Detection of immunoglobulin light-chain restriction in cutaneous B-cell lymphomas by ultrasensitive bright-field mRNA in situ hybridization
Detection of immunoglobulin light-chain restriction in cutaneous B-cell lymphomas by ultrasensitive bright-field mRNA in situ hybridization
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DOI：
10.1111/cup.12415
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发表时间：
2015-02-01
影响因子：
1.7
通讯作者：
Ma, Xiao-Jun
中科院分区：
文献类型：
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作者：
Minca, Eugen C.;Wang, Hongwei;Ma, Xiao-Jun
背景免疫球蛋白轻链限制检测在b细胞非霍奇金淋巴瘤（B-NHL）的诊断中具有重要意义。流式细胞术，通常用于评估轻链限制，是不切实际的，用于皮肤标本。福尔马林固定石蜡包埋（FFPE）组织的免疫组织化学和常规显色原位杂交（CISH）方法缺乏足够的灵敏度，无法检测未发生浆细胞分化的B-NHL低水平轻链表达。超灵敏亮场mRNA-ISH （BRISH）原位检测皮肤B-NHL已被评估。使用双色BRISH （RNAscope 2xPlex, Advanced Cell Diagnostics）检测27例FFPE皮肤活检和切除患者的DesignKappa/lambda mRNA，这些患者有可用的b细胞PCR克隆研究：16例克隆b细胞病变（6例滤泡中心淋巴瘤，5例边缘区淋巴瘤，3例大b细胞淋巴瘤，2例其他）和11例非克隆b细胞增殖。结果brish在15/16的克隆性b细胞病变和11/11的非克隆性增殖中成功。在15/15个克隆性病变和1/11个非克隆性增生中检测到轻链限制（与克隆性PCR总体一致性为96.1%）。在4/5的边缘区淋巴瘤中，轻链限制在b细胞亚群中检测到强单型mRNA表达，与浆细胞分化一致。结论超灵敏的BRISH可以成功地检测出FFPE皮肤标本中B-NHL的轻链限制，对于CISH或免疫组织化学方法无法解决的病例可能是一种有用的辅助工具。
BackgroundDetection of immunoglobulin light-chain restriction is important in the diagnosis of B-cell non-Hodgkin lymphoma (B-NHL). Flow-cytometry, commonly used to evaluate light-chain restriction, is impractical to be used in cutaneous specimens. Immunohistochemical and conventional chromogenic in situ hybridization (CISH) methods on formalin-fixed-paraffin-embedded (FFPE) tissue lack sufficient sensitivity to detect low-level light-chain expression in B-NHL without plasmacytic differentiation. Ultrasensitive bright-field mRNA-ISH (BRISH) for in situ light-chain detection in cutaneous B-NHL has been assessed.DesignKappa/lambda mRNA was detected using two-color BRISH (RNAscope 2xPlex, Advanced Cell Diagnostics) on 27 FFPE skin biopsies and excisions from patients with available B-cell PCR clonality studies: 16 clonal B-cell lesions (6 follicle center lymphoma, 5 marginal zone lymphoma, 3 large B-cell lymphoma, and 2 other) and 11 non-clonal B-cell proliferations.ResultsBRISH was successful in 15/16 clonal B-cell lesions and 11/11 non-clonal proliferations. Light-chain restriction was detected in 15/15 clonal lesions and in 1/11 non-clonal proliferations (96.1% overall concordance with clonality PCR). In 4/5 marginal zone lymphomas, light-chain restriction was detected as strong monotypic mRNA expression in a B-cell subset, consistent with plasmacytic differentiation.ConclusionUltrasensitive BRISH can successfully detect light-chain restriction in B-NHL from FFPE skin specimens and may be a useful adjunct ancillary tool in cases not resolved by CISH or immunohistochemical methods.