胆道闭锁肝内三级淋巴结构与预后的相关性研究  

Correlation between tertiary lymphoid structures with prognosis of children with biliary atresia

作　　者：武少静 马家强 陈功[1] 张晓明[3] Wu Shaojing;Ma Jiaqiang;Chen Gong;Zhang Xiaoming(Department of Surgery,Children's Hospital,Fudan University,Shanghai 201102;Department of Hepatic Surgery,Zhongshan Hospital,Fudan University,Shanghai 200032;Shanghai Institute Pasteur,Chinese Academy of Science,Shanghai 200031)

机构地区：[1]复旦大学附属儿科医院外科,上海201102 [2]复旦大学附属中山医院肝外科,上海200032 [3]中国科学院上海巴斯德研究所,上海200031

出　　处：《中华小儿外科杂志》2022年第10期894-900,共7页Chinese Journal of Pediatric Surgery

摘　　要：目的分析胆道闭锁(biliary atresia,BA)肝内三级淋巴结构(Tertiary Lymphoid structures,TLSs)的数量、类型及内部Treg、Tfh浸润水平与患儿预后的关系。方法选取2015年2月至2017年5月在复旦大学附属儿科医院接受Kasai手术患儿30例(BA组)肝脏标本,根据患儿术后5年的随访情况将其分为:预后良好组(BA-Good,19例)、预后不良组(BA-Poor,11例),7例胆总管囊肿患儿肝脏标本为对照组。肝楔形活检样本行HE染色、免疫荧光染色,观察TLSs的形态特征并通过细胞计数将其分为Ⅰ型,Ⅱ型,Ⅲ型,比较不同组间TLSs总数及分型占总数的百分比,以及TLSs中Treg与Tfh的浸润水平差异。组间进行Mann-Whitney U和Kruskal-wallis检验,绘制ROC曲线,计算曲线下面积,确定最佳临界值。结果①BA组的TLSs总数[9.50(7.75,13.75)]较对照组[6.00(4.00,6.00)]显著增加(P=0.0002);BA组Ⅰ型TLSs的百分比明显降低[(27.53±2.04)%比(49.22±6.18)%,P=0.0022],Ⅲ型TLSs的百分比明显升高[(28.82±2.27)%比(9.18±3.26)%,P=0.0007]。②Ⅲ型TLSs的百分比预后不良组较预后良好组显著升高[(39.25±2.85)%比(22.79±2.22)%,P=0.0020]。③以Ⅲ型TLSs百分比和患儿的预后绘制ROC曲线:AUC为0.93,P=0.0001,36.50%为最佳临界值,敏感度为0.82,特异度为1.00。④与对照组相比,BA组TLSs内部Tfh的浸润水平明显升高[(0.0391±0.0042)%比(0.0063±0.0027)%,P=0.0018],Treg的浸润水平显著降低[(0.0023±0.0014)%比(0.0082±0.0021)%,P<0.0001]。⑤Ⅲ型TLSs的百分比≥36.50%时,Tfh细胞的浸润水平有升高趋势[(0.0469±0.0051)%比(0.0344±0.0032)%,P=0.0524],Treg的浸润水平明显降低[(0.0012±0.0001)%比(0.0016±0.0001)%,P=0.0052]。结论胆道闭锁肝脏中TLSs数目增多,Ⅲ型TLSs的百分比增加提示患儿预后不良;胆道闭锁肝组织TLSs中Tfh浸润水平增加,Treg浸润水平降低,共同促进了TLSs的成熟,与患儿预后不良有关。Objective To explore the prognostic relationship between the number/type of tertiary lymphoid structures(TLSs),the infiltration levels of Treg and Tfh in children with biliary atresia(BA).Methods From February 2015 to May 2017,30 children(BA group)undergoing Kasai surgery at Pediatric Hospital of Fudan University were recruited and divided into two groups of good prognosis(BA-Good,n=19)and poor prognosis(BA-Bad,n=11).Liver specimens from 7 children with choledochal cyst served as controls.Liver wedge biopsy samples were treated with hematoxylin-eosin(HE)and immunofluorescent stain for observing the morphological characteristics of TLSs,and they were classified into typesⅠ,ⅡandⅢby cell counting.Differences in infiltration levels between Treg and Tfh were recorded.Mann-Whitney U and Kruskal-Wallis inter-group tests were performed.Receiver operating characteristic(ROC)curve was plotted,the area under the curve calculated and optimal critical value determined.P<0.05 implied statistical difference.Results Compared with control group,total number of TLSs significantly spiked in BA group[9.50(7.75,13.75)]vs[6.00(4.00,6.00)],P=0.0002;the percentage of type I TLSs dropped markedly in BA group(27.53±2.04%vs 49.22±6.18%,P=0.0022)while the percentage of typeⅢTLSs significantly increased(28.82±2.27%vs 9.18±3.26%,P=0.0007).The percentage of typeⅢTLSs was significantly higher in poor prognosis group than that in good prognosis group(39.25±2.85%vs 22.79±2.22%,P=0.0020).ROC curve was plotted based upon the percentage of typeⅢTLSs and patient prognosis:AUC was 0.93,P=0.0001,optimal cutoff value 36.50%,sensitivity 0.82 and specificity 1.00;Compared with control group,infiltration level of Tfh in TLSs spiked obviously in BA group(0.0391±0.0042%vs 0.0063±0.0027%,P=0.0018)while infiltration level of Treg declined markedly(0.0023±0.0014%vs 0.0082±0.0021%,P<0.0001).With a percentage of typeⅢTLSs≥36.50%,the infiltration level of Tfh spiked(0.0469±0.0051%vs 0.0344±0.0032%,P=0.0524)while the infiltration level o

关 键 词：胆道闭锁 三级淋巴结构 

分 类 号：R726.5[医药卫生—儿科]