Immune response to recombinant visna virus Gag and Env precursor proteins synthesized in insect cells.
Two different recombinant visna virus (VV) gag-baculoviruses were constructed for the expression of precursor VV Gag in insect cells. Both recombinant Gag viruses expressed proteins migrating on SDS PAGE at the predicted rate for VV Gag precursor, Pr50gag. However, differences were seen in the morphology of the virus-like particles produced. Monoclonal antibody directed against the VV Gag capsid protein (p25) and sera from sheep infected with ovine lentiviruses reacted to both 50-kDa proteins. A recombinant VV env-baculovirus was constructed, substituting sequences encoding the signal peptide of VV Env with the murine IFN-gamma analogue. Sera from ovine lentivirus infected sheep reacted in immunoblots with two proteins of approximately 100 and 200 kDa found in the plasma membrane of insect cells infected with env-recombinant virus. Sheep immunized with either the recombinant Gag or the Env proteins developed high antibody titers to VV in ELISA. The serum of sheep and ascitic fluid of mice immunized with the recombinant Gag reacted with native Pr50gag and the processed Gag proteins in immunoblots, whereas serum of the recombinant Env immunized sheep reacted with VV gp135 and a putative oligomer of gp135. The immunized sheep responded specifically to visna virus by lymphocyte proliferation in vitro.
Rafnar
B
B
Institute for Experimental Pathology, University of Iceland, Reykjavik.
Tobin
G J
GJ
Nagashima
K
K
Gonda
M A
MA
Gunnarsson
E
E
Andrésson
O S
OS
Georgsson
G
G
Torsteinsdóttir
S
S
D000818:Animals; D002460:Cell Line; D015686:Gene Products, env / Q000235:genetics / Q000276:immunology*; D015683:Gene Products, gag / Q000235:genetics / Q000276:immunology*; D007114:Immunization; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D011498:Protein Precursors / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012756:Sheep; D018411:Spodoptera; D014790:Visna-maedi virus / Q000276:immunology*
D015686:Gene Products, env; D015683:Gene Products, gag; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins