A simple, robust flow cytometry-based whole blood assay for investigating sex differential interferon alpha production by plasmacytoid dendritic cells

J Immunol Methods. 2022 May:504:113263. doi: 10.1016/j.jim.2022.113263. Epub 2022 Mar 24.

Abstract

Central to sex differences observed in outcome from infection and vaccination is the innate immune response, and specifically production of type I interferons by plasmacytoid dendtiric cells (pDCs), the main producers of IFN-α. Evaluation of IFN-α production by pDCs is therefore critical for studies of innate immune function. However, reliable measurement of pDC IFN-α is hampered by reduced cell yields and cytokine production after cryopreservation or after even short delays in stimulating freshly isolated cells. We here describe a simple yet robust method for measuring IFN-α production in pDCs that preserves cell activation and cytokine production through immediate stimulation of whole blood and subsequent maintenance at 37 °C.

Keywords: Immune sex differences; Innate immunity; Interferon alpha; Plasmacytoid dendritic cell; Toll-like receptor 7.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Dendritic Cells*
  • Female
  • Flow Cytometry
  • Humans
  • Immunity, Innate
  • Interferon Type I*
  • Interferon-alpha
  • Male

Substances

  • Interferon Type I
  • Interferon-alpha