急性髓细胞性白血病(AML)患者血浆中积累的小细胞外囊泡(sEV)是白血病和非恶性细胞产生的囊泡混合物。源自白血病母细胞的sEV可以作为AML对治疗反应的潜在非侵入性生物标志物。从患者血浆中分离爆炸来源的sEV,我们使用对白血病相关抗原(LAA)特异的单克隆抗体(mAb)和对四跨膜蛋白混合物(CD9,CD63和CD81)特异的mAb,开发了一种基于生物打印微阵列的免疫测定法.我们确定了LAA+sEV相对于总血浆sEV的比例(LAA+/总sEV比率),during,化疗后。在AML诊断时,患者的LAA+/总sEV比率显著高于健康供者(HDs).在诱导化疗后达到完全缓解(CR)的患者中,LAA+/总sEV比率在每个化疗周期后显著降低至HDs中的水平.相比之下,治疗后患有持续性白血病的AML患者的LAA+/总sEV比率在治疗期间和之后仍然升高,这些患者骨髓中白血病母细胞的百分比也是如此。LAA+/总sEV比率作为白血病对治疗反应的有希望的非侵入性生物标志物出现。
The small extracellular vesicles (sEV) accumulating in acute myeloid leukemia (AML) patients\' plasma are mixtures of vesicles produced by leukemic and non-malignant cells. sEV originating from leukemia blasts could serve as potential non-invasive biomarkers of AML response to therapy. To isolate blast-derived sEV from patients\' plasma, we developed a bioprinted microarray-based immunoassay using monoclonal antibodies (mAbs) specific for leukemia-associated antigens (LAAs) and mAbs specific for a mix of tetraspanins (CD9, CD63, and CD81). We determined the proportion of LAA+ sEV relative to total plasma sEV (the LAA+/total sEV ratio) in serially collected samples of newly diagnosed AML patients prior to, during, and after chemotherapy. At AML diagnosis, the LAA+/total sEV ratio was significantly higher in patients than in healthy donors (HDs). In patients who achieved complete remission (CR) after induction chemotherapy, the LAA+/total sEV ratios significantly decreased after each chemotherapy cycle to levels seen in HDs. In contrast, the LAA+/total sEV ratios in AML patients with persistent leukemia after therapy remained elevated during and after therapy, as did the percentage of leukemic blasts in these patients\' bone marrows. The LAA+/total sEV ratio emerges as a promising non-invasive biomarker of leukemia response to therapy.