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...
Guardado en:
| Autores principales: | , , , , , , |
|---|---|
| Formato: | Artículo revista |
| Lenguaje: | Español |
| Publicado: |
Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
2019
|
| Materias: | |
| Acceso en línea: | https://revistas.unc.edu.ar/index.php/med/article/view/25794 |
| Aporte de: |
| id |
I10-R327-article-25794 |
|---|---|
| record_format |
ojs |
| institution |
Universidad Nacional de Córdoba |
| institution_str |
I-10 |
| repository_str |
R-327 |
| container_title_str |
Revista de la Facultad de Ciencias Médicas de Córdoba |
| language |
Español |
| format |
Artículo revista |
| topic |
extracellular traps human leukocytes beta tubulin HLA-DR trampas extracelulares leucocitos humanos beta tubulina HLA-DR |
| spellingShingle |
extracellular traps human leukocytes beta tubulin HLA-DR trampas extracelulares leucocitos humanos beta tubulina HLA-DR Rinero, R Rodriguez, FM Carabajal-Miotti, CL Carabajal-Miotti, CL Ruiz de Frattari, S Vargas, AH Gonzalez-Silva, NE Novak, ITC HLA-DR and beta-tubulin in extracellular traps in autologous human leukocyte cultures stimulated in vitro |
| topic_facet |
extracellular traps human leukocytes beta tubulin HLA-DR trampas extracelulares leucocitos humanos beta tubulina HLA-DR |
| author |
Rinero, R Rodriguez, FM Carabajal-Miotti, CL Carabajal-Miotti, CL Ruiz de Frattari, S Vargas, AH Gonzalez-Silva, NE Novak, ITC |
| author_facet |
Rinero, R Rodriguez, FM Carabajal-Miotti, CL Carabajal-Miotti, CL Ruiz de Frattari, S Vargas, AH Gonzalez-Silva, NE Novak, ITC |
| author_sort |
Rinero, R |
| title |
HLA-DR and beta-tubulin in extracellular traps in autologous human leukocyte cultures stimulated in vitro |
| title_short |
HLA-DR and beta-tubulin in extracellular traps in autologous human leukocyte cultures stimulated in vitro |
| title_full |
HLA-DR and beta-tubulin in extracellular traps in autologous human leukocyte cultures stimulated in vitro |
| title_fullStr |
HLA-DR and beta-tubulin in extracellular traps in autologous human leukocyte cultures stimulated in vitro |
| title_full_unstemmed |
HLA-DR and beta-tubulin in extracellular traps in autologous human leukocyte cultures stimulated in vitro |
| title_sort |
hla-dr and beta-tubulin in extracellular traps in autologous human leukocyte cultures stimulated in vitro |
| description |
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. |
| publisher |
Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología |
| publishDate |
2019 |
| url |
https://revistas.unc.edu.ar/index.php/med/article/view/25794 |
| work_keys_str_mv |
AT rineror hladrandbetatubulininextracellulartrapsinautologoushumanleukocyteculturesstimulatedinvitro AT rodriguezfm hladrandbetatubulininextracellulartrapsinautologoushumanleukocyteculturesstimulatedinvitro AT carabajalmiotticl hladrandbetatubulininextracellulartrapsinautologoushumanleukocyteculturesstimulatedinvitro AT carabajalmiotticl hladrandbetatubulininextracellulartrapsinautologoushumanleukocyteculturesstimulatedinvitro AT ruizdefrattaris hladrandbetatubulininextracellulartrapsinautologoushumanleukocyteculturesstimulatedinvitro AT vargasah hladrandbetatubulininextracellulartrapsinautologoushumanleukocyteculturesstimulatedinvitro AT gonzalezsilvane hladrandbetatubulininextracellulartrapsinautologoushumanleukocyteculturesstimulatedinvitro AT novakitc hladrandbetatubulininextracellulartrapsinautologoushumanleukocyteculturesstimulatedinvitro AT rineror hladrybetatubulinaentrampasextracelularesencultivosdeleucocitosautologoshumanosestimuladosinvitro AT rodriguezfm hladrybetatubulinaentrampasextracelularesencultivosdeleucocitosautologoshumanosestimuladosinvitro AT carabajalmiotticl hladrybetatubulinaentrampasextracelularesencultivosdeleucocitosautologoshumanosestimuladosinvitro AT carabajalmiotticl hladrybetatubulinaentrampasextracelularesencultivosdeleucocitosautologoshumanosestimuladosinvitro AT ruizdefrattaris hladrybetatubulinaentrampasextracelularesencultivosdeleucocitosautologoshumanosestimuladosinvitro AT vargasah hladrybetatubulinaentrampasextracelularesencultivosdeleucocitosautologoshumanosestimuladosinvitro AT gonzalezsilvane hladrybetatubulinaentrampasextracelularesencultivosdeleucocitosautologoshumanosestimuladosinvitro AT novakitc hladrybetatubulinaentrampasextracelularesencultivosdeleucocitosautologoshumanosestimuladosinvitro |
