RESUMO
Objective To investigate the effect of protamine(PS)coated polylactic acid(PLA)nanocapsule on the immune response of murine bone marrow-derived dendritic cells(BMDCs).Methods PLA nanocapsules were prepared by W1/O/W2 method.Protamine was used to modify the surface of PLA nanocapsule that encapsulated ovalbumin(OVA),a model anti-gen.The prepared OVA/PLA/PS nanocapsules were used to stimulate BMDCs.Flow cytometry was used to analyze the ex-pression of surface molecules CD80,CD83,CD86,MHCⅠ,and MHC Ⅱ,and the uptake of nanocapsule.The levels of IL-12 p70 secreted by BMDCs were detected by ELISA.Results OVA/PLA/PS nanocapsules could significantly up-regulate the expres-sion of CD80,CD83,MHCⅠ,and MHC Ⅱ in BMDCs,and increase the secretion of IL-12 p70 by BMDCs.Furthermore,OVA/PLA/PS nanocapsules could enhance the uptake efficiency of OVA by BMDCs.Conclusion OVA/PLA/PS nanocapsule can en-hance the immune response of BMDCs,and may become a good drug delivery carrier.
RESUMO
Objective:To investigate the effect of IL-17A on the differentiation and maturation of murine bone marrow-derived dendritic cells( BMDCs ) . Methods: Murine bone marrow cells were isolated and cultured in RPMI1640 complete medium in the presence of GM-CSF(20 ng/ml) for 8 days to induce differentiation of murine bone marrow cells to DC progenitors. Then these cells were treated with LPS(1 μg/ml) for 36 h which polarized immature DCs into mature DCs. Different concentrations of rmIL-17A(10 or 100 ng/ml) was added to the culture medium at different stages of BMDC differentiation and maturation. Co-stimulatory molecules expression on BMDC were analyzed by flow cytometry,and the culture supernatants were analyzed for IL-12p40 and IL-10 level by ELISA. Results:rmIL-17 could promote co-stimulatory molecules( CD40,CD80,CD86 and MHCⅡ) expression on BMDCs in a does-dependent manner,especially,the expression of CD40 and MHCⅡhad a significant increase in high concentration of rmIL-17A group;rmIL-17A was added while LPS induced maturation of BMDCs. CD40,CD80,CD86 and MHCⅡexpression on BMDC increased sharply in LPS plus rmIL-17A stimulation group,besides,CD86,MHCⅡ showed a higher level expression on BMDC with the increase of con-centration of rmIL-17A. Furthermore,secretion of IL-12p40 and IL-10 increased significantly in the group of DCs treated with LPS plus low concentration of rmIL-17 compared with the group without rmIL-17(P<0. 001). However,high concentration of rmIL-17A group showed significantly higher levels of IL-12p40(P<0. 001),but there was no difference in IL-10. Conclusion:IL-17A promotes the phe-notypic development of BMDC progenitors propagated in GM-CSF and cooperate with LPS to induce BMDC differentiation and matura-tion.