程序性细胞死亡1-配体1在不同免疫组织化学染色方法的一致性比较

doi:10.19723/j.issn.1671-167X.2023.02.020

摘要/Abstract

摘要：

目的: 比较程序性细胞死亡1-配体1(programmed cell death 1-ligand 1, PD-L1，克隆号E1L3N、22C3、SP263)在不同免疫组织化学染色方法的一致性。方法: 第一步，筛选最优流程：采用PD-L1(克隆号E1L3N)抗体分别按推荐流程、自建流程①、自建流程②与自建流程③，对5例扁桃体组织完成平行免疫组织化学染色，专科病理医生对全部切片进行质量评分(0~6分)，筛选出评分最高的流程；第二步，用最优流程与两种标准流程的评分结果进行一致性比较：选取近两年北京大学第一医院病理最终诊断为非小细胞肺癌的标本共32例，用筛选出的自建流程①、与SP263标准流程及22C3标准流程对32病例各自进行平行染色，全部染色片由专科病理医师行阳性肿瘤细胞比例评分(tumor proportion score，TPS)。评分结果按＜1%，≥1%至＜10%，≥10%至＜50%，≥50%分组，分析PD-L1检测抗体克隆号E1L3N与22C3、SP263染色结果的一致性。结果: 扁桃体染色切片质量评分(0~6分)如下：推荐流程为5、5、5、5、5分；自建流程①为5、6、6、5、6分；自建流程②为4、4、4、4、4分；自建流程③为3、3、3、3、3分。自建流程①的结果总体评分最高，选用自建流程①完成32例非小细胞肺癌病例的免疫组组织化学染色，TPS评分为：自建流程①，＜1%共6例，≥1%至＜10%共5例，≥10%至 < 50%共10例，≥50%共11例；22C3标准流程，＜1%共5例，≥1%至＜10%共3例，≥10%至＜50%共13例，≥50%共11例；SP263标准流程，＜1%共7例，≥1%至＜10%共4例，≥10%至＜50%共11例，≥50%共10例。一致性检验结果为：自建流程①和22C3标准流程，κ值为0.736(P ＜0.001)，一致性好；自建流程①和SP263标准流程，κ值为0.914(P ＜0.001)，一致性极好。结论: 采用PD-L1(克隆号E1L3N)抗体，在罗氏Ventana Benchmark GX平台，通过自建染色流程①完成，可获得良好质量的染色切片；在非小细胞肺癌病例的检测中，自建染色流程①与22C3标准流程和SP263标准流程一致性好。

关键词: 非小细胞肺癌, 程序性细胞死亡1-配体1, 克隆号E1L3N, 免疫组织化学染色

Abstract:

Objective: To compare the consistency of programmed cell death 1-ligand 1 (PD-L1, clone E1L3N, 22C3, SP263) in different immunohistochemical staining methods. 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. Results: 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 κ value for self-built process ① and 22C3 standard procedure was 0.736 (P < 0.001), the agreement was good; the κ value for self-built process ① and SP263 standard procedure was 0.914 (P < 0.001), the agreement was very good. Conclusion: 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.

Key words: Non small cell lung carcinoma, Programmed cell death 1-ligand 1, Colne E1L3N, Immunohistochemistr

中图分类号:

R365

李东,邸吉廷,熊焰. 程序性细胞死亡1-配体1在不同免疫组织化学染色方法的一致性比较[J]. 北京大学学报（医学版）, 2023, 55(2): 339-342.

Dong LI,Ji-ting DI,Yan XIONG. Consistency comparison of programmed cell death 1-ligand 1 in different immuno-histochemical staining methods[J]. Journal of Peking University (Health Sciences), 2023, 55(2): 339-342.

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参考文献 11

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