Affinity-purified dengue-2 virus envelope glycoprotein induces neutralizing antibodies and protective immunity in mice.
We constructed a recombinant baculovirus which produces a dengue (DEN)-2 virus envelope (E) protein containing a six-histidine (H6) tag in place of the last 100 amino acids at its C-terminus. The recombinant protein was purified from the supernatant of baculovirus-infected Spodoptera frugiperda insect cell cultures to apparent homogeneity by cation-chelation chromatography (TALON) in which the H6-tagged E-protein was eluted under non-denaturing conditions with 100 mM imidazole at pH 8.0. Mice vaccinated with the purified E mixed with aluminium hydroxide adjuvant showed an immune response of IgM and IgG1, IgG2a and IgG2b isotypes, and neutralizing antibodies, similar to that following immunization with purified inactivated DEN-2 virus. Moreover, mice that received the purified recombinant protein were significantly protected against 200 50% lethal dose of DEN-2 virus. This combination of affinity purified H6-tagged protein and adsorption onto a cationic carrier seems promising for the production of immunogenic particulate proteins, especially DEN protein for the four serotypes, and the development of a new generation of vaccines.
Staropoli
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I
Unité des Arbovirus et Virus des Fièvres Hémorragiques, Institut Pasteur, Paris, France.
Frenkiel
M P
MP
Mégret
F
F
Deubel
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V
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis*; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D019281:Dimerization; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification / Q000627:therapeutic use; D019595:Severe Dengue / Q000276:immunology / Q000517:prevention & control*; D018411:Spodoptera / Q000821:virology; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000276:immunology* / Q000302:isolation & purification / Q000627:therapeutic use*
D000914:Antibodies, Viral; C051821:E-glycoprotein, Dengue virus type 2; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins