■间充质基质细胞(MSC)的免疫调节特性已在体外和体内模型中得到充分表征。我们以前已经表明,肝MSC(L-MSC)是T细胞活化/增殖的优良抑制剂,NK细胞溶细胞功能,和巨噬细胞活化与体外脂肪(A-MSC)和骨髓MSC(BM-MSC)相比。
■为了在体内测试这些观察结果,我们将这些类型的MSC输注到单侧肾动脉狭窄(RAS)的小鼠中,建立肾脏炎症模型。11周龄通过剖腹手术诱发单侧RAS,雄性129-S1小鼠全身麻醉。对照小鼠进行假手术。人L-MSC,AMSC,手术后2周动脉内注射BM-MSC(各5x105个细胞)或PBS载体。输注后2周使用显微MRI成像研究肾脏形态。肾脏炎症,凋亡,纤维化,和MSC保留使用westernblot进行离体研究,免疫荧光,和免疫组织学分析。
■所有RAS小鼠的狭窄肾脏体积较小,确认重大伤害,并通过输注所有类型的MSC得到改善。与未处理的RAS相比,所有输注MSC的组具有较低水平的血浆肾素和蛋白尿。用BM-和L-MSC治疗的血清肌酐改善。位于狭窄肾脏并保留在狭窄肾脏内的所有类型的MSC,但L-MSC的保留率明显高于A-和BM-MSC。虽然所有组的MSC处理的小鼠表现出减少的整体炎症和巨噬细胞计数,L-MSC在定位于炎症部位和诱导M2(修复性)巨噬细胞极化以减少炎症变化方面显示出优异的体内效力。
■这些体内发现扩展了我们的体外研究,并表明L-MSC具有独特的抗炎特性,可能在肝脏诱导的耐受性中起作用,并进一步支持其用作具有潜在炎症病理生理学的疾病的治疗剂。
UNASSIGNED: The immunomodulatory properties of mesenchymal stromal cells (MSC) have been well-characterized in in-vitro and in-vivo models. We have previously shown that liver MSC (L-MSC) are superior inhibitors of T-cell activation/proliferation, NK cell cytolytic function, and macrophage activation compared to adipose (A-MSC) and bone marrow MSC (BM-MSC) in-vitro.
UNASSIGNED: To test these observations in-vivo, we infused these types of MSC into mice with unilateral renal artery stenosis (RAS), an established model of kidney inflammation. Unilateral RAS was induced via laparotomy in 11-week-old, male 129-S1 mice under general anesthesia. Control mice had sham operations. Human L-MSC, AMSC, and BM-MSC (5x105 cells each) or PBS vehicle were injected intra-arterially 2 weeks after surgery. Kidney morphology was studied 2 weeks after infusion using micro-MRI imaging. Renal inflammation, apoptosis, fibrosis, and MSC retention were studied ex-vivo utilizing western blot, immunofluorescence, and immunohistological analyses.
UNASSIGNED: The stenotic kidney volume was smaller in all RAS mice, confirming significant injury, and was improved by infusion of all MSC types. All MSC-infused groups had lower levels of plasma renin and proteinuria compared to untreated RAS. Serum creatinine improved in micetreated with BM- and L-MSC. All types of MSC located to and were retained within the stenotic kidneys, but L-MSC retention was significantly higher than A- and BM-MSC. While all groups of MSC-treated mice displayed reduced overall inflammation and macrophage counts, L-MSC showed superior potency in-vivo at localizing to the site of inflammation and inducing M2 (reparative) macrophage polarization to reduce inflammatory changes.
UNASSIGNED: These in-vivo findings extend our in-vitro studies and suggest that L-MSC possess unique anti-inflammatory properties that may play a role in liver-induced tolerance and lend further support to their use as therapeutic agents for diseases with underlying inflammatory pathophysiology.