目的 探讨Th17/Treg细胞失衡在儿童再生障碍性贫血(AA)免疫发病及治疗过程中的意义.方法 选取2012年1月至2013年10月在安徽医科大学第二附属医院收治的AA患儿43例(其中男14例),年龄2～ 14岁,根据AA患儿初诊时疾病严重程度分为2组:重型AA(SAA,25例,其中男8例,年龄2 ～14岁)和非重型AA(NSAA,18例,其中男6例,年龄2～14岁).随着治疗的进行,根据临床实际情况制定3个监测时间点:初诊(43例,其中男14例,年龄2～14岁),脱离输血(8例,其中男5例,年龄2 ～11岁)和完全缓解(6例,其中男3例,年龄2～11岁),以同期治疗无效的AA患儿作为无效对照(脱离输血无效对照,5例,其中男1例,年龄3～8岁;完全缓解无效对照,4例,其中男2例,年龄4～11岁).采用流式细胞术检测各研究对象外周血中Treg及Th17细胞占CD4+T细胞的比例,并计算Th 17/Treg;酶联免疫吸附试验(ELISA法)检测初诊AA患儿血浆中白细胞介素(IL)-17和IL-6水平.并检测本院同期25名(其中男12名,年龄3～ 14岁)健康体检儿童外周血中Treg及Th17细胞比例及血浆中IL-17和IL-6水平作为健康对照,以同期9例(其中男4例,年龄1～11岁)化疗后AA状态患儿各项指标作为初诊对照组.采用t检验或ANOVA分析及Mann-Whitney U或Kruskal-WallisH检验进行组间比较;相关性分析采用Spearman相关系数分析.结果 (1)初诊AA患儿外周血中Th17细胞比例1.63％(1.27％,2.48％)高于健康对照组0.40％ (0.35％,0.51％)和初诊对照组0.50％ (0.45％,0.75％);Treg细胞比例4.24％ (3.10％,5.29％)低于正常对照组7.03％ (6.56％,7.48％)和初诊对照组7.50％(6.60％,8.30％);同时Th17/Treg 0.53 (0.34,0.69)高于正常对照组0.06(0.05,0.07)和初诊对照组0.09(0.08,0.11);差异均有统计学意义(P均＜0.01).(2)初诊AA患儿血浆中的IL-17和IL-6均高于正常对照组[(223 ±92)比(116±18)ng/L,(26.2±12.0)比(10.6±2.1)ng/L,P均＜0.01].初诊AA患儿IL-17和IL-6水平均与Th17细胞比例成正相关(r =0.62、0.64,P均＜0.01).(3)SAA患儿外周血中Th17细胞比例、Th17/Treg、IL-17及IL-6均高于正常对照组[1.80％(1.25％,2.61％)比0.40％ (0.35％,0.51％),0.57％ (5.10％,0.82％)比0.06％ (0.05％,0.07％),(225±108)比(116±18) ng/L,(25.9±12.6)比(10.6±2.1)ng/L,P均＜0.01];NSAA患儿外周血中Th17细胞比例、Th17/Treg、IL-17及IL-6均高于正常对照组,但SAA及NSAA患儿的Treg细胞比例均低于正常对照组(P均＜0.01).SAA与NSAA患儿之间的Th17及Treg细胞比例、Th 17/Treg、IL-17及IL-6水平差异均无统计学意义(P均＞0.05).(4)不同治疗阶段(初诊、脱离输血、完全缓解、脱离输血无效对照、完全缓解无效对照)AA患儿Th17细胞比例和Th 17/Treg差异均有统计学意义(P均＜0.05),Treg比例差异均无统计学意义(P均＞0.05).结论 AA患儿外周血中存在Th 17/Treg细胞失衡及TH17细胞相关因子异常增多现象,但对初诊AA的严重程度无明显影响.经免疫抑制治疗后,随着Th17/Treg失衡的纠正,AA得到逐步缓解.
