Clinical and Experimental Nephrology ( IF 2.2 ) Pub Date : 2024-03-04 , DOI: 10.1007/s10157-024-02464-z Shintaro Komatsu 1, 2, 3 , Noritoshi Kato 1 , Hiroki Kitai 1, 3 , Yoshio Funahashi 1 , Yuhei Noda 1, 3 , Shoma Tsubota 3 , Akihito Tanaka 1 , Yuka Sato 1 , Kayaho Maeda 1 , Shoji Saito 1 , Kazuhiro Furuhashi 1 , Takuji Ishimoto 1 , Tomoki Kosugi 1 , Shoichi Maruyama 1 , Kenji Kadomatsu 3
|
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.
中文翻译：
检测和探索血浆中肾源性细胞外囊泡
背景
细胞外囊泡（EV）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 EV，它们主要来源于泌尿道细胞。然而，血浆中肾源性EV的检测和应用仍不确定。
方法
我们使用来自健康对照和肾移植患者的临床样本检查了血浆中的肾源性小 EV（sEV），这些小 EV 据称是从肾系膜和肾小球内皮细胞释放的。来自健康对照的血浆经过超速离心，然后进行珠上流式细胞术，针对α8整合素（一种对系膜细胞具有特异性的抗原）。为了确认外周血中是否存在肾源性 sEV，对 ABO 不相容肾移植受者的血浆进行超速离心，然后对供体血型抗原进行蛋白质印迹分析。
结果
免疫组织化学和免疫电子显微镜证实肾系膜细胞及其 sEV 中存在 α8 整合素表达。使用珠上流式细胞术成功检测到 CD9-α8 整合素双阳性 sEV。蛋白质印迹分析进一步显示，在接受 B 型血供体肾脏的非 B 型受者中，移植的肾源性 sEV 含有 B 型抗原。值得注意的是，经历移植肾丧失的患者表现出含有供体血型抗原的 sEV 信号减弱。
结论
我们的研究结果证明了血浆中肾源性 sEV 在未来肾脏疾病研究中的潜在用途。
京公网安备 11010802027423号