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

Oliver Sampson; Nicholas Lim; Jemima White; Vinicius Vieira; Henrik Kløverpris; Emily Adland; Chris Conlon; Donal Skelly; Lucy Jones; Lizzie Stafford; Anni Jamsen; Ellie Barnes; Susie Dunachie; John Frater; Paul Klenerman; Marcus Altfeld; Philip Goulder

doi:10.1016/j.jim.2022.113263

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.

Authors

Oliver Sampson¹, Nicholas Lim², Jemima White³, Vinicius Vieira⁴, Henrik Kløverpris⁵, Emily Adland⁶, Chris Conlon⁷, Donal Skelly⁸, Lucy Jones⁹, Lizzie Stafford¹⁰, Anni Jamsen¹¹, Ellie Barnes¹², Susie Dunachie¹³, John Frater¹⁴, Paul Klenerman¹⁵, Marcus Altfeld¹⁶, Philip Goulder¹⁷

Affiliations

¹ Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK. Electronic address: [email protected].
² Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK. Electronic address: [email protected].
³ Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK. Electronic address: [email protected].
⁴ Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK. Electronic address: [email protected].
⁵ Africa Health Research Institute, Nelson R. Mandela School of Medicine, K-RITH Tower Building, Umbilo Road, Durban, South Africa. Electronic address: [email protected].
⁶ Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK. Electronic address: [email protected].
⁷ Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, OX3 9DU, England, UK. Electronic address: [email protected].
⁸ Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK; Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, OX3 9DU, England, UK; Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, Oxford OX3 9DU, England, UK. Electronic address: [email protected].
⁹ Department of Integrated Sexual Health, Cwm Taf Morgannwg University Health Board, Pontypridd, CF37 1LB, Wales, UK. Electronic address: [email protected].
¹⁰ Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, OX3 9DU, England, UK. Electronic address: [email protected].
¹¹ Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, OX3 9DU, England, UK. Electronic address: [email protected].
¹² Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK; Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, OX3 9DU, England, UK. Electronic address: [email protected].
¹³ Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK; Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, OX3 9DU, England, UK. Electronic address: [email protected].
¹⁴ Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK; Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, OX3 9DU, England, UK. Electronic address: [email protected].
¹⁵ Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK; Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, OX3 9DU, England, UK. Electronic address: [email protected].
¹⁶ Research Department Virus Immunology, Leibniz Institute for Experimental Virology, Hamburg, Germany. Electronic address: [email protected].
¹⁷ Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, England, UK. Electronic address: [email protected].

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

Abstract

Publication types

MeSH terms

Substances

Grants and funding