InVivo Bioluminescence Imaging of Transplanted Mesenchymal Stromal Cells and Their Rejection Mediated by Intrahepatic NK Cells
InVivo Bioluminescence Imaging of Transplanted Mesenchymal Stromal Cells and Their Rejection Mediated by Intrahepatic NK Cells
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DOI：
10.1007/s11307-016-0962-9
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发表时间：
2017-02
影响因子：
3.1
通讯作者：
Jing-jing Liu;Xiao-jun Hu;Zheng-ran Li;Rong-hua Yan;Dan Li;Jin Wang;H. Shan
中科院分区：
文献类型：
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作者：
Jing-jing Liu;Xiao-jun Hu;Zheng-ran Li;Rong-hua Yan;Dan Li;Jin Wang;H. Shan
目的间充质干细胞（MSCs）在肝脏疾病的治疗中具有广阔的应用前景。然而，肝内移植后MSCs存活时间短限制了其价值，因此，了解MSCs存活和排斥反应的基础可能会增加其效用。本研究采用Luc 2-mKate 2双融合报告基因（MSCs-R）标记人MSCs，通过活体生物发光成像（BLI）检测MSCs在肝内移植后的滞留时间和存活情况，探讨肝内NK细胞对MSCs存活和在肝内滞留的影响。进行MSC和NK细胞的共培养以评估细胞毒性。为了评价NK细胞在异种移植排斥反应中的作用，在肝内NK细胞激活后，用BLI检测移植的MSCs-R在体内的命运。（R2= 0.9956）。在体内，我们观察到在对照组和NK细胞组中，移植的MSCs-R在对应于肝脏的区域上的生物发光信号逐渐下降。激活组。然而，与对照组相比，NK活化组小鼠的存活时间和肝内MSCs-R的保留下降得更快。这表明激活的NK细胞加速了移植的MSC的消除。此外，我们发现肝内移植MSCs后，肝脏NK细胞的数量和NK活化标志物的表达显著增加。这表明，驻留的NK细胞，在静息状态下，被激活的肝内移植的人MSC。总之，数据表明，激活的肝脏NK细胞介导部分MSC异种移植物的排斥反应。细胞毒性实验表明，活化的NK细胞可抑制MSCs的增殖，并在一定程度上诱导MSCs死亡。结论BLI可在体内动态追踪人MSCs，小鼠肝脏NK细胞尤其是活化的NK细胞的作用可从MSCs信号丢失推断。这一发现可能对骨髓间充质干细胞移植治疗肝脏疾病具有实际的临床意义。
PurposeMesenchymal stromal cells (MSCs) hold promise in the treatment of liver disease. However, short survival time of MSCs after intrahepatic transplantation limits their value; therefore, understanding the basis of MSCs survival and rejection may increase their utility. This study was aimed at determining the role of intrahepatic natural killer (NK) cells on MSCs survival and their retention in the liver shortly after transplant.ProceduresHuman MSCs were labeled with the Luc2-mKate2 dual-fusion reporter gene (MSCs-R), and the residence time and survival of MSCs-R xenografts after intrahepatic transplantation were evaluated byin vivobioluminescence imaging (BLI). Coculture of MSCs and NK cells was performed to assess cytotoxicity. To evaluate the role of NK cells in rejection of the xenografted cells, the fates of transplanted MSCs-R were then assessedin vivoby BLI after activation of intrahepatic NK cells.ResultsWe observed a linear correlation between luciferase activity from live MSCs-R and cell numberin vitro(R2= 0.9956).In vivo, we observed a gradual decline in bioluminescent signals from transplanted MSCs-R over a region corresponding to the liver in both the control group and the NK-activated group. However, the survival time and retention of intrahepatic MSCs-R decreased more rapidly in the NK-activated group of mice compared to the control group. This indicated that activated NK cells accelerate the elimination of transplanted MSCs. Also, we found that the number of hepatic NK cells and the expression of NK activation markers significantly increased after intrahepatic delivery of MSCs. This suggested that resident NK cells, in a resting state, were activated by intrahepatic transplantation of human MSCs. Taken together, the data suggests that activated hepatic NK cells mediate, in part, rejection of the MSCs xenografts. Cytotoxicity assays showed that activated NK cells may inhibit the proliferation of MSCs and, to a certain extent, induce MSCs death.ConclusionHuman MSCs could be followed dynamicallyin vivoby BLI, and the role of murine hepatic NK cells, especially activated NK cells, could be inferred from the loss of signals from MSCs. This finding may have practical clinical implications in MSCs transplantation in treating liver disease.