Abstract
To construct and evaluate rats' tolerogenic dendritic cells (DC) through induction by NF-κB Decoy method. GM-CSF and IL-4 were used to transform rats's monocytes into DC, and DC were stimulated with LPS, NF-κB Decoy ODN, and loaded with Bovine Type II Collagen. The following methods were employed to phenotype DC: 1) Observation of cell morphology; 2) Evaluation of cell viability using trypan blue staining; 3) Purity determination of DC through detection of specific markers OX-62; 4) Evaluation of mature state of DC via the determination of the expression of CD80 and CD86; 5) Determination of stimulation capability towards the proliferation of lymphocyte and the secretion of INF-r and IL-10. The activity of DC was more than 92%, and the expression of OX-62 was more than 70%. Most of DC exhibited the phenotype of CD80(+)/CD86(-). Compared with control group and LPS-stimulation group, the less mature adhered cells and hairlike DC were observed in NF-κB decoy group. Significant reduction (p < 0.05) was observed for the positive expression and extension of CD80 and CD86 in cell surface. After loaded with calf type II collagen, the low expression of CD80 and CD86 remains to be existed. The stimulation capability of DC towards lymphocyte in NF-κB decoy group was lower than that in control group (p<0.05) and LPS stimulation group (p < 0.05). NF-κB Decoy ODN method can be successfully applied for construct rats' tolerogenic dendritic cells (DC) with stable morphology and phenotype. The tolerogenic DC exhibited immature immune phenotype, and low capability to stimulate lymphocytes.
Highlights
Rheumatoid arthritis (RA) is a chronic autoimmune disease influencing multiple joints such as hands, wrists and feet
The transfection of NF-κB Decoy ODN made Dendritic cells (DC) to maintain the following immature state: 1) The adhered and hairlike cells are more in decoy group than control group; 2) In NF-κB Decoy ODN group, the expression of CD80 and CD86 in cell surface is lower than that in control group
LPS stimulation was used to stability of immatured DCs, and the results showed the expression of CD80 and CD86 was not significantly altered, indicating that decoy-induced DC was very stable
Summary
Rheumatoid arthritis (RA) is a chronic autoimmune disease influencing multiple joints such as hands, wrists and feet. The clinical therapeutic drugs for treatment of RA commonly focus on the inhibition of immune system, which provides big risk of infectious diseases. Searching of new therapeutic methods for RA is necessary and important. The initiation of RA has been closely related with the abnormal antigen presenting cell (APC), which will result in the activation of T cells, especially Th1 and Th17 cells [3]. Dendritic cells (DC) is a kind of APC, and play a key role in regulation of the induction of the differentiation naive T cells towards Th cells through increased expression of adhesion molecules and cytokine receptors as well as the production of cytokines [4].
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
