Establishment of a neutralization assay for Nipah virus using a high-titer pseudovirus system.
Biotechnol Lett
; 45(4): 489-498, 2023 Apr.
Article
en En
| MEDLINE
| ID: mdl-36680637
ABSTRACT
OBJECTIVE:
To construct a high-titer Nipah pseudovirus packaging system using the HIV lentivirus backbone vector and establish a safe neutralization assay for Nipah pseudovirus in biosafety level 2 facilities.METHODS:
Nipah virus (NiV) fusion protein (F) and glycoprotein (G) recombinant expression plasmids, psPAX2, and pLenti CMV Puro LUC (w168-1) were transiently transfected into 293T cells for 72 h for the generation of a NiV pseudovirus. The neutralization ability of Nipah virus F and G protein antibodies was assessed using the pseudovirus.RESULTS:
A NiV pseudovirus was constructed using 293T cells. The ideal mass ratio of plasmid psPAX2 w168-1 F G for transfection was determined to be 4411. The specificity of recombinant F and G protein expression was indicated by indirect immunofluorescence and western blotting. The pseudovirus particles showed obvious spikes under a transmission electron microscope. The NiV pseudovirus titer was 4.73 × 105 median tissue culture infective dose per mL, and the pseudovirus could be effectively neutralized by polyclonal antibodies specifically targeting the F and G proteins respectively.CONCLUSIONS:
A NiV pseudovirus was successfully generated using HIV vector systems, and was used as a platform for a safe and reliable pseudovirus-based neutralizing assay that can be performed in biosafety level 2 facilities.Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Infecciones por VIH
/
Virus Nipah
Límite:
Humans
Idioma:
En
Revista:
Biotechnol Lett
Año:
2023
Tipo del documento:
Article
País de afiliación:
China