Autologous extracellular traps (ETs) influence T cell profiles and are not activating factors of human M1 macrophages
The formation of extracellular traps (ETs) is an important microbicidal functional mechanism of immune cells due to its implication in the pathogenesis of various diseases, including COVID-19. We set out to study the influence of ETs on the differentiation profiles of human lymphocytes and macrophag...
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| Autores principales: | , , , , , , |
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| Formato: | Artículo revista |
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Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
2022
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| Acceso en línea: | https://revistas.unc.edu.ar/index.php/med/article/view/39024 |
| Aporte de: |
| Sumario: | The formation of extracellular traps (ETs) is an important microbicidal functional mechanism of immune cells due to its implication in the pathogenesis of various diseases, including COVID-19. We set out to study the influence of ETs on the differentiation profiles of human lymphocytes and macrophages in autologous culture. The objectives were a) to cause the generation of ETs in vitro with lipopolysaccharide (LPS) or formylated peptides fMLFN-formyl-met-leu-phe (fMLP) and their isolation; b) to cause processing and presentation of the heterologous ovalbumin antigen (OVA) and c) mark by immunofluorescence (IF) molecules of the CD4 T profile, Th17 helper T cells and innate lymphoid cells 3 ILC3 (transcription factor RORɣ, orphan nuclear receptor); as well as the common leukocyte antigen CD45RO+ marker for T-cell activation and, the macrophage classic activation profile molecule M1, inducible nitric oxide synthetase (iNOS).
Autologous total leukocyte cell cultures were performed (n=10) from healthy human peripheral blood (donated by IHH, UNC, Ethically approved, HNC, FCM, UNC) that were challenged with OVA, and subjected to autologous ET stock. Controls: paired samples without challenge. Samples were taken at 24 and 72 h. IF with anti-RORɣ, anti-CD4, anti-CD45RO, anti-NOS2, and DNA staining with DAPI. Quantification of labeled cells was performed with FIJI program. Statistical treatment: t-test for paired samples.
At 24 hours, significant differences were observed in the percentages of positive cells for CD4 and CD45RO between paired control and OVA-challenged samples (*p<0.05) and between paired control and OVA- and ET-added samples (*p <0.05). In an independent experiment, significant differences were observed between those stimulated with OVA vs those stimulated with OVA and addition of ETs (*p<0.05). At 72 hours of culture, no significant differences were found between the paired samples in any case. There were no significant differences either at 24 or 72 hours of culture in the percentages of cells positive for RORɣ or in the percentages of cells positive for iNOS in different challenges.
Influence of ETs on T cell activation was observed and components of autologous ETs did not elicit classical activation of M1 macrophages. |
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