| first_indexed |
2024-09-03T21:01:02Z |
| last_indexed |
2024-09-03T21:01:02Z |
| _version_ |
1809210160422322176 |
| spelling |
I10-R327-article-257942024-08-27T18:26:16Z HLA-DR and beta-tubulin in extracellular traps in autologous human leukocyte cultures stimulated in vitro HLA-DR y beta-tubulina en trampas extracelulares en cultivos de leucocitos autólogos humanos estimulados in vitro Rinero, R Rodriguez, FM Carabajal-Miotti, CL Carabajal-Miotti, CL Ruiz de Frattari, S Vargas, AH Gonzalez-Silva, NE Novak, ITC extracellular traps human leukocytes beta tubulin HLA-DR trampas extracelulares leucocitos humanos beta tubulina HLA-DR 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. Las trampas extracelulares (TEs) son estructuras de cromatina y proteínas intracelulares que se extruyen en leucocitos en condiciones inflamatorias. Fueron descriptas primeramente en neutrófilos por Brinkmann y col (2004) denominándolas “trampas extracelulares neutrófilas” (NETs). Las NETs además de su rol en mecanismos de defensa frente a microorganismos patógenos han sido implicadas en daño tisular, trombosis, autoinmunidad e incluso en inmunoedición de cáncer. Actualmente se conoce que otros leucocitos además de los neutrófilos son capaces de generar TEs. No todas las TEs son iguales, esto depende de la fuente de estimulación. Moléculas requeridas para la presentación de antígenos y activación de células T como HLA-DR (CMH clase II) pueden expresarse en neutrófilos estimulados. La presencia de beta-tubulina y HLA-DR no se ha informado en las TEs. El objetivo del presente trabajo fue generar TEs en cultivos de leucocitos expuestos a estimuladores como lipopolisacarido (LPS) o a péptidos formilados (fMLP) y llevar a cabo el marcado de beta-tubulina y HLA-DR. Cultivos autólogos de leucocitos de muestras de sangre humana sana (n = 10) con consentimiento informado (Comité de Ética del HNC, FCM), anticoagulados con heparina, fueron estimulados con 25 ng/ml de LPS o 0,25 ng/ml de fMLP, 30 minutos. Técnica de inmunofluorescencia con anticuerpos anti-beta tubulina y anti HLA-DR, tinción de ADN con DAPI. Muestras de sangre apareadas proporcionaron los controles. Las TEs fueron visualizadas por microscopia de Inmunofluorescencia y el porcentaje de células que liberan TEs fue calculado como promedio de cinco campos (400x) normalizado por número total de células. La estimulación con LPS o con fMLP incrementó el nivel basal de las TEs de leucocitos derivadas de donantes sanos (p<0,001, test t de Student para muestras apareadas). Moléculas beta-tubulina y HLA-DR fueron localizadas en las TEs de todos los grupos. La liberación de HLA-DR en las TEs puede influir en el entorno que favorece a la vía clase II de presentación de antígenos. La expresión de beta-tubulina y HLA-DR contribuye a una mejor comprensión de la composición de las TEs generadas por diferentes estimuladores. Esto puede tener importancia como objetivo terapéutico en enfermedades en cuya patogenia se encuentran implicadas las TEs. Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2019-10-17 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://revistas.unc.edu.ar/index.php/med/article/view/25794 Revista de la Facultad de Ciencias Médicas de Córdoba.; 2019: Suplemento JIC XX Revista de la Facultad de Ciencias Médicas de Córdoba; 2019: Suplemento JIC XX Revista da Faculdade de Ciências Médicas de Córdoba; 2019: Suplemento JIC XX 1853-0605 0014-6722 10.31053/1853.0605.v76.nSuplemento spa https://revistas.unc.edu.ar/index.php/med/article/view/25794/27562 Derechos de autor 2019 Universidad Nacional de Córdoba https://creativecommons.org/licenses/by-nc/4.0 |