Gene Expression
Nebulas
Gene Expression NebulasSummary: PTX3 is an essential component of humoral innate immunity, involved in resistance to selected pathogens and in the regulation of inflammation1-3. The present study was designed to assess the presence and significance of PTX3 in COVID-194-7. RNA-seq analysis of peripheral blood mononuclear cells, single-cell bioinformatics analysis and immunohistochemistry of lung autopsy samples revealed that myelomonocytic cells and endothelial cells express high levels of PTX3 in COVID-19 patients. Increased plasma concentrations of PTX3 were detected in 96 COVID-19 patients. PTX3 emerged as a strong independent predictor of 28-day mortality in multivariable analysis, better than conventional markers of inflammation, in hospitalized COVID-19 patients. The prognostic significance of PTX3 abundance for mortality was confirmed in a second independent cohort (54 patients). Thus, circulating and lung myelomonocytic cells and endothelial cells are a major source of PTX3, and PTX3 plasma concentration can serve as an independent strong prognostic indicator of short-term mortality in COVID-19.
Overall Design: Monocytes were isolated from peripheral blood samples of three healthy donors and six COVID-19 patients and RNA expression was measured with total RNA-Seq.
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| Growth Protocol: | - |
| Treatment Protocol: | - |
| Extract Protocol: | Monocytes were enriched from Ficoll-isolated PBMCs by positive selection using CD14 Microbeads kit (Miltenyi Biotec), RNA was purified with the Direct-zol RNA Microprep or Miniprep Kit (Zymosan) |
| Library Construction Protocol: | Total-RNA-sequencing library preparation was performed starting from 1 ng of total-RNA with the SMART-Seq Stranded Kit (Clontech-Takara). Libraries obtained were qualitatively assessed by using TapeStation 4200 (Agilent) and quantified by Qubit Fluorimeter (Thermo Fisher Scientific). Afterwards, they were multiplexed in equimolar pools and sequenced on a NextSeq-550 Illumina Platform generating at least 80 million 75-bp-PE reads per sample |
| Molecule Type: | rRNA- RNA |
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| Library Layout: | PAIRED |
| Library Strand: | Forward |
| Platform: | ILLUMINA |
| Instrument Model: | Illumina NextSeq 550 |
| Strand-Specific: | Specific |