中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2010年
9期
613-616
,共4页
初晓霞%黄葆华%陈丽明%张晓录%于威娟%王彦%王笑蕾%秦友平
初曉霞%黃葆華%陳麗明%張曉錄%于威娟%王彥%王笑蕾%秦友平
초효하%황보화%진려명%장효록%우위연%왕언%왕소뢰%진우평
紫癜,血小板减少性,特发性%树突细胞%Toll样受体%细胞因子
紫癜,血小闆減少性,特髮性%樹突細胞%Toll樣受體%細胞因子
자전,혈소판감소성,특발성%수돌세포%Toll양수체%세포인자
Purpura,thrombocytopenic,idiopathic%Dendritic cells%Toll-like receptor%Cytokines
目的 研究特发性血小板减少性紫癜(ITP)患者树突细胞(DC)免疫表型及其Toll样受体4(TLR4)的表达,探讨其在ITP发病机制中的作用.方法 取ITP完全缓解患者及未达完全缓解患者治疗前后及正常人外周血,分离单个核细胞,加入细胞因子rhGM-CSF及rhIL-4诱导DC,用流式细胞术检测DC表型;酶联免疫吸附法检测DC培养上清液中IL-12p70.应用RT-PCR检测DC的TLR4表达.结果 21例ITP完全缓解患者治疗前DC的CD80和CD86阳性表达率分别为(51.60±13.47)%和(61.50±15.93)%,15例未达完全缓解者CD80和CD86分别为(53.29±19.49)%和(62.91±18.43)%,均高于正常对照的(36.03±15.43)%和(40.28±11.49)%(P<0.01).治疗后ITP缓解患者组CD80和CD86表达率分别降为(36.48±15.19)%和(44.05±17.70)%,与治疗前比较差异有统计学意义(P<0.01),与对照组相比差异无统计学意义(P>0.05);未完全缓解组CD80和CD86分别降为(52.30±20.98)%和(49.79±20.28)%,但与治疗前相比差异均无统计学意义(P>0.05),CD80仍高于正常对照组(P<0.05),而CD86与对照组比较差异亦无统计学意义(P>0.05).地塞米松(DXM)治疗缓解组患者治疗前DC培养上清IL-12p70为(67.52±14.43)pg/ml,明显高于正常对照的(39.78±10.03)pg/ml(P<0.01),治疗后IL-12 p70的表达降至(43.90±8.49)pg/ml,与治疗前比较差异有统计学意义(P<0.01),与正常对照组相比无明显差异;而未缓解组患者DC培养上清IL-12p70则由治疗前的(65.35±12.52)pg/ml降至(48.45±9.68)pg/ml(P<0.01),但仍高于正常对照(P<0.05).治疗缓解组ITP患者DC的TLR4 mRNA相对表达水平为0.69±0.17,明显高于正常对照组的0.31±0.09(P<0.01),治疗后ITP患者DC的TLR4 mRNA相对表达水平降为0.35±0.11(P<0.01),与正常对照组相比无明显差别;未缓解组DC的TLR4 mRNA相对表达水平由治疗前的0.65±0.09降至治疗后的0.52±0.21(P<0.01),但后者仍高于正常对照组(P<0.01).结论 DC可能通过其Toll样受体及细胞因子的分泌在ITP发病中起重要作用.
目的 研究特髮性血小闆減少性紫癜(ITP)患者樹突細胞(DC)免疫錶型及其Toll樣受體4(TLR4)的錶達,探討其在ITP髮病機製中的作用.方法 取ITP完全緩解患者及未達完全緩解患者治療前後及正常人外週血,分離單箇覈細胞,加入細胞因子rhGM-CSF及rhIL-4誘導DC,用流式細胞術檢測DC錶型;酶聯免疫吸附法檢測DC培養上清液中IL-12p70.應用RT-PCR檢測DC的TLR4錶達.結果 21例ITP完全緩解患者治療前DC的CD80和CD86暘性錶達率分彆為(51.60±13.47)%和(61.50±15.93)%,15例未達完全緩解者CD80和CD86分彆為(53.29±19.49)%和(62.91±18.43)%,均高于正常對照的(36.03±15.43)%和(40.28±11.49)%(P<0.01).治療後ITP緩解患者組CD80和CD86錶達率分彆降為(36.48±15.19)%和(44.05±17.70)%,與治療前比較差異有統計學意義(P<0.01),與對照組相比差異無統計學意義(P>0.05);未完全緩解組CD80和CD86分彆降為(52.30±20.98)%和(49.79±20.28)%,但與治療前相比差異均無統計學意義(P>0.05),CD80仍高于正常對照組(P<0.05),而CD86與對照組比較差異亦無統計學意義(P>0.05).地塞米鬆(DXM)治療緩解組患者治療前DC培養上清IL-12p70為(67.52±14.43)pg/ml,明顯高于正常對照的(39.78±10.03)pg/ml(P<0.01),治療後IL-12 p70的錶達降至(43.90±8.49)pg/ml,與治療前比較差異有統計學意義(P<0.01),與正常對照組相比無明顯差異;而未緩解組患者DC培養上清IL-12p70則由治療前的(65.35±12.52)pg/ml降至(48.45±9.68)pg/ml(P<0.01),但仍高于正常對照(P<0.05).治療緩解組ITP患者DC的TLR4 mRNA相對錶達水平為0.69±0.17,明顯高于正常對照組的0.31±0.09(P<0.01),治療後ITP患者DC的TLR4 mRNA相對錶達水平降為0.35±0.11(P<0.01),與正常對照組相比無明顯差彆;未緩解組DC的TLR4 mRNA相對錶達水平由治療前的0.65±0.09降至治療後的0.52±0.21(P<0.01),但後者仍高于正常對照組(P<0.01).結論 DC可能通過其Toll樣受體及細胞因子的分泌在ITP髮病中起重要作用.
