Preparation of hepatitis C virus structural and non-structural protein fragments and studies of their immunogenicity.
Plasmids pQE-60 and pQE-30 containing 6 x His-tag sequence were used for expression of fragments of HCV structural and non-structural proteins in Escherichia coli (E. coli). The following fragments were used: core (1-98 aa), NS3 (202-482 aa), and tetramer of hypervariable region 1 (HVR1) of E2 protein. The constructed plasmids directed high levels of expression of HCV proteins in E. coli JM109. After purification by the metal-affinity chromatography on nickel-nitrilotriacetic acid (Ni-NTA) agarose, the His-tagged HCV proteins were used for immunization of BALB/c mice. All three proteins were able to induce high levels of specific antibodies and, in the case of the NS3 and HVR1 tetramer, also to mount vigorous cell-proliferating responses. High immunogenicity of the tested fragments of HCV proteins shows them as good candidates for inclusion into the future HCV vaccine preparations.
Mihailova
Marija
M
Institute of Virology, Essen University Hospital, Hufelandstr. 55, 45122 Essen, Germany. mary@biomed.lu.lv
Fiedler
Melanie
M
Boos
Mechthild
M
Petrovskis
Ivars
I
Sominskaya
Irina
I
Roggendorf
Michael
M
Viazov
Sergei
S
Pumpens
Paul
P
D000818:Animals; D000937:Antigen-Antibody Reactions; D049109:Cell Proliferation; D002846:Chromatography, Affinity / Q000379:methods; D003001:Cloning, Molecular; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D018937:Hepatitis C Antibodies / Q000276:immunology; D018936:Hepatitis C Antigens / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D009532:Nickel / Q000737:chemistry; D009571:Nitrilotriacetic Acid / Q000737:chemistry; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D010455:Peptides / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013154:Spleen / Q000166:cytology / Q000276:immunology; D014758:Viral Core Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D009532:Nickel; D009571:Nitrilotriacetic Acid; D010446:Peptide Fragments; D010455:Peptides; D011994:Recombinant Proteins; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; C400562:HVR1 protein, Hepatitis C virus; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C516958:core protein (1-98), hepatitis C virus; C499352:core protein p22, Hepatitis C virus