A dengue-2 Envelope fragment inserted within the structure of the P64k meningococcal protein carrier enables a functional immune response against the virus in mice.
A gene fragment encoding for the amino acids (aa) 286-426 from the dengue Envelope (E) protein was expressed in Escherichia coli as two forms of fusion proteins. In one case, the E fragment was fused to the first 45 aa of the P64k protein from Neisseria meningitidis (PD2) while, in the other, it was inserted within the lipoil-binding domain of the aforementioned bacterial protein (PD3). PD2 was obtained as insoluble form within the cytoplasm of the bacteria while PD3 was distributed equally as soluble and insoluble forms. The insoluble forms of each protein as well as the soluble fraction of PD3 were semipurified to test the antigenicity and the immunogenicity in mice. The forms containing the entire P64k protein exhibited the highest recognition with different polyclonal and monoclonal antibodies. Consequently, the neutralizing antibodies elicited by the recombinant proteins were higher in the case of PD3 forms than with PD2, independently of the solubility status. In addition, mice inoculated with the semipurified insoluble form of PD3 were partially protected against lethal challenge with dengue-2 virus, administered by intracerebral inoculation. The results suggested the folding and carrier capacity of the P64k protein over the E fragment, converting PD3 as an attractive vaccine candidate against dengue-2 virus.
Hermida
Lisset
L
División de Vacunas, Centro de Ingeniería Genética y Biotecnología, Apdo 6162, Habana 10600, Cuba. lisset.hermida@cigb.edu.cu
Rodríguez
Rayner
R
Lazo
Laura
L
Silva
Ricardo
R
Zulueta
Aída
A
Chinea
Glay
G
López
Carlos
C
Guzmán
María G
MG
Guillén
Gerardo
G
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D003715:Dengue / Q000517:prevention & control* / Q000821:virology; D003716:Dengue Virus / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
D000914:Antibodies, Viral; D001425:Bacterial Outer Membrane Proteins; C051821:E-glycoprotein, Dengue virus type 2; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C112623:lpdA protein, Neisseria meningitidis