SARS-CoV-2 Spike Protein Activates Human Lung Macrophages
Abstract
:1. Introduction
2. Results
2.1. Effects of Spike Protein on Cytokine and Chemokine Release from HLMs
2.2. Effects of Spike Protein on Ca2+ Homeostasis
2.3. Effects of Spike Protein on Morphology and Kinetic Properties of HLMs
2.4. Effects of Spike Protein on Phagocytosis
2.5. Expression of ACE2 and TMPRSS2 on HLMs
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Isolation and Purification of Human Lung Macrophages (HLMs)
4.3. SARS-CoV-2 Spike Glycoprotein Stimulation of HLMs
4.4. Cell Viability
4.5. ELISA Assays
4.6. RT-PCR
4.7. [Ca2+]i Measurement in Single-Cell
4.8. Time-Lapse and High-Content Microscopy
4.9. Phagocytosis Assay
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
ACE | angiotensin-converting enzyme |
ANGPT | angiopoietin |
Ct | cycle threshold |
Ca2+ | calcium concentrations |
FCS | fetal calf serum |
HLM | human lung macrophage |
IL- | interleukin |
LPS | lipopolysaccharide |
MDM | monocyte-derived macrophage |
PBS | phosphate buffer saline |
PIPES | piperazine- N, N_-bis-2-ethanesulphonic acid |
RT-qPCR | quantitative reverse transcriptase PCR |
RFU | relative fluorescent unit |
TNF | tumor necrosis factor |
VEGF | vascular endothelial growth factor |
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Palestra, F.; Poto, R.; Ciardi, R.; Opromolla, G.; Secondo, A.; Tedeschi, V.; Ferrara, A.L.; Di Crescenzo, R.M.; Galdiero, M.R.; Cristinziano, L.; et al. SARS-CoV-2 Spike Protein Activates Human Lung Macrophages. Int. J. Mol. Sci. 2023, 24, 3036. https://doi.org/10.3390/ijms24033036
Palestra F, Poto R, Ciardi R, Opromolla G, Secondo A, Tedeschi V, Ferrara AL, Di Crescenzo RM, Galdiero MR, Cristinziano L, et al. SARS-CoV-2 Spike Protein Activates Human Lung Macrophages. International Journal of Molecular Sciences. 2023; 24(3):3036. https://doi.org/10.3390/ijms24033036
Chicago/Turabian StylePalestra, Francesco, Remo Poto, Renato Ciardi, Giorgia Opromolla, Agnese Secondo, Valentina Tedeschi, Anne Lise Ferrara, Rosa Maria Di Crescenzo, Maria Rosaria Galdiero, Leonardo Cristinziano, and et al. 2023. "SARS-CoV-2 Spike Protein Activates Human Lung Macrophages" International Journal of Molecular Sciences 24, no. 3: 3036. https://doi.org/10.3390/ijms24033036