目的 探討Th17/Treg細胞失衡在兒童再生障礙性貧血(AA)免疫髮病及治療過程中的意義.方法 選取2012年1月至2013年10月在安徽醫科大學第二附屬醫院收治的AA患兒43例(其中男14例),年齡2～ 14歲,根據AA患兒初診時疾病嚴重程度分為2組:重型AA(SAA,25例,其中男8例,年齡2 ～14歲)和非重型AA(NSAA,18例,其中男6例,年齡2～14歲).隨著治療的進行,根據臨床實際情況製定3箇鑑測時間點:初診(43例,其中男14例,年齡2～14歲),脫離輸血(8例,其中男5例,年齡2 ～11歲)和完全緩解(6例,其中男3例,年齡2～11歲),以同期治療無效的AA患兒作為無效對照(脫離輸血無效對照,5例,其中男1例,年齡3～8歲;完全緩解無效對照,4例,其中男2例,年齡4～11歲).採用流式細胞術檢測各研究對象外週血中Treg及Th17細胞佔CD4+T細胞的比例,併計算Th 17/Treg;酶聯免疫吸附試驗(ELISA法)檢測初診AA患兒血漿中白細胞介素(IL)-17和IL-6水平.併檢測本院同期25名(其中男12名,年齡3～ 14歲)健康體檢兒童外週血中Treg及Th17細胞比例及血漿中IL-17和IL-6水平作為健康對照,以同期9例(其中男4例,年齡1～11歲)化療後AA狀態患兒各項指標作為初診對照組.採用t檢驗或ANOVA分析及Mann-Whitney U或Kruskal-WallisH檢驗進行組間比較;相關性分析採用Spearman相關繫數分析.結果 (1)初診AA患兒外週血中Th17細胞比例1.63％(1.27％,2.48％)高于健康對照組0.40％ (0.35％,0.51％)和初診對照組0.50％ (0.45％,0.75％);Treg細胞比例4.24％ (3.10％,5.29％)低于正常對照組7.03％ (6.56％,7.48％)和初診對照組7.50％(6.60％,8.30％);同時Th17/Treg 0.53 (0.34,0.69)高于正常對照組0.06(0.05,0.07)和初診對照組0.09(0.08,0.11);差異均有統計學意義(P均＜0.01).(2)初診AA患兒血漿中的IL-17和IL-6均高于正常對照組[(223 ±92)比(116±18)ng/L,(26.2±12.0)比(10.6±2.1)ng/L,P均＜0.01].初診AA患兒IL-17和IL-6水平均與Th17細胞比例成正相關(r =0.62、0.64,P均＜0.01).(3)SAA患兒外週血中Th17細胞比例、Th17/Treg、IL-17及IL-6均高于正常對照組[1.80％(1.25％,2.61％)比0.40％ (0.35％,0.51％),0.57％ (5.10％,0.82％)比0.06％ (0.05％,0.07％),(225±108)比(116±18) ng/L,(25.9±12.6)比(10.6±2.1)ng/L,P均＜0.01];NSAA患兒外週血中Th17細胞比例、Th17/Treg、IL-17及IL-6均高于正常對照組,但SAA及NSAA患兒的Treg細胞比例均低于正常對照組(P均＜0.01).SAA與NSAA患兒之間的Th17及Treg細胞比例、Th 17/Treg、IL-17及IL-6水平差異均無統計學意義(P均＞0.05).(4)不同治療階段(初診、脫離輸血、完全緩解、脫離輸血無效對照、完全緩解無效對照)AA患兒Th17細胞比例和Th 17/Treg差異均有統計學意義(P均＜0.05),Treg比例差異均無統計學意義(P均＞0.05).結論 AA患兒外週血中存在Th 17/Treg細胞失衡及TH17細胞相關因子異常增多現象,但對初診AA的嚴重程度無明顯影響.經免疫抑製治療後,隨著Th17/Treg失衡的糾正,AA得到逐步緩解.
