Li D, DI J T, Xiong Y
Department of Pathology, Peking University First Hospital, Beijing 100034, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2023 Apr 18;55(2):339-342. doi: 10.19723/j.issn.1671-167X.2023.02.020.
To compare the consistency of programmed cell death 1-ligand 1 (PD-L1, clone E1L3N, 22C3, SP263) in different immunohistochemical staining methods.
The first step was to select the optimal process: The PD-L1(clone E1L3N) antibody recommended process, self-built process ①, self-built process ② and self-built process ③ were used to perform immunohistochemical staining in 5 cases of tonsil tissue. The quality of all slides was scored by expert pathologists (0-6 points). The process with the highest score was selected. The second step was to compare the consistency between the optimal procedure and the two standard procedures. Thirty-two cases of lung non-small cell carcinoma diagnosed by pathology in Peking University First Hospital in the past two years were randomly selected. The 32 cases were stained in parallel with the SP263 and 22C3 standard procedures, and all stained slides were scored by specialized pathologists for tumor proportion score (TPS). The scoring results were grouped according to < 1%, ≥1% to < 10%, ≥10% to < 50%, and ≥50%. The consistency of PD-L1 detection antibody clone E1L3N and 22C3, E1L3N and SP263 staining results was analyzed.
Tonsil stained slides scores (0-6 points) were as follows: The recommended protocol was 5, 5, 5, 5 and 5. The self-built process ① was 5, 6, 6, 5 and 6. The self-built process ② was 4, 4, 4, 4 and 4.The self-built process ③ was 3, 3, 3, 3 and 3. The self-built process ① was the best with the highest score. The TPSs of 32 non small cell lung carcinoma (NSCLC) cases were as follows: Of self-built process ①, 6 cases were lower than 1%, 5 cases were from 1% to 10%, 10 cases were from 10% to 50%, and 11 cases were higher than 50%; of 22C3 standard procedure, 5 cases were lower than 1%, 3 cases were from 1% to 10%, 13 cases were from 10% to 50%, 11 cases were higher than 50%; of SP263 standard procedure, 7 cases were lower than 1%, 4 cases were from 1% to 10%, 11 cases were from 10% to 50%, 10 cases were higher than 50%. The results of the consistency test were as follows: The value for self-built process ① and 22C3 standard procedure was 0.736 ( < 0.001), the agreement was good; the value for self-built process ① and SP263 standard procedure was 0.914 ( < 0.001), the agreement was very good.
The immunostaining using PD-L1(E1L3N) with validated self-built staining protocol ① by Ventana Benchmark GX platform can obtain high quality of slides, and the TPSs based on these slides are in good agreement with 22C3 and SP263 standard procedures.
比较程序性死亡蛋白1配体1（PD-L1，克隆号E1L3N、22C3、SP263）在不同免疫组织化学染色方法中的一致性。
第一步是选择最佳流程：采用PD-L1（克隆号E1L3N）抗体推荐流程、自建流程①、自建流程②和自建流程③对5例扁桃体组织进行免疫组织化学染色。由病理专家对所有切片质量进行评分（0 - 6分）。选择得分最高的流程。第二步是比较最佳流程与两种标准流程之间的一致性。随机选取北京大学第一医院过去两年经病理诊断的32例肺非小细胞癌病例。这32例病例分别采用SP263和22C3标准流程进行平行染色，所有染色切片由专业病理学家进行肿瘤比例评分（TPS）。评分结果按<1%、≥1%至<10%、≥10%至<50%、≥50%进行分组。分析PD-L1检测抗体克隆号E1L3N与22C3、E1L3N与SP263染色结果的一致性。
扁桃体染色切片评分（0 - 6分）如下：推荐流程的评分为5、5、5、5和5分。自建流程①为5、6、6、5和6分。自建流程②为4、4、4、4和4分。自建流程③为3、3、3、3和3分。自建流程①得分最高，效果最佳。32例非小细胞肺癌（NSCLC）病例的TPS如下：自建流程①中，6例低于1%，5例为1%至10%，10例为10%至50%，11例高于50%；22C3标准流程中，5例低于1%，3例为1%至10%，13例为10%至50%，11例高于50%；SP263标准流程中，7例低于1%，4例为1%至10%，11例为10%至50%，10例高于50%。一致性检验结果如下：自建流程①与22C3标准流程的κ值为0.736（P<0.001），一致性良好；自建流程①与SP263标准流程的κ值为0.914（P<0.001），一致性非常好。
采用Ventana Benchmark GX平台经验证的自建染色流程①对PD-L1（E1L3N）进行免疫染色可获得高质量切片，基于这些切片的TPS与22C3和SP263标准流程具有良好的一致性。
