RESUMO
Objective:To investigate effects of a novel synthetic immunostimulator CH1b containing thiazolidin-4-one on the immunoregulation funotion of iNKT ( invariant nature killer T ) cells in active RA patients in vitro.Methods: Peripheral blood mononuclear cells( PBMCs) isolated from active RA patients were cultured with stimulation of α-Galcer and IL-2 in vitro and iNKT cells were then separated by using magnetic activated cell sorting( MACS) method with iNKT isolation kit.The cells were divided into three groups:control group (IL-2),α-Galcer group (IL-2+α-Galcer),CH1b group(IL-2 +CH1b).The effects of CH1b on the proliferation of iNKT cells in active RA patients were analyzed by using MTT assay.MILLIPLEX MAP Human Cytokine/Chemokine kit was used to evaluate the secretion of IFN-γand IL-4 in iNKT cells culture media.The expressions of IFN-γmRNA and IL-4 mRNA in iNKT cells were analyzed by RT-PCR.Results: Compared with control and α-Galcer group,the proliferation of iNKT cells of CH1b group were significantly higher( P<0.05).Compared with control,the ratio of IFN-γ/IL-4 in iNKT cells culture media in active RA patients of CH1b group were significantly lower (P<0.05).Compared with control,expressions of IFN-γmRNA and IL-4 mRNA were higher inα-Galcer group;compared with control,expressions of IL-4 mRNA were higher in CH1b group,while there were no obvious difference on expressions of IFN-γmRNA.Conclusion:CH1b was found to significantly stimulate the actived iNKT cells in active RA patients proliferation,promote the secretion of IL-4,and increase the ratio of IFN-γ/IL-4,promote the expression of IL-4 mRNA in iNKT cells in active patients.
RESUMO
Objective To investigate the alterations of invariant nature killer T( iNKT) cells in peripheral blood samples from patients with rheumatoid arthritis ( RA) and to clarify the correlation between the percentage of iNKT cells and the ratio of IFN-γ/IL-4 in order to further understand the significance of iNKT cells in the development of RA.Methods Peripheral blood mononuclear cells ( PBMCs) were isola-ted from 70 patients with RA and 40 healthy subjects.Among them, thirty patients in the stage of inactive RA were involved in a follow-up study.Fluorescence activated cell sorting ( FACS) was used to detect the percentage of iNKT cells.PBMCs were cultured in vitro for analysis of cytokine production.The dynamic changes of iNKT cells in percentages were analyzed by FACS.MILLIPLEX MAP Human Cytokine/Chemo-kine kit was used to measure the secretion of IFN-γand IL-4 in serum samples and culture media of PBMCs. The expression of IFN-γand IL-4 in iNKT cells at mRNA level were analyzed by RT-PCR.Results Com-pared with the healthy subjects, the patients with active RA showed the delayed proliferation of iNKT cells and the decreased percentages and proliferation rates of iNKT cells (P0.05).The ratios of IFN-γ/IL-4 in serum samples and culture media of PBMCs were increased in patients with active RA as compared with those in patients with inactive RA and healthy subjects (P0.05).Compared with healthy subjects and patients with inactive RA, patients with active RA showed increased transcriptional level of IFN-γand decreased transcriptional level of IL-4.No significant differences with the expression of IFN-γand IL-4 in iNKT cells at mRNA level were observed between healthy subjects and patients with inactive RA.The per-centage of iNKT cells was negatively related to the IFN-γ/IL-4 ratio in patients with RA (P<0.05).Con-clusion Decreased percentage and impaired function of iNKT cells were detected in patients with RA. iNKT cells were closely related to the development and disease activity of RA.