Comparison of Different Antibody Clones for Immunohistochemistry Detection of Programmed Cell Death Ligand 1 (PD-L1) on Non-Small Cell Lung Carcinoma
Comparison of Different Antibody Clones for Immunohistochemistry Detection of Programmed Cell Death Ligand 1 (PD-L1) on Non-Small Cell Lung Carcinoma
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DOI：
10.1097/pai.0000000000000531
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发表时间：
2018-02-01
影响因子：
1.6
通讯作者：
Rodriguez-Canales, Jaime
中科院分区：
文献类型：
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作者：
Parra, Edwin R.;Villalobos, Pamela;Rodriguez-Canales, Jaime
程序性细胞死亡配体 1 (PD-L1) 是一种主要的免疫检查点蛋白，可介导抗肿瘤免疫抑制和反应。初步数据表明，在福尔马林固定和石蜡包埋的组织中使用免疫组织化学 (IHC) 进行检测可以预测对 PD-1/PD-L1 治疗的临床反应。在诊断病理学中，必须使用经过验证的 IHC 进行计数，以可靠地检测 PD-L1 阳性病例。本研究旨在比较和验证不同的 PD-L1 商业克隆，并确定哪些克隆可以被外科病理学家可靠地用于检测人类癌症组织中的 PD-L1 表达。对八个市售 PD-L1 克隆进行了测试，并与非市售 PD-L1 抗体克隆 5H1 进行了比较。使用细胞系和人体组织进行蛋白质印迹和 IHC 来验证这些克隆。在所有 PD-L1 抗体中，只有克隆 E1L3N、E1J2J、SP142、28-8、22C3 和 SP263 通过了蛋白质印迹和 IHC 验证，提供了与克隆 5H1 相似的模式，然后在置于 9 个组织微阵列中的 259 个非小细胞肺癌病例中进行了测试。在所有案例中，仅包含 2 核案例（185 例）。对于所有选定的抗体，在肿瘤区室和基质区室中的中位 PD-L1 H 评分之间发现显着正相关。总体而言，185例中的56例被所有克隆检测为表达膜PD-L1的恶性细胞阳性病例。然而，与其他克隆相比，克隆 SP263 识别出更多的 PD-L1 阳性病例。我们的结果表明，克隆 E1L3N、E1J2J、SP142、28-8、22C3 和 SP263 提供与克隆 5H1 相当的阳性膜染色模式。这些商业克隆具有可比性，但需要病理学家进行仔细评估，以尽量减少积极误解的错误。
Programmed cell death ligand 1 (PD-L1) is a major immune checkpoint protein that mediates antitumor immune suppression and response. Preliminary data suggest that its detection using immunohistochemistry (IHC) in formalin-fixed and paraffin-embedded tissues may predict clinical response to PD-1/PD-L1 therapy. In diagnostic pathology, it is essential to count with a validated IHC that can reliably detect PD-L1-positive cases. The present study was conducted to compare and validate different PD-L1 commercial clones and identify which ones can be reliably used by surgical pathologist to detect PD-L1 expression in human cancer tissues. Eight commercial available PD-L1 clones were tested and compared with a noncommercial PD-L1 antibody clone 5H1. Western blot and IHC using cell lines and human tissues were used to validate these clones. From all PD-L1 antibodies, only the clones E1L3N, E1J2J, SP142, 28-8, 22C3, and SP263 passed the Western blot and IHC validation, providing similar pattern than the clone 5H1 and then they were tested in 259 non-small cell lung cancer cases placed in 9 tissue microarrays. Among all cases, only those with 2 cores were included (185 cases). Positive and significant correlation was found between the median PD-L1 H-score in tumor and stroma compartments, for all selected antibodies. Overall, 56 of 185 cases were detected as positive cases in malignant cells expressing membranous PD-L1 by all the clones. However, the clone SP263 identified more PD-L1-positive cases compared with the other clones. Our results show that clones E1L3N, E1J2J, SP142, 28-8, 22C3, and SP263 provide positive membrane staining pattern comparable with clone 5H1. These commercial clones are comparable, but a careful evaluation by the pathologist is necessary to minimize error of positive misinterpretations.