The effects of tolerogenic dendritic cells on the TLR4/MyD88/NF-κB signaling pathway in rats with collagen-induced arthritis

Abstract

Objective: Rheumatoid arthritis (RA) is a common inflammatory autoimmune disease. Previous studies have emphasized tolerogenic dendritic cells(tolDCs) could attenuate inflammatory lesions by inducing specific immune tolerance in RA animal models, but the mechanism still needs further investigation. This study focused on revealing the effects of tolDCs on the TLR4/MyD88/NF-κB signaling pathway that mediates inflammation.

Methods: Bone marrow-derived tolDCs were induced by IL-4, GM-CSF and NF-κB Oligonucleotide Decoys. The DC-specific molecule OX-62 and co-stimulatory molecules CD80 and CD86 on the surface of tolDCs were detected by flow cytometry. Joint damage was assessed by H&E, Safranine O-fast green staining and tartrate-resistant acid phosphase (TRAP) staining, and the histological change of spleen tissue was also evaluated by H&E staining. Immunohistochemistry (IHC) was performed to detect key proteins of TLR4/MyD88/NF-κB signaling pathway of synovium, cartilage, and bone tissues of ankle joints respectively. Immunofluorescence (IF) was performed to observe NF-κB p65 nuclear translocation and subcellular localization of phosphorylated NF-κB p65 (p-NF-κB p65).

Results: The intervention of tolDCs showed a significant reduction in joint inflammation and destruction in CIA rats. Moreover, tolDCs suppressed the hyperactivation of the TLR4/MyD88/NF-κB signaling pathway of the cells in synovium, cartilage and bone tissues of ankle joints in CIA rats.

Conclusions: TolDCs may exert therapeutic effects on CIA rats by alleviating the inflammation through inhibiting the hyperactivation of the TLR4/MyD88/NF-κB signaling pathway.

Keywords: Collagen-induced arthritis; Inflammation; Rheumatoid arthritis; TLR4/MyD88/NF-κB signaling pathway; Tolerogenic dendritic cells.