Conradi Lenard, Schmidt Stephanie, Neofytou Evgenios, Deuse Tobias, Peters Laura, Eder Alexandra, Hua Xiaoqin, Hansen Arne, Robbins Robert C, Beygui Ramin E, Reichenspurner Hermann, Eschenhagen Thomas, Schrepfer Sonja
University Heart Center Hamburg, Transplant and Stem Cell Immunobiology Laboratory, Hamburg, Germany; Department of Cardiovascular Surgery, University Heart Center Hamburg, Hamburg, Germany; DZHK (German Center for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, and Cardiovascular Research Center and Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; Stanford Cardiovascular Institute and Department of Cardiothoracic Surgery, Stanford University School of Medicine, Falk Cardiovascular Research Center, Stanford, California, USA.
University Heart Center Hamburg, Transplant and Stem Cell Immunobiology Laboratory, Hamburg, Germany; Department of Cardiovascular Surgery, University Heart Center Hamburg, Hamburg, Germany; DZHK (German Center for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, and Cardiovascular Research Center and Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; Stanford Cardiovascular Institute and Department of Cardiothoracic Surgery, Stanford University School of Medicine, Falk Cardiovascular Research Center, Stanford, California, USA
Stem Cells Transl Med. 2015 Jun;4(6):625-31. doi: 10.5966/sctm.2013-0202. Epub 2015 May 6.
Different tissue-engineering approaches have been developed to induce and promote cardiac regeneration; however, the impact of the immune system and its responses to the various scaffold components of the engineered grafts remains unclear. Fibrin-based engineered heart tissue (EHT) was generated from neonatal Lewis (Lew) rat heart cells and transplanted onto the left ventricular surface of three different rat strains: syngeneic Lew, allogeneic Brown Norway, and immunodeficient Rowett Nude rats. Interferon spot frequency assay results showed similar degrees of systemic immune activation in the syngeneic and allogeneic groups, whereas no systemic immune response was detectable in the immunodeficient group (p < .001 vs. syngeneic and allogeneic). Histological analysis revealed much higher local infiltration of CD3- and CD68-positive cells in syngeneic and allogeneic rats than in immunodeficient animals. Enzyme-linked immunospot and immunofluorescence experiments revealed matrix-directed TH1-based rejection in syngeneic recipients without collateral impairment of heart cell survival. Bioluminescence imaging was used for in vivo longitudinal monitoring of transplanted luciferase-positive EHT constructs. Survival was documented in syngeneic and immunodeficient recipients for a period of up to 110 days after transplant, whereas in the allogeneic setting, graft survival was limited to only 14 ± 1 days. EHT strategies using autologous cells are promising approaches for cardiac repair applications. Although fibrin-based scaffold components elicited an immune response in our studies, syngeneic cells carried in the EHT were relatively unaffected.
An initial insight into immunological consequences after transplantation of engineered heart tissue was gained through this study. Most important, this study was able to demonstrate cell survival despite rejection of matrix components. Generation of syngeneic human engineered heart tissue, possibly using human induced pluripotent stem cell technology with subsequent directed rejection of matrix components, may be a potential future approach to replace diseased myocardium.
已经开发出不同的组织工程方法来诱导和促进心脏再生；然而，免疫系统及其对工程移植物各种支架成分的反应的影响仍不清楚。基于纤维蛋白的工程心脏组织（EHT）由新生Lewis（Lew）大鼠心脏细胞生成，并移植到三种不同大鼠品系的左心室表面：同基因Lew大鼠、异基因Brown Norway大鼠和免疫缺陷的Rowett Nude大鼠。干扰素斑点频率测定结果显示，同基因组和异基因组的全身免疫激活程度相似，而免疫缺陷组未检测到全身免疫反应（与同基因组和异基因组相比，p <.001）。组织学分析显示，同基因和异基因大鼠中CD3和CD68阳性细胞的局部浸润远高于免疫缺陷动物。酶联免疫斑点和免疫荧光实验显示，同基因受体中存在基于基质的TH1介导的排斥反应，而心脏细胞存活未受附带损害。生物发光成像用于对移植的荧光素酶阳性EHT构建体进行体内纵向监测。同基因和免疫缺陷受体在移植后长达110天的时间内记录到移植物存活，而异基因移植情况下，移植物存活仅限制在14±1天。使用自体细胞的EHT策略是心脏修复应用中有前景的方法。尽管在我们的研究中基于纤维蛋白的支架成分引发了免疫反应，但EHT中携带的同基因细胞相对未受影响。
通过本研究初步了解了工程心脏组织移植后的免疫后果。最重要的是，本研究能够证明尽管基质成分被排斥，但细胞仍能存活。利用人类诱导多能干细胞技术生成同基因人类工程心脏组织，随后定向排斥基质成分，可能是未来替代病变心肌的一种潜在方法。
