[Antiviral activity determination of recombinant equine interferon-gamma and identification inhibited antiviral activity of monoclonal antibodies].
Equine interferon-gamma (eIFN-gamma) expressed both in E. coli and baculovirus were evaluated for antiviral activity against recombinant Vesicular Stomatits Virus expressing green fluorescence protein (rVSV-GFP) in EFK-78 cells. The assays were conducted in 96-well plate. Virus infectivity was measured by quantifying GFP-positive cells, instead of quantifying the CPE reduction. Prior to infection of EFK-78 cells with rVSV-GFP, the cells were incubated with eIFN-gamma. The GFP expression in the EFK-78 cells dramatically decreased in the cells treated with eIFN-gamma in a dose-dependent manner, comparing with the mock-treated cells. The titers of antiviral activity were 1 x 10(3) AU/mL and 1 x 10(5) AU/mL of eIFN-gamma expressed from E. coli and baculovirus, respectively. The antiviral activities of the recombinant eIFN-gamma were highly efficient and specific, as it was blocked by mAbs against eIFN-gamma.
Bai
Yu
Y
National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin 150001, China.
Chen
Weiye
W
Tong
Tiegang
T
Zhang
Weijun
W
Xu
Shulan
S
Wang
Qun
Q
Sun
Qingge
Q
Liu
Guangliang
G
Bu
Zhigao
Z
Wu
Donglai
D
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology*; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006736:Horses; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology*; D011994:Recombinant Proteins; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma