背景:关于流式细胞术(FC)监测难治性乳糜泻(RCD)中上皮内淋巴细胞(IEL)的数据很少,无反应性乳糜泻(NRCD),和非腹腔肠道病变(NCEs)。
目的:1)探讨NRCD患者用FC监测IELs免疫表型的意义,RCD和NCE;2)用免疫组织化学(IHC)和γ-TCR克隆性分析评估FC的一致性。
方法:在2012年1月至2023年2月之间进行调查的患者分为两组:1)确认RCD或NRCD的持续性症状和乳糜泻(CD)的可疑并发症;2)缺乏临床/组织学反应的NCEs。回顾性收集临床/分子特征和结果,并根据FC上是否存在异常IEL(截止值≥20%CD103sCD3-CD8-iCD3IEL)进行分析。
结果:52例患者(18例RCD,21NRCD,13个NCE;38F,55±13年;中位随访30个月,IQR2-58)进行了100次FCIEL测定。22/52的FC测定≥2个,IEL表型随时间保持不变(κ=1.00)。CD中IEL表型异常与死亡率增加相关(HR4.2,95%CI1.5-11.9,p<0.01)。没有NCEs患者在FC有异常的IEL表型,尽管3/13发展为淋巴瘤和4/13死亡。FC的一致性与IHC(κ=0.40)和γ-TCR克隆性分析(κ=0.22)均相当。
结论:FC可准确评估和监测IEL表型,并为乳糜泻患者提供重要的预后信息。需要进一步研究其在NCEs中的作用。
BACKGROUND: Few data are available on flow cytometry (FC) for monitoring intraepithelial lymphocytes (IELs) in refractory celiac disease (RCD), non-responsive celiac disease (NRCD), and non-celiac enteropathies (NCEs).
OBJECTIVE: 1) To investigate the significance of monitoring IELs immunophenotype with FC in patients with NRCD, RCD and NCEs; 2) to evaluate FC concordance with immunohistochemistry (IHC) and γ-TCR clonality analysis.
METHODS: Patients investigated between January-2012 and February-2023 were divided into two groups: 1)confirmed RCD or NRCD being investigated for persistent symptoms and suspected complications of celiac disease (CD); 2)NCEs lacking clinical/histological response. Clinical/molecular features and outcomes were retrospectively collected and analysed according to presence/absence of aberrant IELs on FC (cut-off≥20 % CD103+sCD3-CD8-iCD3+ IELs).
RESULTS: 52 patients (18 RCD,21 NRCD,13 NCEs; 38F, 55±13 years; median follow-up 30 months, IQR 2-58) underwent 100 FC IELs determinations. 22/52 had ≥2 FC determinations and IEL phenotype remained unchanged over time in all them (κ=1.00). Aberrant IEL phenotype in CD was associated with increased mortality (HR 4.2, 95 % CI 1.5-11.9, p < 0.01). No patients with NCEs had an aberrant IEL phenotype at FC, although 3/13 developed lymphoma and 4/13 died. Concordance of FC was fair with both IHC (κ=0.40) and γ-TCR clonality analysis (κ=0.22).
CONCLUSIONS: FC is accurate for assessing and monitoring IEL phenotype and providing important prognostic information in celiac patients. Further study is needed on its role in NCEs.