Ex vivo mass cytometry analysis reveals a profound myeloid proinflammatory signature in psoriatic arthritis synovial fluid
| Autor: | Nicole Yager, Ash Maroof, Hussein Al-Mossawi, Paul Bowness, Julian C. Knight, Alicia Lledo Lara, Suzanne Cole, F Penkava |
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| Rok vydání: | 2021 |
| Předmět: |
Adult
Male 0301 basic medicine Chemokine Myeloid T-Lymphocytes medicine.medical_treatment T cell Immunology Enzyme-Linked Immunosorbent Assay CCL2 Monocytes General Biochemistry Genetics and Molecular Biology Proinflammatory cytokine 03 medical and health sciences 0302 clinical medicine Rheumatology Synovial Fluid medicine Humans Immunology and Allergy Synovial fluid Myeloid Cells RNA-Seq Osteopontin Chemokine CCL2 030203 arthritis & rheumatology biology business.industry Chemistry Gene Expression Profiling Macrophages Arthritis Psoriatic Dendritic Cells Dendritic cell Middle Aged Flow Cytometry Molecular biology 030104 developmental biology medicine.anatomical_structure Cytokine biology.protein Female Single-Cell Analysis business |
| DOI: | 10.1101/2021.03.04.433903 |
| Popis: | ObjectivesA number of immune populations have been implicated in psoriatic arthritis (PsA) pathogenesis. This study used mass cytometry (CyTOF) combined with transcriptomic analysis to generate a high-dimensional dataset of matched PsA synovial fluid (SF) and blood leucocytes, with the aim of identifying cytokine production ex vivo in unstimulated lymphoid and myeloid cells.MethodsFresh SF and paired blood were either fixed or incubated with protein transport inhibitors for 6 h. Samples were stained with two CyTOF panels: a phenotyping panel and an intracellular panel, including antibodies to both T cell and myeloid cell secreted proteins. Transcriptomic analysis by gene array of key expanded cell populations and single-cell RNA-seq, and ELISA and LEGENDplex analysis of PsA SF were also performed.ResultsWe observed marked changes in the myeloid compartment of PsA SF relative to blood, with expansion of intermediate monocytes, macrophages and dendritic cell populations. Classical monocytes, intermediate monocytes and macrophages spontaneously produced significant levels of the proinflammatory mediators osteopontin and CCL2 in the absence of any in vitro stimulation. By contrast minimal spontaneous cytokine production by T cells was detected. Gene expression analysis showed the genes for osteopontin and CCL2 to be amongst those most highly upregulated by PsA monocytes/macrophages; and both proteins were elevated in PsA SF.ConclusionsUsing multiomic analyses we have generated a comprehensive cellular map of PsA SF and blood to reveal key expanded myeloid proinflammatory modules in PsA of potential pathogenic and therapeutic importance. |
| Databáze: | OpenAIRE |
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