A novel fusion protein domain III-capsid from dengue-2, in a highly aggregated form, induces a functional immune response and protection in mice.
Based on the immunogenicity of domain III from the Envelope protein of dengue virus as well as the proven protective capacity of the capsid antigen, we have designed a novel domain III-capsid chimeric protein with the goal of obtaining a molecule potentially able to induce both humoral and cell-mediated immunity (CMI). After expression of the recombinant gene in Escherichia coli, the domain III moiety retained its antigenicity as evaluated with anti-dengue sera. In order to explore alternatives for modulating the immunogenicity of the protein, it was mixed with oligodeoxynucleotides in order to obtain particulated aggregates and then immunologically evaluated in mice in comparison with non-aggregated controls. Although the humoral immune response induced by both forms of the protein was equivalent, the aggregated variant resulted in a much stronger CMI as measured by in vitro IFN-gamma secretion and protection experiments, mediated by CD4(+) and CD8(+) cells. The present work provides additional evidence in support for a crucial role of CMI in protection against dengue virus and describes a novel vaccine candidate against the disease based on a recombinant protein that can stimulate both arms of the acquired immune system.
Valdés
Iris
I
Vaccine Division, Center for Genetic Engineering and Biotechnology (CIGB), Ave 31, P.O. Box 6162, Havana 6, 10 600, Cuba. iris.valdes@cigb.edu.cu
Bernardo
Lidice
L
Gil
Lázaro
L
Pavón
Alekis
A
Lazo
Laura
L
López
Carlos
C
Romero
Yaremis
Y
Menendez
Ivón
I
Falcón
Viviana
V
Betancourt
Lázaro
L
Martín
Jorge
J
Chinea
Glay
G
Silva
Ricardo
R
Guzmán
María G
MG
Guillén
Gerardo
G
Hermida
Lisset
L
D000818:Animals; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D003715:Dengue / Q000276:immunology* / Q000517:prevention & control*; D003716:Dengue Virus / Q000145:classification / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity; D005260:Female; D007371:Interferon-gamma / Q000096:biosynthesis; D015847:Interleukin-4 / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D046529:Microscopy, Electron, Transmission; D046912:Multiprotein Complexes; D010957:Plasmids / Q000235:genetics; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000956:Antigens, Viral; D036022:Capsid Proteins; D046912:Multiprotein Complexes; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins; D015847:Interleukin-4; D007371:Interferon-gamma