摘要
目的探讨通过改良的免疫组织化学方法,检测唾液酸化CD15(CD15s)在经典霍奇金淋巴瘤肿瘤细胞中的表达情况。方法从北京大学肿瘤医院病理科2009—2024年收集的53例经典霍奇金淋巴瘤病例中,筛选出21例CD15表达为阴性或仅少数细胞呈弱阳性的样本。使用Leica全自动免疫组织化学染色机,分别采用3种不同克隆号(MMA、Carb3和IHC527)的CD15抗体进行染色。应用不同浓度与孵育时间的唾液酸酶对样本进行消化处理,再以克隆号MMA的CD15抗体进行免疫组织化学染色。同时,采用多重免疫荧光技术对CD15(MMA)与CD30(JCM182)进行共同标记染色,并利用APTIME和HALO软件进行染色结果的对比分析。结果患者男性30例,女性23例,年龄范围14~73岁,中位年龄32(26,46)岁。在14例CD15阴性及7例仅少数细胞弱阳性病例中,3种不同克隆号的CD15抗体均未能提高CD15的阳性检出率,且三者间差异无统计学意义(P>0.05)。经唾液酸酶消化后,组织形态保持完好;其中,在37℃条件下使用1∶1稀释的唾液酸酶孵育1 h,可获得最佳的CD15s染色效果。经此处理,Hodgkin Reed-Sternberg细胞(HRS细胞)中CD15s阳性细胞比例显著提升(P<0.001)。结论唾液酸酶消化处理可有效显示经典霍奇金淋巴瘤中CD15s的表达,从而提高其在HRS细胞中的检出率。
Objective To detect the expression of sialylated CD15(CD15s)in the tumor cells of classical Hodgkin lymphoma using a modified immunohistochemical approach.Methods From 2009 to 2024,53 cases of classical Hodgkin lymphoma were collected in the Department of Pathology,Peking University Cancer Hospital,in which 21 cases that were CD15-negative or showed only focal weak positivity were selected.Immunohistochemical staining for CD15 was performed on a Leica automated stainer using three different antibody clones(MMA,Carb3,and IHC527).Tissue sections were digested with sialidase at varying concentrations and incubation times,followed by immunohistochemical staining with the MMA clone.Multiplex immunofluorescence was applied for co-staining of CD15(MMA)and CD30(JCM182),and analysis was conducted using APTIME and HALO software.Results There were 30 male patients and 23 female patients,with an age range of 14 to 73 years and a median age of 32(26,46)years.None of the three CD15 antibody clones significantly improved the CD15 positive rate in the 14 completely negative and 7 weakly positive cases,with no notable differences observed among the clones(P>0.05).After sialidase digestion,tissue morphology remained well-preserved.Optimal CD15 staining was achieved with a 1∶1 diluted sialidase incubated at 37℃for one hour.This treatment significantly enhanced the detection rate of CD15 antigen in Hodgkin Reed-Sternberg cells(P<0.001).Conclusion Sialidase digestion effectively unveils sialylated CD15 expression in classical Hodgkin lymphoma,markedly improving its detection in HRS cells.
作者
马秀利
周立新
时云飞
赖玉梅
刁新婷
王宏伟
李向红
Ma Xiuli;Zhou Lixin;Shi Yunfei;Lai Yumei;Diao Xinting;Wang Hongwei;Li Xianghong(Department of Pathology,Peking University Cancer Hospital/Beijing Cancer Institute,Key Laboratory of Carcinogenesis and Translational Research Ministry of Education,Beijing 100142,China;Department of Pathology,Fourth Medical Center of People's Liberation Army General Hospital,Beijing 100037,China)
出处
《中华病理学杂志》
北大核心
2025年第12期1282-1287,共6页
Chinese Journal of Pathology
基金
血液肿瘤真实世界专项研究基金(KY-202409-0)。
