Use of Human Macrophages to Study Bunyavirus NSs Functions

Cigdem Alkan, Tetsuro Ikegami

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages397-408
Number of pages12
DOIs
StatePublished - 2024
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2824
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Bunyavirus
  • Human primary macrophages
  • NSs protein
  • Pathogenesis
  • Reverse genetics
  • THP-1 macrophages
  • Vaccines

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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