Gwendolyne Méndez-Frausto1, Marina Nayeli Medina-Rosales1, Edith Elena Uresti-Rivera2, Lourdes Baranda-Cándido3, Martín Zapata-Zúñiga4, Yadira Bastián5, Roberto González Amaro2, José Antonio Enciso-Moreno1, Mariana Haydee García-Hernández6. 1. Unidad de Investigación Biomédica. Delegación Zacatecas. Instituto Mexicano del Seguro Social, IMSS, C.P. 98000, Mexico. 2. Centro de Investigación en Ciencias de la Salud y Biomedicina. CICSaB Universidad Autónoma de San Luis Potosí, UASLP, C.P. 78000, Mexico. 3. Centro de Investigación en Ciencias de la Salud y Biomedicina. CICSaB Universidad Autónoma de San Luis Potosí, UASLP, C.P. 78000, Mexico; Unidad Regional de Reumatología y Osteoporosis Hospital Central Dr. Ignacio Morones Prieto. San Luis Potosí, SLP, C.P. 78290, Mexico. 4. Facultad de Medicina y Ciencias de la Salud, Universidad Autónoma de Zacatecas, Hospital Rural No. 51 IMSS Bienestar, Villanueva, Zacatecas, C.P. 99559, Mexico. 5. Unidad de Investigación Biomédica. Delegación Zacatecas. Instituto Mexicano del Seguro Social, IMSS, C.P. 98000, Mexico; Cátedras CONACYT- Unidad de Investigación Biomédica de Zacatecas-IMSS, Zacatecas, C.P. 98000, Mexico. 6. Unidad de Investigación Biomédica. Delegación Zacatecas. Instituto Mexicano del Seguro Social, IMSS, C.P. 98000, Mexico. Electronic address: mariana.garciah@imss.gob.mx.
Abstract
INTRODUCTION: AIM2 inflammasome activation leads to the release of IL-β, which plays an important role in rheumatoid arthritis pathogenesis. In this work, we evaluated AIM2 expression and activity in RA patients and healthy controls. METHODS: AIM2 and RANKL expression were evaluated by flow cytometry. Inflammasome activity was determined in monocyte cultures stimulated with synthetic DNA by measuring IL-1β levels in supernatants using an ELISA assay. The caspase-1 expression in monocytes was measured by western blot, the POP3 expression was analysed by qPCR, and serum levels of IFN-γ were evaluated using ELISA assay. RESULTS: We observed a diminution of CD14+AIM2+ cells in RA patients, associated with disease activity and evolution. Likewise, the levels of IL-1β were increased in monocyte cultures un-stimulated and stimulated with LPS from RA patients with DAS28 ≥ 4. The Caspase-1 activity and RANKL + monocytes in RA patients were slightly increased. Finally, augmented POP3 expression and diminished IFN-γ serum levels were detected in RA patients. CONCLUSION: Our results showed that the monocytes from RA patients were prone to release IL-1β in the absence of the AIM2 inflammasome signal. The down-regulation of AIM2 to a systemic level in RA patients might be a consequence of augmented POP3 expression and might imply the survival of pro-inflammatory cells contributing to the inflammation process.
INTRODUCTION:AIM2 inflammasome activation leads to the release of IL-β, which plays an important role in rheumatoid arthritis pathogenesis. In this work, we evaluated AIM2 expression and activity in RApatients and healthy controls. METHODS:AIM2 and RANKL expression were evaluated by flow cytometry. Inflammasome activity was determined in monocyte cultures stimulated with synthetic DNA by measuring IL-1β levels in supernatants using an ELISA assay. The caspase-1 expression in monocytes was measured by western blot, the POP3 expression was analysed by qPCR, and serum levels of IFN-γ were evaluated using ELISA assay. RESULTS: We observed a diminution of CD14+AIM2+ cells in RApatients, associated with disease activity and evolution. Likewise, the levels of IL-1β were increased in monocyte cultures un-stimulated and stimulated with LPS from RApatients with DAS28 ≥ 4. The Caspase-1 activity and RANKL + monocytes in RApatients were slightly increased. Finally, augmented POP3 expression and diminished IFN-γ serum levels were detected in RApatients. CONCLUSION: Our results showed that the monocytes from RApatients were prone to release IL-1β in the absence of the AIM2 inflammasome signal. The down-regulation of AIM2 to a systemic level in RApatients might be a consequence of augmented POP3 expression and might imply the survival of pro-inflammatory cells contributing to the inflammation process.