CAJ | 학술논문

目的研究不同激活状态下T辅助细胞(Th)中IL-2R各亚基的表达变化及对IL-2反应的相关性.方法以包被的抗CD3抗体激活Th1细胞,同时设立未激活对照.3H掺入法测定其对IL-2的促增殖反应；real-time PCR检测IL-2R各亚基编码基因的表达；流式细胞术检测CD25及CD122的含量；以I125标记的IL-2检测不同状态的Th1细胞对IL-2的亲和力.脂质体法转染IL-2Rα siRNA至激活的Th1细胞,比较不同CD25含量时Th1细胞对IL-2的反应性.自小鼠体内分离CD4细胞,以抗CD3抗体激活后在不同时间点收集细胞,比较它们的CD25含量及对1L-2的反应性.结果较未激活对照相比,激活的Th1细胞中IL-2Rα亚基,即CD25的表达显著增加,而其他两个亚基则无变化；同时伴有对IL-2亲和力的升高,但对IL-2的促增殖反应性却显著降低.以siRNA适当下调IL-2Rα基因的表达则可显著提高激活的Th1细胞对IL-2的反应性.虽然激活后的na(i)ve CD4细胞对IL-2的反应性明显增加,但却不与CD25的含量及激活度正相关.最高的IL-2反应性出现在低度激活的CD4细胞中.结论 CD25在激活后的Th细胞中显著增多,并可通过自身数量的变化来调节所在细胞对IL-2的促增殖反应性.适度高表达的CD25可使靶细胞具有最高的IL-2反应性,过度表达的CD25则可导致对IL-2反应性的降低.
목적 연구불동격활상태하T보조세포(Th)중IL-2R각아기적표체변화급대IL-2반응적상관성.방법 이포피적항CD3항체격활Th1세포,동시설립미격활대조.3H참입법측정기대IL-2적촉증식반응；real-time PCR검측IL-2R각아기편마기인적표체；류식세포술검측CD25급CD122적함량；이I125표기적IL-2검측불동상태적Th1세포대IL-2적친화력.지질체법전염IL-2Rα siRNA지격활적Th1세포,비교불동CD25함량시Th1세포대IL-2적반응성.자소서체내분리CD4세포,이항CD3항체격활후재불동시간점수집세포,비교타문적CD25함량급대1L-2적반응성.결과 교미격활대조상비,격활적Th1세포중IL-2Rα아기,즉CD25적표체현저증가,이기타량개아기칙무변화；동시반유대IL-2친화력적승고,단대IL-2적촉증식반응성각현저강저.이siRNA괄당하조IL-2Rα기인적표체칙가현저제고격활적Th1세포대IL-2적반응성.수연격활후적na(i)ve CD4세포대IL-2적반응성명현증가,단각불여CD25적함량급격활도정상관.최고적IL-2반응성출현재저도격활적CD4세포중.결론 CD25재격활후적Th세포중현저증다,병가통과자신수량적변화래조절소재세포대IL-2적촉증식반응성.괄도고표체적CD25가사파세포구유최고적IL-2반응성,과도표체적CD25칙가도치대IL-2반응성적강저.
Objective To elucidate the expression diversity of IL-2R subunits in T helper(Th) cells under different activation condition and clarify its relationship with the response to IL-2 induced proliferation.Methods In vitro cultured Th1 cells were activated by plate bound anti-mouse CD3 McAb.In both activated and inactivated Th1 cells the expression of different IL-2R subunit was evaluated by real-time PCR and fluorescence staining,3H incorporation assay was adopt to measure the proliferation of the cells in response to IL-2 treatment,IL-2 affinity was determined via I125 labeled IL-2.IL-2Rα siRNA transfection was applied to knockdown CD25 expression in activated Th1 cells and its effect was confirmed by Western blot,IL-2 induced proliferation in IL-2Rα siRNA transfected Th1 cells were subsequently detected.CD4 cells were isolated from na(i)ve BALB/c mice and were also stimulated by plate bound anti-CD3 McAb.Cells were harvested at different days posterior to the stimulation,CD25 content and IL-2 induced proliferation were determined by flow cytometry and 3H incorporation assay respectively.Results In comparison with inactivated cells,only the expression of CD25but not other IL-2R subunits was significantly increased in activated Th1 cells.In accordance with elevated CD25,IL-2 affinity was also increased in activated Th1 cells,however,which resulted in decreased cell proliferation in response to IL-2 treatment.In activated Th1 cells,when CD25 expression was differently knockdown by different-dosed IL-2Rα siRNA transfection,the highest IL-2 response was observed in the cells with partially CD25 depression.Forin vitro activated na(i)ve CD4 cells the elevated CD25 expression gradually decreased and the highest proliferation response was detected at day 8 post stimulation upon IL-2 treatment.Conclusion Although CD25 is necessary for IL-2 induced proliferation,however,the excessively expressed CD25 lead to lower proliferation response.Not fully activated Thl or CD4 cells,but partially activated cells with lightly increased CD25 content possessed highest proliferation rate in response to IL-2 treatment.