{"title":"Detecting and exploring kidney-derived extracellular vesicles in plasma.","authors":"Shintaro Komatsu, Noritoshi Kato, Hiroki Kitai, Yoshio Funahashi, Yuhei Noda, Shoma Tsubota, Akihito Tanaka, Yuka Sato, Kayaho Maeda, Shoji Saito, Kazuhiro Furuhashi, Takuji Ishimoto, Tomoki Kosugi, Shoichi Maruyama, Kenji Kadomatsu","doi":"10.1007/s10157-024-02464-z","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Extracellular vesicles (EVs) have received considerable attention as ideal biomarkers for kidney diseases. Most reports have focused on urinary EVs, that are mainly derived from the cells in the urinary tract. However, the detection and the application of kidney-derived EVs in plasma remains uncertain.</p><p><strong>Methods: </strong>We examined the kidney-derived small EVs (sEVs) in plasma that were supposedly released from renal mesangial and glomerular endothelial cells, using clinical samples from healthy controls and patients with kidney transplants. Plasma from healthy controls underwent ultracentrifugation, followed by on-bead flow cytometry, targeting α8 integrin, an antigen-specific to mesangial cells. To confirm the presence of kidney-derived sEVs in peripheral blood, plasma from ABO-incompatible kidney transplant recipients was ultracentrifuged, followed by western blotting for donor blood type antigens.</p><p><strong>Results: </strong>Immunohistochemistry and immunoelectron microscopy confirmed α8 integrin expression in kidney mesangial cells and their sEVs. The CD9-α8 integrin double-positive sEVs were successfully detected using on-bead flow cytometry. Western blot analysis further revealed transplanted kidney-derived sEVs containing blood type B antigens in non-blood type B recipients, who had received kidneys from blood type B donors. Notably, a patient experiencing graft kidney loss exhibited diminished signals of sEVs containing donor blood type antigens.</p><p><strong>Conclusion: </strong>Our findings demonstrate the potential usefulness of kidney-derived sEVs in plasma in future research for kidney diseases.</p>","PeriodicalId":10349,"journal":{"name":"Clinical and Experimental Nephrology","volume":" ","pages":"617-628"},"PeriodicalIF":2.2000,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11190017/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Clinical and Experimental Nephrology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s10157-024-02464-z","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/3/4 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"UROLOGY & NEPHROLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Background: Extracellular vesicles (EVs) have received considerable attention as ideal biomarkers for kidney diseases. Most reports have focused on urinary EVs, that are mainly derived from the cells in the urinary tract. However, the detection and the application of kidney-derived EVs in plasma remains uncertain.
Methods: We examined the kidney-derived small EVs (sEVs) in plasma that were supposedly released from renal mesangial and glomerular endothelial cells, using clinical samples from healthy controls and patients with kidney transplants. Plasma from healthy controls underwent ultracentrifugation, followed by on-bead flow cytometry, targeting α8 integrin, an antigen-specific to mesangial cells. To confirm the presence of kidney-derived sEVs in peripheral blood, plasma from ABO-incompatible kidney transplant recipients was ultracentrifuged, followed by western blotting for donor blood type antigens.
Results: Immunohistochemistry and immunoelectron microscopy confirmed α8 integrin expression in kidney mesangial cells and their sEVs. The CD9-α8 integrin double-positive sEVs were successfully detected using on-bead flow cytometry. Western blot analysis further revealed transplanted kidney-derived sEVs containing blood type B antigens in non-blood type B recipients, who had received kidneys from blood type B donors. Notably, a patient experiencing graft kidney loss exhibited diminished signals of sEVs containing donor blood type antigens.
Conclusion: Our findings demonstrate the potential usefulness of kidney-derived sEVs in plasma in future research for kidney diseases.
背景：细胞外囊泡（EVs）作为肾脏疾病的理想生物标志物受到了广泛关注。大多数报告都集中在尿液 EVs 上，这些 EVs 主要来源于尿路细胞。然而，血浆中肾脏衍生EVs的检测和应用仍不确定：方法：我们利用健康对照组和肾移植患者的临床样本研究了血浆中肾脏衍生的小EVs（sEVs），这些EVs可能是从肾间质和肾小球内皮细胞中释放出来的。健康对照组的血浆经过超速离心处理，然后进行珠上流式细胞仪检测，检测对象是系膜细胞特异性抗原α8整合素。为了证实外周血中存在肾源性 sEV，对 ABO 不相容肾移植受者的血浆进行了超速离心，然后对供体血型抗原进行了 Western 印迹检测：免疫组织化学和免疫电镜检查证实肾间质细胞及其 sEV 中表达了 α8 整合素。使用珠上流式细胞术成功检测到了 CD9-α8 整合素双阳性的 sEV。Western 印迹分析进一步发现，在接受 B 型血供体肾脏的非 B 型血受体中，移植肾衍生的 sEV 含有 B 型血抗原。值得注意的是，一名移植肾缺失的患者体内含有供体血型抗原的 sEVs 信号减弱：我们的研究结果表明，血浆中肾脏来源的 sEVs 在未来肾脏疾病研究中具有潜在用途。
期刊介绍：
Clinical and Experimental Nephrology is a peer-reviewed monthly journal, officially published by the Japanese Society of Nephrology (JSN) to provide an international forum for the discussion of research and issues relating to the study of nephrology. Out of respect for the founders of the JSN, the title of this journal uses the term “nephrology,” a word created and brought into use with the establishment of the JSN (Japanese Journal of Nephrology, Vol. 2, No. 1, 1960). The journal publishes articles on all aspects of nephrology, including basic, experimental, and clinical research, so as to share the latest research findings and ideas not only with members of the JSN, but with all researchers who wish to contribute to a better understanding of recent advances in nephrology. The journal is unique in that it introduces to an international readership original reports from Japan and also the clinical standards discussed and agreed by JSN.
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