목적 연구특발성혈소판감소성자전(ITP)환자수돌세포(DC)면역표형급기Toll양수체4(TLR4)적표체,탐토기재ITP발병궤제중적작용.방법 취ITP완전완해환자급미체완전완해환자치료전후급정상인외주혈,분리단개핵세포,가입세포인자rhGM-CSF급rhIL-4유도DC,용류식세포술검측DC표형;매련면역흡부법검측DC배양상청액중IL-12p70.응용RT-PCR검측DC적TLR4표체.결과 21례ITP완전완해환자치료전DC적CD80화CD86양성표체솔분별위(51.60±13.47)%화(61.50±15.93)%,15례미체완전완해자CD80화CD86분별위(53.29±19.49)%화(62.91±18.43)%,균고우정상대조적(36.03±15.43)%화(40.28±11.49)%(P<0.01).치료후ITP완해환자조CD80화CD86표체솔분별강위(36.48±15.19)%화(44.05±17.70)%,여치료전비교차이유통계학의의(P<0.01),여대조조상비차이무통계학의의(P>0.05);미완전완해조CD80화CD86분별강위(52.30±20.98)%화(49.79±20.28)%,단여치료전상비차이균무통계학의의(P>0.05),CD80잉고우정상대조조(P<0.05),이CD86여대조조비교차이역무통계학의의(P>0.05).지새미송(DXM)치료완해조환자치료전DC배양상청IL-12p70위(67.52±14.43)pg/ml,명현고우정상대조적(39.78±10.03)pg/ml(P<0.01),치료후IL-12 p70적표체강지(43.90±8.49)pg/ml,여치료전비교차이유통계학의의(P<0.01),여정상대조조상비무명현차이;이미완해조환자DC배양상청IL-12p70칙유치료전적(65.35±12.52)pg/ml강지(48.45±9.68)pg/ml(P<0.01),단잉고우정상대조(P<0.05).치료완해조ITP환자DC적TLR4 mRNA상대표체수평위0.69±0.17,명현고우정상대조조적0.31±0.09(P<0.01),치료후ITP환자DC적TLR4 mRNA상대표체수평강위0.35±0.11(P<0.01),여정상대조조상비무명현차별;미완해조DC적TLR4 mRNA상대표체수평유치료전적0.65±0.09강지치료후적0.52±0.21(P<0.01),단후자잉고우정상대조조(P<0.01).결론 DC가능통과기Toll양수체급세포인자적분비재ITP발병중기중요작용.
Objective To study the surface antigen of the dendritic cells(DC) and their Toll-like receptor 4 (TL,R4) expression in patients with idiopathic thrombocytopenic purpura( ITP), and to explore their role in ITP pathogenesis. Methods The peripheral blood mononuclear cells isolated from complete remission patients (CR), non- complete remission patients (n-CR) and normal controls were stimulated by rhGM-CSF and rhlL-4. The surface antigen of the DC was analyzed by flow cytometry. The level of IL-12p70 in the supernatant was detected by enzyme linked immunosorbent assay. The expression of TLR4 mRNA of DC was detected by real time PCR. Results In the 21 CR ITP patients, the expression of both CD80 and CD86 in DC was significantly increased compared with that in normal controls [(51.60 ± 13.47)% vs (36. 03 ±15.43)%, (61.50 ± 15.93)% vs (40.28 ± 11.49)%, respectively] (P<0.01). The expression of CD80and CD86 in n-CR group was also significantly increased [(53.29 ± 19.49)% and (62.91 ± 18.43)%, respectively] (P < 0.01 ). After HD-DXM treatment, both CD80 and CD86 in CR patients were decreased (P<0.01). There was no difference between the DXM treatment patients and the normal controls. In n-CR group, there was no difference in CD80 and CD86 expression before and after DXM therapy [( 52.30 ±20.98% and (49.79 ±20.28)%, respectively] (P>0.05). CD80 was still higher than normal(P <0.05),while CD86 was not changed. The level of IL-12p70 in CR ITP patients before treatment was significantly higher [(67.52 ± 14.43 ) pg/ml] than that of the controls [(39.78 ± 10.03 ) pg/ml] ( P < 0.01 ), and after treatment, was significantly decreased to (43.90 ±8.49) pg/ml, being no difference from that in control. In n-CR group, IL-12p70 was lower after treatment [( 48.45 ± 9.68 ) pg/ml] than that before treatment [(65.35 ±12.52) pg/ml] (P<0.01), but still higher than that in control(P <0.05). The TLR4 mRNA level in DCs of CR ITP patients before treatment were significantly higher 0.69 ±0.17 than that of controls (0.31 ± 0.09 ) ( P < 0. 01 ) and after treatment, was reduced to 0. 35 ± 0. 11, being no difference from that in control. In n-CR group, TLR4 mRNA was decreased from 0. 65 ± 0. 09 to 0.52 ± 0.21 after treatment ( P <0.01 ), but still higher than normal( P <0.01 ). Conclusion DC may play an important role in ITP by their Toll-like receptor and cytokine secretion.