목적 탐토Th17/Treg세포실형재인동재생장애성빈혈(AA)면역발병급치료과정중적의의.방법 선취2012년1월지2013년10월재안휘의과대학제이부속의원수치적AA환인43례(기중남14례),년령2～ 14세,근거AA환인초진시질병엄중정도분위2조:중형AA(SAA,25례,기중남8례,년령2 ～14세)화비중형AA(NSAA,18례,기중남6례,년령2～14세).수착치료적진행,근거림상실제정황제정3개감측시간점:초진(43례,기중남14례,년령2～14세),탈리수혈(8례,기중남5례,년령2 ～11세)화완전완해(6례,기중남3례,년령2～11세),이동기치료무효적AA환인작위무효대조(탈리수혈무효대조,5례,기중남1례,년령3～8세;완전완해무효대조,4례,기중남2례,년령4～11세).채용류식세포술검측각연구대상외주혈중Treg급Th17세포점CD4+T세포적비례,병계산Th 17/Treg;매련면역흡부시험(ELISA법)검측초진AA환인혈장중백세포개소(IL)-17화IL-6수평.병검측본원동기25명(기중남12명,년령3～ 14세)건강체검인동외주혈중Treg급Th17세포비례급혈장중IL-17화IL-6수평작위건강대조,이동기9례(기중남4례,년령1～11세)화료후AA상태환인각항지표작위초진대조조.채용t검험혹ANOVA분석급Mann-Whitney U혹Kruskal-WallisH검험진행조간비교;상관성분석채용Spearman상관계수분석.결과 (1)초진AA환인외주혈중Th17세포비례1.63％(1.27％,2.48％)고우건강대조조0.40％ (0.35％,0.51％)화초진대조조0.50％ (0.45％,0.75％);Treg세포비례4.24％ (3.10％,5.29％)저우정상대조조7.03％ (6.56％,7.48％)화초진대조조7.50％(6.60％,8.30％);동시Th17/Treg 0.53 (0.34,0.69)고우정상대조조0.06(0.05,0.07)화초진대조조0.09(0.08,0.11);차이균유통계학의의(P균＜0.01).(2)초진AA환인혈장중적IL-17화IL-6균고우정상대조조[(223 ±92)비(116±18)ng/L,(26.2±12.0)비(10.6±2.1)ng/L,P균＜0.01].초진AA환인IL-17화IL-6수평균여Th17세포비례성정상관(r =0.62、0.64,P균＜0.01).(3)SAA환인외주혈중Th17세포비례、Th17/Treg、IL-17급IL-6균고우정상대조조[1.80％(1.25％,2.61％)비0.40％ (0.35％,0.51％),0.57％ (5.10％,0.82％)비0.06％ (0.05％,0.07％),(225±108)비(116±18) ng/L,(25.9±12.6)비(10.6±2.1)ng/L,P균＜0.01];NSAA환인외주혈중Th17세포비례、Th17/Treg、IL-17급IL-6균고우정상대조조,단SAA급NSAA환인적Treg세포비례균저우정상대조조(P균＜0.01).SAA여NSAA환인지간적Th17급Treg세포비례、Th 17/Treg、IL-17급IL-6수평차이균무통계학의의(P균＞0.05).(4)불동치료계단(초진、탈리수혈、완전완해、탈리수혈무효대조、완전완해무효대조)AA환인Th17세포비례화Th 17/Treg차이균유통계학의의(P균＜0.05),Treg비례차이균무통계학의의(P균＞0.05).결론 AA환인외주혈중존재Th 17/Treg세포실형급TH17세포상관인자이상증다현상,단대초진AA적엄중정도무명현영향.경면역억제치료후,수착Th17/Treg실형적규정,AA득도축보완해.
