell epitopes reacting with different sera against DENV-2 by ELISA as well as the ability of T-cell epitope to activate CD4(+) T-cell producing IFN-γ using ELISPOT, which showed a specific reactivity between either B- or T-cell epitope and DENV-2 antisera, and a significant increase of IFN-γ producing cells in DENV-2 infected mice. Then, a multi-epitope peptide containing the above B-, T-cell epitopes of envelope domain III (EDIII) of DENV-2 and pan-DR epitope (PADRE) was bioinformatically designed and synthesized. The verification of its immunogenicity and protective effect was performed in in vitro and in vivo experiments. The results showed that a high level of antibody in mice elicited by the multi-epitope peptide was detected by ELISA and the anti-peptide sera binding to the vero cells infected with DEN-2 was observed with immunofluorescence test. More importantly, the peptide could induce lymphoproliferation in vitro and a predominant Th1 type of immune response was examined by flow cytometry. We also found that the virus replication in the mice vaccinated with the multi-epitope peptide was obviously less than that of the control groups. These results may provide some important information for the development of dengue vaccine.
authors.affiliation: Department of Microbiology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China.
authors.forename: Shanfeng
authors.initials: S
authors.lastname: Li
authors.forename: Liang
authors.initials: L
authors.lastname: Peng
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
authors.forename: Hua
authors.initials: H
authors.lastname: Zhong
authors.forename: Fuchun
authors.initials: F
authors.lastname: Zhang
authors.forename: Ziqiang
authors.initials: Z
authors.lastname: Yan
authors.forename: Hong
authors.initials: H
authors.lastname: Cao
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D002522:Chlorocebus aethiops; D003715:Dengue / Q000276:immunology / Q000517:prevention & control; D053059:Dengue Vaccines / Q000276:immunology*; D003716:Dengue Virus / Q000276:immunology*; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005260:Female; D007111:Immunity, Cellular; D056724:Immunity, Humoral; D007114:Immunization; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D007371:Interferon-gamma / Q000097:blood; D051379:Mice; D008810:Mice, Inbred C57BL; D009500:Neutralization Tests; D012367:RNA, Viral / Q000097:blood; D022223:Vaccines, Subunit / Q000235:genetics / Q000276:immunology; D014709:Vero Cells
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D053059:Dengue Vaccines; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D007074:Immunoglobulin G; D012367:RNA, Viral; D022223:Vaccines, Subunit; D007371:Interferon-gamma