来自骨髓间充质基质细胞的细胞外囊泡可支持人抗体分泌细胞的存活。
Extracellular vesicles from bone marrow-derived mesenchymal stromal cells support survival of human antibody secreting cells.
作者信息
Nguyen Doan C, Lewis Holly C, Joyner Chester, Warren Vivien, Xiao Haopeng, Kissick Haydn T, Wu Ronghu, Galipeau Jacques, Lee F Eun-Hyung
机构信息
Division of Pulmonary Allergy, Critical Care, & Sleep Medicine, Emory University, Atlanta, GA, USA.
Departments of Pediatrics and Hematology & Medical Oncology, Emory University School of Medicine, Atlanta, GA, USA.
出版信息
J Extracell Vesicles. 2018 Apr 26;7(1):1463778. doi: 10.1080/20013078.2018.1463778. eCollection 2018.
Extracellular vesicles (EVs) from bone marrow (BM)-derived mesenchymal stromal cells (BM-MSC) are novel mechanisms of cell-cell communication over short and long distances. BM-MSC have been shown to support human antibody secreting cells (ASC) survival , but whether the crosstalk between the MSC-ASC interaction can occur via EVs is not known. Thus, we evaluated the role of EVs in ASC survival and IgG secretion. EVs were isolated from irradiated and non-irradiated primary BM-MSC and were quantified. They were further characterized by electron microscopy (EM) and CD63 and CD81 immuno-gold EM staining. Human ASC were isolated via fluorescence-activated cell sorting (FACS) and cultured with the EV fractions, the EV-reduced fractions, or conventional media. IgG Elispots were used to measure the survival and functionality of the ASC. Contents of the EV fractions were evaluated by proteomics. We saw that both irradiated and non-irradiated MSC secretome preparations afforded vesicles of a size consistent with EVs. Both preparations appeared comparable in EM morphology and CD63 and CD81 immuno-gold EM. Both irradiated and non-irradiated EV fractions supported ASC function, at 88% and 90%, respectively, by day 3. In contrast, conventional media maintained only 4% ASC survival by day 3. To identify the specific factors that provided ASC support, we compared proteomes of the irradiated and non-irradiated EV fractions with conventional media. Pathway analysis of these proteins identified factors involved in the vesicle-mediated delivery of integrin signalling proteins. These findings indicate that BM-MSC EVs provide an effective support system for ASC survival and IgG secretion.
来自骨髓(BM)间充质基质细胞(BM-MSC)的细胞外囊泡(EVs)是细胞间短距离和长距离通讯的新机制。已证明BM-MSC可支持人抗体分泌细胞(ASC)的存活,但MSC与ASC相互作用之间的串扰是否可通过EVs发生尚不清楚。因此,我们评估了EVs在ASC存活和IgG分泌中的作用。从受辐照和未受辐照的原代BM-MSC中分离出EVs并进行定量。通过电子显微镜(EM)以及CD63和CD81免疫金EM染色对其进行进一步表征。通过荧光激活细胞分选(FACS)分离出人ASC,并与EV级分、EV减少级分或常规培养基一起培养。使用IgG酶联免疫斑点法测量ASC的存活和功能。通过蛋白质组学评估EV级分的含量。我们发现,受辐照和未受辐照的MSC分泌组制剂均能产生大小与EVs一致的囊泡。两种制剂在EM形态以及CD63和CD81免疫金EM方面看起来具有可比性。到第3天,受辐照和未受辐照的EV级分分别以88%和90%的比例支持ASC功能。相比之下,到第3天常规培养基仅维持4%的ASC存活。为了确定提供ASC支持的特定因素,我们将受辐照和未受辐照的EV级分的蛋白质组与常规培养基进行了比较。对这些蛋白质的通路分析确定了参与整合素信号蛋白囊泡介导递送的因素。这些发现表明,BM-MSC EVs为ASC存活和IgG分泌提供了有效的支持系统。