Objective To study the role of Th17/Treg imbalance in the immune pathogenesis and therapeutic significance in childhood aplastic anemia (AA).Method We analyzed data from 43 children (male∶female =14∶ 29) with AA,all the cases were at the age of 2 to 14 years at diagnosis,and were hospitalized at our department of pediatrics between January 2012 and October 2013 in the Second Hospital of Anhui Medical University.All these patients were divided into 2 groups,severe AA (SAA) group (n =25,male∶ female =8∶ 17,2-14 years old) and non-severe AA (NSAA) group (n =18,male∶ female =6∶ 12,2-14 years old),depending on the severity at first diagnosis.As to the treatment,we analyzed data at 3 phases of treatment,diagnosis (n =43,male ∶ female =14 ∶ 29,2-14 years old),transfusionindenpendence (n =8,male∶ female =5∶3,2-11 years old),complete response (n =6,male∶ female =3∶3,2-11 years old); at the same time,AA children who did not respond to the treatments were considered as failed treatment control (transfusion-indenpendence with failed treatment group,n =5,male∶ female =1∶4,3-8 years old; complete response failed treatment group,n =4,male∶ female =2∶ 2,4-11 years old).The ratio of Treg and Th17 cells in CD4+ T cells were tested by flow cytometry.The levels of IL-6 and IL-17 in plasma were determined by ELISA.During the same period,25 age-matched healthy children (male∶ female =12∶ 13,3-14 years old) were recruited as normal control,9 cases (male∶ female =5∶3,2-11 years old) of AA children induced by chemotherapy as diagnosis control group.Differences in variables were analyzed using ANOVA and t-tests or the Kruskal-Wallis and Mann-Whitney Utests,as appropriate.Correlation analysis was evaluated by the Spearman rank correlation test.Result (1) The ratio of Th17 cells in newly diagnosed AA patients were higher than that of normal group or diagnosis control group [1.63％(1.27％,2.48％) vs.0.4％ (0.35％,0.51％) or 0.50％ (0.45％,0.75％),both P ＜0.01] while the ratio of Treg cells was lower [4.24％ (3.10％,5.29％) vs.7.03％ (6.56％,7.48％) or 7.50％ (6.60％,8.30％),both P ＜0.01] and the proportion of Th17/Treg were significantly higher [0.53 (0.34,0.69) vs.0.06 (0.05,0.07) or 0.09 (0.08,0.11),both P ＜ 0.01].(2) The levels of IL-6 and IL-17 in newly diagnosed AA patients were higher than in normal group [(223 ± 92) vs.(116 ± 18) ng/L,(26.2 ± 12.0) ng/L vs.(10.6 ±2.1) ng/L,P both ＜0.01].There was a positive correlation between Th17 cells and some Th17 cells related cytokines such as IL-17 and IL-6 (r =0.62,0.64,P both ＜0.01).(3) The ratio of Th17,Th17/Treg,and the levels of IL-6 and IL-17 in children with SAA were also higher than in normal group [1.80％ (1.25％,2.61％) vs.0.40％ (0.35％,0.51％),0.57％(5.10％,0.82％) vs.0.06％ (0.05％,0.07％),(225±108) vs.(116±18) ng/L,(25.9 ± 12.6) vs.(10.6 ±2.1)ng/L,all P ＜0.01].NSAA also higher than normal group.The ratio of Treg in children with SAA and NSAA was less than that in normal group (P all ＜0.01).However,the ratio of Th17,Treg,Th17/Treg,and the levels of IL-6 and IL-17 had no significant difference between SAA and NSAA (all P ＞ 0.05).(4) In different stages of treatment,such as diagnosis,transfusion-indenpendence,complete response,there were significant differences in the ratio of Th17 and Th17/Treg (both P ＜ 0.05) but not in Treg (P ＞ 0.05).Conclusion The imbalance of Th17/Treg cells and abnormally increased cytokines related to Th17 cells exist in peripheral blood of AA children,but did not significantly affect the severity of AA in preliminary diagnosis.After treatment with immunosuppression,AA was gradually relieved as the imbalance of Th17/Treg was corrected.
