HLA-DR and beta-tubulin in extracellular traps in autologous human leukocyte cultures stimulated in vitro
Extracellular traps (ETs) are chromatin structures and intracellular proteins that are extruded into leukocytes under inflammatory conditions. They were first described in neutrophils by Brinkmann et al (2004) calling them "neutrophil extracellular traps" (NETs). NETs, in addition to the...
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| Autores principales: | , , , , , , |
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| Formato: | Artículo revista |
| Lenguaje: | Español |
| Publicado: |
Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
2019
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| Materias: | |
| Acceso en línea: | https://revistas.unc.edu.ar/index.php/med/article/view/25794 |
| Aporte de: |
| Sumario: | Extracellular traps (ETs) are chromatin structures and intracellular proteins that are extruded into leukocytes under inflammatory conditions. They were first described in neutrophils by Brinkmann et al (2004) calling them "neutrophil extracellular traps" (NETs). NETs, in addition to their role in defense mechanisms against pathogenic microorganisms, have been implicated in tissue damage, thrombosis, autoimmunity and even in cancer immunoedition. It is currently known that other leukocytes besides neutrophils are capable of generating ETs. Not all ETs are equal, this depends on the source of stimulation. Molecules required for antigen presentation and activation of T cells such as HLA-DR (CMH class II) can be expressed in stimulated neutrophils. The presence of beta-tubulin and HLA-DR has not been reported in ETs. The aim of this work was to generate ETs in leukocyte cultures exposed to stimulants such as lipopolysaccharide (LPS) or to formylated peptides (fMLP) and carry out the labeling of beta-tubulin and HLA-DR.
Autologous leukocyte cultures of healthy human blood samples (n = 10) with informed consent (HNC Ethics Committee, FCM), anticoagulated with heparin, were stimulated with 25 ng / ml of LPS or 0.25 ng / ml of fMLP, 30 minutes. Immunofluorescence technique with anti-beta tubulin and anti HLA-DR antibodies, DNA staining with DAPI. Paired blood samples provided controls. The ETs were visualized by immunofluorescence microscopy and the percentage of cells that release ETs was calculated as an average of five fields (400x) normalized by total number of cells.
Stimulation with LPS or fMLP increased the baseline level of leukocyte ETs derived from healthy donors (p<0.001, Student's t-test for paired samples). Beta-tubulin and HLA-DR molecules were located in the ETs of all groups.
The release of HLA-DR in ETs can influence the environment that favors the class II pathway of antigen presentation. The expression of beta-tubulin and HLA-DR contributes to a better understanding of the composition of the ETs generated by different stimulators. This may be important as a therapeutic objective in diseases in which ETs are involved in their pathogenesis. |
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