Analysis of Galectin-Binding Receptors on B Cells.
Analysis of Galectin-Binding Receptors on B Cells.
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DOI：
10.1007/978-1-0716-2055-7_30
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发表时间：
2022
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β-半乳糖苷结合凝集素家族（称为半乳糖凝集素）在疾病发展中的作用已经以惊人的速度发展。半乳糖凝集素及其聚糖反受体配体现在被认为是恶性和转移性进展、肿瘤免疫逃避、自身免疫和免疫稳态的关键功能决定因素。它们在这些过程中的影响是通过在肿瘤、免疫和基质细胞区室中的协调表达引起的。虽然在相关研究工作中半乳糖凝集素水平的分析通常通过免疫测定和RT-qPCR进行，但细胞表面上天然形式的聚糖反受体配体的检测和鉴定已经滞后。在这份报告中，我们提出了检测和鉴定半乳糖凝集素（Gal）-3和Gal-9的聚糖反受体配体的方法，这两种半乳糖凝集素是癌症和免疫学研究的十字准线。作为一个模型，我们将描述（1）从新鲜扁桃体组织中分离人B细胞亚群，（2）测定人B细胞上的Gal-3/−9结合活性，以及（3）鉴定人B细胞表面的Gal-3/−9配体。当然，这些方法可以在任何细胞类型上实施，以提供能够传递半乳糖凝集素介导的表型的细胞和分子环境。在特异性反受体配体上建立半乳糖凝集素结合活性可以帮助发掘能够传递疾病进展所需的细胞信号的潜在关键决定因素。这些进展开辟了新的研究途径，导致新的治疗靶点和方法。
The reported roles of the β-galactoside-binding lectin family, known as galectins, in disease development have been advancing at a remarkable pace. Galectins and their glycan counter-receptor ligands are now considered key functional determinants in malignant and metastatic progression, tumor immune evasion, autoimmunity, and immune homeostasis. Their influence in these processes is elicited through coordinated expression in tumor, immune and stromal cellular compartments. While analysis of galectin levels in related research efforts is routinely performed through immunoassays and RT-qPCR, detection, and identification of glycan counter-receptor ligands in their native form on the cell surface has lagged. In this report, we present methods to detect and identify glycan counter-receptor ligands to galectin (Gal)-3 and Gal-9—two galectins at the crosshairs of cancer and immunology research. As a model, we will describe (1) isolation of human B-cell subsets from fresh tonsil tissue, (2) assaying of Gal-3/−9-binding activities on human B cells, and (3) identifying Gal-3/−9 ligands on human B-cell surfaces. These methods, of course, can be implemented on any cell type to provide a cellular and molecular context capable of transmitting a galectin-mediated phenotype. Establishing a galectin-binding activity on specific counter-receptor ligand(s) can help unearth potential critical determinants capable of delivering cellular signals required for disease progression. These advances open new avenues of research investigation that result in novel therapeutic targets and approaches.