Efecto de Prevotella copri sobre la viabilidad, activación y diferenciación de monocitos y macrófagos
No hay miniatura disponible
Archivos
Fecha
2023
Autores
Profesor/a Guía
Facultad/escuela
Idioma
es
Título de la revista
ISSN de la revista
Título del volumen
Editor
Universidad Andrés Bello
Nombre de Curso
Licencia CC
Licencia CC
Resumen
La microbiota intestinal es una comunidad de diversos géneros
bacterianos comensales que interactúan con el individuo en el que habitan,
causando impacto en el sistema inmune. Sin embargo, su disbiosis se asocia
con el desarrollo de patologías inflamatorias como la Artritis Reumatoide (AR),
enfermedad autoinmune que causa la degradación de cartílago y material óseo
en las articulaciones sinoviales. Varios estudios han relacionado la AR a la
especie Prevotella copri (P. copri) encontrándose sobreabundante en la
microbiota intestinal de pacientes con AR temprana y pre-clínica. Se ha
demostrado que P. copri promueve el desarrollo de artritis experimental en
ratones SKG e induce un aumento de linfocitos Th17. Además, P. copri reduce
el grosor de la capa protectora de la mucosa intestinal. Esto sugiere la
participación de P. copri en el desarrollo y progresión de AR. Para dilucidar el
mecanismo por el cual P. copri podría contribuir a la inmunopatogénesis de la
AR, este proyecto se enfoca en investigar los efectos de P. copri sobre monocitos
y macrófagos humanos. Dado que se ha reportado que pacientes con AR activa
presentan un sesgo hacia macrófagos proinflamatorios M1, se hipotetiza que P.
copri reduce la viabilidad, promueve la activación y diferenciación de monocitos
y macrófagos hacia un perfil M1 in vitro. El objetivo general es evaluar el efecto
de P. copri sobre la viabilidad, activación y diferenciación de monocitos y
macrófagos in vitro. Como modelo de monocitos se empleó la línea celular THP1, que puede diferenciarse a macrófagos al ser expuesta a PMA. Se determinó
el efecto de P. copri sobre la viabilidad de monocitos y macrófagos in vitro,
mediante marcaje con Anexina V y una sonda de viabilidad mediante citometría
de flujo. Luego, se evaluó el efecto de P. copri sobre la activación y diferenciación
de monocitos y macrófagos in vitro, mediante el análisis de marcadores para
M1/M2 y del perfil de citoquinas por citometría de flujo. Este estudio contribuye
al conocimiento sobre la interacción entre P. copri y el sistema inmune innato,
particularmente los monocitos y macrófagos, en la inmunopatogénesis de AR,
que podrían asociarse al desarrollo y propagación de inflamación crónica en AR.
The intestinal microbiota is a community of diverse commensal bacterial genera that interact with the individual they inhabit, causing impact on the immune system. However, its dysbiosis is associated with the development of inflammatory pathologies, such as Rheumatoid Arthritis (RA), autoimmune disease that causes degradation of the cartilage and bone material in synovial joints. Several studies have associated RA with the specie Prevotella copri (P. copri), which is found overaccumulated in the intestinal microbiota of patients with early and pre-clinical RA. P. copri has been shown to promote the development of experimental arthritis in SKG mice and induce an increase in Th17 lymphocytes. In addition, P. copri is capable of reducing the thickness of the protective layer of the intestinal mucosa. These suggest a role of P. copri in the development and progression of RA. In order to elucidate the mechanism by which P. copri could contribute to the immunopathogenesis of RA, this project focuses on investigating the effects of P. copri on human monocytes and macrophages. Since it has been reported that patients with active RA have a bias towards proinflammatory M1 type macrophages, it is hypothesized that P. copri reduces viability and promotes the activation and differentiation of monocytes and macrophages towards an M1 profile in vitro. The general objective is to evaluate the effect of P. copri on the viability, activation and differentiation of monocytes and macrophages in vitro. The THP-1 cell line was used as a model for monocytes, which can differentiate into macrophages when exposed to PMA. The effect of P. copri on monocyte and macrophage viability in vitro by using Annexin V labeling and a viability probe by flow cytometry. Then, the effect of P. copri on the activation and differentiation of monocytes and macrophages in vitro was evaluated through the analysis of markers for M1/M2 and cytokine profiling by flow cytometry. This study contributes to the knowledge about the interaction between P. copri and the innate immune system, particularly monocytes and macrophages, in the immunopathogenesis of RA, which could be associated with the development and spread of chronic inflammation in RA.
The intestinal microbiota is a community of diverse commensal bacterial genera that interact with the individual they inhabit, causing impact on the immune system. However, its dysbiosis is associated with the development of inflammatory pathologies, such as Rheumatoid Arthritis (RA), autoimmune disease that causes degradation of the cartilage and bone material in synovial joints. Several studies have associated RA with the specie Prevotella copri (P. copri), which is found overaccumulated in the intestinal microbiota of patients with early and pre-clinical RA. P. copri has been shown to promote the development of experimental arthritis in SKG mice and induce an increase in Th17 lymphocytes. In addition, P. copri is capable of reducing the thickness of the protective layer of the intestinal mucosa. These suggest a role of P. copri in the development and progression of RA. In order to elucidate the mechanism by which P. copri could contribute to the immunopathogenesis of RA, this project focuses on investigating the effects of P. copri on human monocytes and macrophages. Since it has been reported that patients with active RA have a bias towards proinflammatory M1 type macrophages, it is hypothesized that P. copri reduces viability and promotes the activation and differentiation of monocytes and macrophages towards an M1 profile in vitro. The general objective is to evaluate the effect of P. copri on the viability, activation and differentiation of monocytes and macrophages in vitro. The THP-1 cell line was used as a model for monocytes, which can differentiate into macrophages when exposed to PMA. The effect of P. copri on monocyte and macrophage viability in vitro by using Annexin V labeling and a viability probe by flow cytometry. Then, the effect of P. copri on the activation and differentiation of monocytes and macrophages in vitro was evaluated through the analysis of markers for M1/M2 and cytokine profiling by flow cytometry. This study contributes to the knowledge about the interaction between P. copri and the innate immune system, particularly monocytes and macrophages, in the immunopathogenesis of RA, which could be associated with the development and spread of chronic inflammation in RA.
Notas
Tesis (Licenciada en Biología)
Palabras clave
Artritis Reumatoide, Bacterias Anaeróbicas, Monocitos, Macrófagos