Characterization of secreted and intracellular forms of a truncated hepatitis C virus E2 protein expressed by a recombinant herpes simplex virus.
A replication-defective herpes simplex virus type 1 (HSV-1) recombinant lacking the glycoprotein H (gH)-encoding gene and expressing a truncated form of the hepatitis C (HCV) E2 glycoprotein (E2-661) was constructed and characterized. We show here that cells infected with the HSV/HCV recombinant virus efficiently express the HCV E2-661 protein. Most importantly, cellular and secreted E2-661 protein were both readily detected by the E2-conformational mAb H53 and despite the high expression levels, only limited amounts of misfolded aggregates were detected in either the cellular or secreted fractions. Furthermore, cell-associated and secreted E2-661 protein bound to the major extracellular loop (MEL) of CD81 in a concentration-dependent manner and both were highly reactive with sera from HCV-infected patients. Finally, BALB/c mice immunized intraperitoneally with the recombinant HSV/HCV virus induced high levels of anti-E2 antibodies. Analysis of the induced immunoglobulin G (IgG) isotypes showed high levels of IgG2a while the levels of the IgG1 isotype were significantly lower, suggesting a Th1-type of response. We conclude that the HSV-1 recombinant virus represents a promising tool for production of non-aggregated, immunologically active forms of the E2-661 protein and might have potential applications in vaccine development.
Lucas
M
M
Molecular Virology Laboratory, Hellenic Pasteur Institute, 127 Vas. Sofias Ave, Athens 115 21, Greece.
Tsitoura
E
E
Molecular Virology Laboratory, Hellenic Pasteur Institute, 127 Vas. Sofias Ave, Athens 115 21, Greece.
Montoya
M
M
The Edward Jenner Institute for Vaccine Research, Compton, UK.
Laliotou
B
B
Molecular Virology Laboratory, Hellenic Pasteur Institute, 127 Vas. Sofias Ave, Athens 115 21, Greece.
Aslanoglou
E
E
Molecular Virology Laboratory, Hellenic Pasteur Institute, 127 Vas. Sofias Ave, Athens 115 21, Greece.
Kouvatsis
V
V
Molecular Virology Laboratory, Hellenic Pasteur Institute, 127 Vas. Sofias Ave, Athens 115 21, Greece.
Entwisle
C
C
Xenova Group plc, Berkshire, UK.
Miller
J
J
Xenova Group plc, Berkshire, UK.
Klenerman
P
P
Nuffield Department of Medicine, University of Oxford, Oxford, UK.
Hadziyannis
A
A
Second Department of Medicine, Athens University School of Medicine, Greece.
Hadziyannis
S
S
Second Department of Medicine, Athens University School of Medicine, Greece.
Borrow
P
P
The Edward Jenner Institute for Vaccine Research, Compton, UK.
Mavromara
P
P
Molecular Virology Laboratory, Hellenic Pasteur Institute, 127 Vas. Sofias Ave, Athens 115 21, Greece.
D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D002460:Cell Line; D005260:Female; D018937:Hepatitis C Antibodies / Q000032:analysis* / Q000097:blood; D018936:Hepatitis C Antigens / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D019698:Hepatitis C, Chronic / Q000097:blood / Q000517:prevention & control; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis; D018139:Simplexvirus / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014611:Vaccination; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics* / Q000276:immunology*
D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C044200:glycoprotein H, herpes simplex virus type 1; C091015:glycoprotein E2, Hepatitis C virus