Use of Human Macrophages to Study Bunyavirus NSs Functions.
Methods Mol Biol
; 2824: 397-408, 2024.
Article
in En
| MEDLINE
| ID: mdl-39039426
ABSTRACT
The NSs protein is a major virulence factor in bunyaviruses, crucial for viral pathogenesis. However, assessing NSs protein function can be challenging due to its inhibition of cellular RNA polymerase II, impacting NSs protein expression from plasmid DNA. The recombinant Rift Valley fever virus (RVFV) MP-12 strain (rMP-12), a highly attenuated vaccine strain, can be safely manipulated under biosafety level 2 conditions. Leveraging a reverse genetics system, we can engineer rMP-12 variants expressing heterologous NSs genes, enabling functional testing in cultured cells. Human macrophages hold a central role in viral pathogenesis, making them an ideal model for assessing NSs protein functions. Consequently, we can comprehensively compare and analyze the functional significance of various NSs proteins in human macrophages using rMP-12 NSs variants. In this chapter, we provide a detailed overview of the preparation process for rMP-12 NSs variants and introduce two distinct human macrophage models THP-1 cells and primary macrophages. This research framework promises valuable insights into the virulence mechanisms of RVFV and other bunyaviruses and the potential for vaccine development.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Rift Valley fever virus
/
Viral Nonstructural Proteins
/
Macrophages
Limits:
Humans
Language:
En
Journal:
Methods Mol Biol
/
Methods in molecular biology
/
Methods mol. biol
Journal subject:
BIOLOGIA MOLECULAR
Year:
2024
Document type:
Article
Affiliation country:
United States
Country of publication:
United States