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Immunophenotyping without antibodies. New perspectives for lymphoma characterization

机译：没有抗体的免疫分型。淋巴瘤表征的新观点

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AIMS: Accurate classification of haematological malignancies is a prerequisite for their correct diagnosis, prognosis and therapy. Clear classification of lymphomas is often hindered by the limited number of available cell surface protein markers that are suitable for immunophenotyping. A systematic and quantitative analysis of cell surface proteins is thus required to identify new protein markers on lymphoma subtypes in an unbiased and discovery-driven approach. METHODS: Nine Hodgkin and non-Hodgkin B cell lines of diffuse large cell type and mediastinal type were investigated by cell surface capture (CSC) technology, a mass spectrometry-based method to identify cell surface glycoproteins. Selected proteins are verified by antibody-based methods, including flow cytometry and immunohistochemistry on cell line arrays. RESULTS: A total of 747 predicted transmembrane proteins were identified from all cell lines, including 142 CD (cluster of differentiation) annotated proteins. A group of differentially expressed cell surface glycoproteins between Hodgkin and non-Hodgkin B cell lines was revealed via quantitative CSC technology. In addition to classical and expected CD molecules such as CD20 and CD30, less frequently expressed molecules such as CD2 on Hodgkin lymphoma (HL) cell lines were identified by CSC and verified by immunohistochemistry in cell lines and primary lymphoma tissue. A panel of CSC-identified differentiation glycoprotein candidates is currently under investigation on tissue microarrays (TMAs) from patient samples.
机译：目的：正确分类血液恶性肿瘤是正确诊断，预后和治疗的前提。淋巴瘤的清晰分类通常会受到数量有限的适用于免疫表型的可用细胞表面蛋白标志物的阻碍。因此，需要以无偏见和发现驱动的方法对细胞表面蛋白进行系统和定量的分析，以鉴定淋巴瘤亚型上的新蛋白标记。方法：利用细胞表面捕获（CSC）技术研究了九种弥散性大细胞类型和纵隔类型的霍奇金和非霍奇金B细胞系，这是一种基于质谱的方法来鉴定细胞表面糖蛋白。通过基于抗体的方法（包括流式细胞仪和细胞系阵列上的免疫组织化学）验证所选蛋白。结果：从所有细胞系中共鉴定出747个预测跨膜蛋白，包括142个CD（分化簇）注释蛋白。通过定量CSC技术揭示了一组霍奇金和非霍奇金B细胞系之间差异表达的细胞表面糖蛋白。除了经典的和预期的CD分子（例如CD20和CD30）以外，霍奇金淋巴瘤（HL）细胞株上表达较不频繁的分子（例如CD2）也通过CSC进行了鉴定，并通过免疫组织化学在细胞株和原发性淋巴瘤组织中进行了验证。目前正在对来自患者样品的组织微阵列（TMA）进行一组CSC鉴定的分化糖蛋白候选物的研究。

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