1.Macrophage migration inhibitory factor potentially enhance the proliferation of fibroblast induced by lipopolysaccharide through increasing Toll-like recetor 4
Changyi XIE ; Yebin XI ; Zhengde XI
Chinese Journal of Rheumatology 2014;18(11):766-769,后插2
Objective Based on the research of the mechanism of fibroblast proliferation,the principle of synovial hyperplasia is explored in rheumatoid arthritis (RA).Methods CCK-8 method was used to observe the stimulation effect of LPS on fibroblast proliferation.By confocal scanning microscopy,the expression of TLR-4 on fibroblast membrane was observed; Real-time fluorescent quantitative PCR method was used to detect the stimulatory effect of macrophage migration inhibitory factor on TLR-4 expression of fibrob-lasts and further analyzed the changes of TLR-4 expression on the cell membrane by flow cytometry.Results The LPS could stimulate fibroblast proliferation,and the fibroblasts expression of TLR4 per se.After stimulated by macrophage migration inhibitory factor,the expression of fibroblast TLR-4 was increased by about 20%-30%.Conclusion MIF increases the TLR-4 expression of fibroblasts that potentially enhance cell proliferation induced by LPS.
2.Study on a type Ⅱ collagen-specific T cell line and the pathogenesis of arthritis
Zhengde XI ; Li WANG ; Jun BAI ; Jiying ZHANG ; Baihua SHEN ; Qiwen YU ; Yanyun ZHANG ; Dongqing ZHANG ;
Chinese Journal of Rheumatology 2003;0(09):-
Objective To establish the type Ⅱ collagen specific T cell line of Wistar rat and observe its effect on transferring arthritis.Methods The Wistar rats were immunized with emulsified chicken type Ⅱ collagen (CCⅡ) in complete Freund′s adjuvant by intradermal injection to induce the rat model of collagen induced arthritis (CIA).The lymphocytes were obtained from mesenteric lymph nodes of CIA rats,and the type Ⅱ collagen reactive T cell line was selected and propagated by CCⅡ stimulating in vitro .The proliferation response and phenotype were analyzed by 3 H TdR incorporation and fluorescence activated cell sorter (FACS).The onset of arthritis and pathological characteristic in ankle joints of recipient rats were observed with naked eye and histochemical examination.Anti CCⅡ antibody in serum was assayed by enzyme linked immunosorbent assay (ELISA).Results A T cell line was successfully established.The results of FACS labeled with fluorescent antibodies showed that 98 2% of the line cells were T cells,of which 89 7% were CD4 + T cells.The results of adoptive transfer showed that the incidence of arthritis was 50% when the injected cell number was 5?10 7,meanwhile the level of anti CCⅡ antibody in serum was elevated more than that of the control.Conclusion A cell line has been successfully established.The result of arthritis transferring by T cell line shows that the T cell plays a great role in the pathogenesis of CIA and provides a research datum for rheumatoid arthritis therapy with T cell vaccine.