{"title":"Evaluation of Constitutive or Induced Activation State of the VLA-4 Integrin in Human and Murine Samples.","authors":"Erika Tissino, Antonella Zucchetto, Federico Pozzo, Tanja Nicole Hartmann, Valter Gattei","doi":"10.1007/978-1-0716-4442-3_4","DOIUrl":null,"url":null,"abstract":"<p><p>The integrin heterodimer CD49d/CD29 (a.k.a. Very Late Antigen-4, VLA-4) mediates cell-cell and cell-matrix interaction through binding to its specific ligands. VLA-4 can be present on the cell surface at different conformation states that affect the binding affinity for the ligands. In chronic lymphocytic leukemia (CLL), higher VLA-4 levels have been demonstrated to be associated with a worse prognosis both in the chemo-immunotherapy era and in the BCR inhibitor setting, in keeping with the role of VLA-4 as a key molecule favoring CLL cell localization in protective niches of bone marrow and lymph nodes. Here, we describe functional flow cytometry-based methods to assess the activation state of the VLA-4 integrin, applicable in both human and murine settings, as well as in fresh or thawed samples. A specific \"R\" script for analyzing flow cytometry data is also provided.</p>","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2909 ","pages":"45-60"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in molecular biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/978-1-0716-4442-3_4","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0
Abstract
The integrin heterodimer CD49d/CD29 (a.k.a. Very Late Antigen-4, VLA-4) mediates cell-cell and cell-matrix interaction through binding to its specific ligands. VLA-4 can be present on the cell surface at different conformation states that affect the binding affinity for the ligands. In chronic lymphocytic leukemia (CLL), higher VLA-4 levels have been demonstrated to be associated with a worse prognosis both in the chemo-immunotherapy era and in the BCR inhibitor setting, in keeping with the role of VLA-4 as a key molecule favoring CLL cell localization in protective niches of bone marrow and lymph nodes. Here, we describe functional flow cytometry-based methods to assess the activation state of the VLA-4 integrin, applicable in both human and murine settings, as well as in fresh or thawed samples. A specific "R" script for analyzing flow cytometry data is also provided.
整合素异源二聚体CD49d/CD29（又名Very Late Antigen-4， vla4）通过与其特异性配体的结合介导细胞-细胞和细胞-基质的相互作用。vla4可以以不同的构象状态存在于细胞表面，从而影响对配体的结合亲和力。在慢性淋巴细胞白血病（CLL）中，较高的vla4水平已被证明与化疗免疫治疗时代和BCR抑制剂环境下较差的预后相关，这与vla4作为CLL细胞在骨髓和淋巴结保护龛中定位的关键分子的作用保持一致。在这里，我们描述了基于功能流式细胞术的方法来评估VLA-4整合素的激活状态，适用于人类和小鼠环境，以及新鲜或解冻的样品。还提供了用于分析流式细胞术数据的特定“R”脚本。
期刊介绍：
For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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