Mandile Roberta, Maglio Mariantonia, Marano Antonella, Rapacciuolo Luciano, Discepolo Valentina, Troncone Riccardo, Auricchio Renata
European Laboratory for the Investigation of Food Induced Diseases (ELFID) University of Naples Federico II, Naples, Italy.
Department of Translational Medical Sciences, Section of Paediatrics, University of Naples Federico II, Naples, Italy.
APMIS. 2025 Mar;133(3):e70015. doi: 10.1111/apm.70015.
Intraepithelial lymphocytes (IELs) count, central for coeliac disease (CD) diagnosis, can be performed either directly on hematoxylin and eosin (H&E)-stained paraffined sections or on optimal-cutting-temperature-compound (OCT)-embedded frozen sections stained by immunohistochemistry (IHC) with anti-CD3. We evaluated the concordance in Marsh grading between these two techniques on a large sample of sections. A total of 280 patients with a normal intestinal architecture, 210 potential celiac disease (PCD) patients, and 70 controls (CTR) were included. At the H&E histological evaluation, 136/280 were classified as Marsh-0 (showing < 25 IELs/100 enterocytes) and 144 Marsh-1, while at the IHC evaluation, 191 were classified as Marsh-0 (showing ≤ 34 CD3+/mm of epithelium) and 89 Marsh-1. The overall concordance was 66.8% (48.6% Marsh-1 and 86% Marsh-0) with a Cohen Kappa value of 0.33. In the PCD group, the overall concordance was 63% (45.6% Marsh-1 and 84% Marsh-0) with a Cohen Kappa value of 0.26, while in the CTR group it was 77% (60% Marsh-1, 90% Marsh-0) with a Cohen Kappa value of 0.54. Differences between the two groups were statistically significant (p < 0.05). In conclusion, the concordance of IELs counts between histological and IHC evaluation is low (Kappa Cohen 0.54) in no-CD and even more in PCD patients (0.26). Caution must be paid when classifying a patient as Marsh-0 or Marsh-1 according to the technique used.
上皮内淋巴细胞（IELs）计数是乳糜泻（CD）诊断的核心，可以直接在苏木精和伊红（H&E）染色的石蜡切片上进行，也可以在通过抗CD3免疫组织化学（IHC）染色的最佳切割温度化合物（OCT）包埋的冰冻切片上进行。我们在大量切片样本上评估了这两种技术在马什分级上的一致性。总共纳入了280例肠道结构正常的患者、210例潜在乳糜泻（PCD）患者和70例对照（CTR）。在H&E组织学评估中，136/280被分类为马什-0（显示<25个IELs/100个肠上皮细胞），144例为马什-1，而在IHC评估中，191例被分类为马什-0（显示≤34个CD3+/上皮毫米），89例为马什-1。总体一致性为66.8%（马什-1为48.6%，马什-0为86%），科恩kappa值为0.33。在PCD组中，总体一致性为63%（马什-1为45.6%，马什-0为84%），科恩kappa值为0.26，而在CTR组中为77%（马什-1为60%，马什-0为90%），科恩kappa值为0.54。两组之间的差异具有统计学意义（p<0.05）。总之，在无CD患者中，组织学和IHC评估的IELs计数一致性较低（科恩kappa为0.54），在PCD患者中更低（0.26）。根据所使用的技术将患者分类为马什-0或马什-1时必须谨慎。
