Immunological Properties of Murine Parthenogenetic Stem Cell-Derived Cardiomyocytes and Engineered Heart Muscle.
Immunological Properties of Murine Parthenogenetic Stem Cell-Derived Cardiomyocytes and Engineered Heart Muscle.
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DOI：
10.3389/fimmu.2017.00955
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发表时间：
2017
影响因子：
7.3
通讯作者：
Zimmermann WH
中科院分区：
文献类型：
--
作者：
Didié M;Galla S;Muppala V;Dressel R;Zimmermann WH
多能单性生殖干细胞（pSC）可以通过未受精卵母细胞的药理学激活来衍生。 pSC 中主要组织相容性复合体 (MHC) 的纯合性使其成为同种异体组织修复应用中有吸引力的细胞来源。最近在基于 pSC 的组织工程心脏修复中证实了这一点。然而，缺乏对 pSC 衍生的心肌细胞及其工程心肌（EHM）的免疫学特性的详细分析。本研究的目的是确定 pSC 衍生心肌细胞和 pSC-EHM 体内外的基线和细胞因子诱导的 I 类 MHC 和 II 类以及程序性死亡配体 1 (PDL-1) 和共刺激蛋白 (CD40、CD80、CD86) 的表达。通过利用最近开发的心肌细胞特异性遗传选择方案，来自 MHC 同源 (H2d/d) pSC 系的心肌细胞被富集至约 90%。心肌细胞中MHC I类和MHC II类表达只有在用干扰素γ（IFN-γ）刺激后才能观察到。 PDL-1 在 IFN-γ 作用下显着上调。 CD40、CD80 和 CD86 低水平表达，并且不被 IFN-γ 上调。由 H2d/d 心肌细胞构建的 EHM 在基础条件下表达同样低水平的 MHC I 类、MHC II 类和共刺激分子。然而，在 EHM 中，只有 MHC I 类分子在 IFN-γ 刺激后上调，而 MHC II 类分子则不然。接下来，我们采用 MHC 匹配和 MHC 不匹配的脾细胞和 T 细胞的共培养系统来分析 EHM 的免疫刺激特性。尽管存在 MHC 不匹配的条件，EHM 并未在体外诱导脾细胞或 T 细胞增殖。为了评估 pSC 衍生的心肌细胞在体内的免疫原性，我们在 MHC 匹配和不匹配小鼠的肾被膜下消除非心肌细胞（心体）后植入 pSC 衍生的胚状体。在匹配条件下可以观察到心体的自发跳动 28 天，在不匹配条件下可以观察到 7 天。畸胎瘤仅在 MHC 匹配的条件下 28 天后形成。免疫组织化学显示，在不匹配的条件下，植入物边缘区域存在单簇 CD3 阳性细胞，且很少有 CD3 阳性细胞浸润植入物。总而言之，MHC 匹配的 pSC 心肌细胞同种异体移植物显示出很少的免疫细胞激活，这为在没有免疫抑制的情况下观察到的 pSC-EHM 同种异体移植物的长期保留提供了解释。
Pluripotent parthenogenetic stem cells (pSCs) can be derived by pharmacological activation of unfertilized oocytes. Homozygosity of the major histocompatibility complex (MHC) in pSCs makes them an attractive cell source for applications in allogeneic tissue repair. This was recently demonstrated for pSC-based tissue-engineered heart repair. A detailed analysis of immunological properties of pSC-derived cardiomyocytes and engineered heart muscle (EHM) thereof is, however, lacking. The aim of this study was to determine baseline and cytokine-inducible MHC class I and MHC class II as well as programmed death ligand-1 (PDL-1) and co-stimulatory protein (CD40, CD80, CD86) expression in pSC-derived cardiomyocytes and pSC-EHM in vitro and in vivo. Cardiomyocytes from an MHC-homologous (H2d/d) pSC-line were enriched to ~90% by making use of a recently developed cardiomyocyte-specific genetic selection protocol. MHC class I and MHC class II expression in cardiomyocytes could only be observed after stimulation with interferon gamma (IFN-γ). PDL-1 was markedly upregulated under IFN-γ. CD40, CD80, and CD86 were expressed at low levels and not upregulated by IFN-γ. EHM constructed from H2d/d cardiomyocytes expressed similarly low levels of MHC class I, MHC class II, and costimulatory molecules under basal conditions. However, in EHM only MHC class I, but not MHC class II, molecules were upregulated after IFN-γ-stimulation. We next employed a cocultivation system with MHC-matched and MHC-mismatched splenocytes and T-cells to analyze the immune stimulatory properties of EHMs. Despite MHC-mismatched conditions, EHM did not induce splenocyte or T-cell proliferation in vitro. To evaluate the immunogenicity of pSC-derived cardiomyocytes in vivo, we implanted pSC-derived embryoid bodies after elimination of non-cardiomyocytes (cardiac bodies) under the kidney capsules of MHC-matched and -mismatched mice. Spontaneous beating of cardiac bodies could be observed for 28 days in the matched and for 7 days in the mismatched conditions. Teratomas formed after 28 days only in the MHC-matched conditions. Immunohistochemistry revealed single clusters of CD3-positive cells in the border zone of the implant in the mismatched conditions with few CD3-positive cells infiltrating the implant. Taken together, MHC-matched pSC-cardiomyocyte allografts show little immune cell activation, offering an explanation for the observed long-term retention of pSC-EHM allografts in the absence of immunosuppression.
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影响因子：
3.7
作者：
Baban B;Liu JY;Qin X;Weintraub NL;Mozaffari MS
通讯作者：
Mozaffari MS
小鼠多能孤雌胚胎干细胞衍生物构建组织工程皮肤等效物的临床前研究
影响因子：
7.5
作者：
Rao Y;Cui J;Yin L;Liu W;Liu W;Sun M;Yan X;Wang L;Chen F
通讯作者：
Chen F
影响因子：
15.9
作者：
Didie, Michael;Christalla, Peter;Zimmermann, Wolfram-Hubertus
通讯作者：
Zimmermann, Wolfram-Hubertus
影响因子：
15.9
作者：
Klug, MG;Soonpaa, MH;Field, LJ
通讯作者：
Field, LJ
影响因子：
14.8
作者：
Quah, Ben J. C.;Warren, Hilary S.;Parish, Christopher R.
通讯作者：
Parish, Christopher R.