PDF(Pubmed)
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.
CONCLUSIONS: Our findings demonstrate the potential usefulness of kidney-derived sEVs in plasma in future research for kidney diseases.
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.
CONCLUSIONS: Our findings demonstrate the potential usefulness of kidney-derived sEVs in plasma in future research for kidney diseases.
摘要：
背景：细胞外囊泡（EV）作为肾脏疾病的理想生物标志物已受到广泛关注。大多数报告都集中在泌尿系统上，主要来自泌尿道中的细胞。然而,肾源性EV在血浆中的检测和应用仍不确定.
方法：我们检查了从肾系膜和肾小球内皮细胞释放的血浆中的肾源性小EV（sEV），使用健康对照和肾移植患者的临床样本。来自健康对照的血浆进行超速离心，然后是珠子上的流式细胞术，靶向α8整合素，肾小球膜细胞特异性抗原.为了确认外周血中存在肾源性sEV，来自ABO不相容的肾移植受者的血浆被超速离心，然后是供体血型抗原的免疫印迹。
结果：免疫组织化学和免疫电子显微镜证实了肾系膜细胞及其sEV中α8整合素的表达。使用珠上流式细胞术成功检测了CD9-α8整合素双阳性sEV。蛋白质印迹分析进一步显示，非血型B型受体中含有血型B型抗原的移植肾源性sEV，从B型血捐献者那里获得肾脏。值得注意的是,一名移植肾丢失的患者表现出含有供体血型抗原的sEV信号减弱.
结论：我们的发现证明了肾源性sEV在未来肾脏疾病研究中的潜在用途。
方法：我们检查了从肾系膜和肾小球内皮细胞释放的血浆中的肾源性小EV（sEV），使用健康对照和肾移植患者的临床样本。来自健康对照的血浆进行超速离心，然后是珠子上的流式细胞术，靶向α8整合素，肾小球膜细胞特异性抗原.为了确认外周血中存在肾源性sEV，来自ABO不相容的肾移植受者的血浆被超速离心，然后是供体血型抗原的免疫印迹。
结果：免疫组织化学和免疫电子显微镜证实了肾系膜细胞及其sEV中α8整合素的表达。使用珠上流式细胞术成功检测了CD9-α8整合素双阳性sEV。蛋白质印迹分析进一步显示，非血型B型受体中含有血型B型抗原的移植肾源性sEV，从B型血捐献者那里获得肾脏。值得注意的是,一名移植肾丢失的患者表现出含有供体血型抗原的sEV信号减弱.
结论：我们的发现证明了肾源性sEV在未来肾脏疾病研究中的潜在用途。
