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A comparison of three methods of decellularization of pig corneas to reduce immunogenicity

International Journal of Ophthalmology(English edition) 2014年第4期7卷 587-593页

作者： Whayoung Lee Yuko Miyagawa cassandra Long david k.c.cooper Hidetaka Hara Department of Surgery Thomas E. Starzl Transplantation Institute University of Pittsburgh

·AIM: To investigate whether decellularization using different techniques can reduce immunogenicity of the cornea, and to explore the decellularized cornea as a scaffold for cultured corneal endothelial cells(cEcs).Transplantation of decellularized porcine corneas increases graft transparency and survival for longer periods compared with fresh grafts.·METHODS: Six-month-old wild-type pig corneas were cut into 100-200 μm thickness, and then decellularized by three different methods: 1) 0.1% sodium dodecyl sulfate(SDS); 2) hypoxic nitrogen(N2); and 3) hypertonic Nacl. Thickness and transparency were assessed visually. Fresh and decellularized corneas were stained with hematoxylin/eosin(H&E), and for the presence of galactose-α1,3-galactose(Gal) and N-glycolylneuraminic acid(NeuGc, a nonGal antigen). Also, a human IgM/IgG binding assay was performed. cultured porcine cEcs were seeded on the surface of the decellularized cornea and examined after H&E staining.· RESULTS: All three methods of decellularization reduced the number of keratocytes in the stromal tissue by >80% while the collagen structure remained preserved. No remaining nuclei stained positive for Gal or NeuGc, and expression of these oligosaccharides on collagen was also greatly decreased compared to expression on fresh corneas. Human IgM/IgG binding to decellularized corneal tissue was considerably reduced compared to fresh corneal tissue. The cultured cEcs formed a confluent monolayer on the surface of decellularized tissue.· cONcLUSION: Though incomplete, the significant reduction in the cellular component of the decellularized cornea should be associated with a significantly reduced in vivo immune response compared to fresh corneas.

关键词： cornea decellularization immuneresponse pig xenotransplantation

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Genetically-engineered pig-to-human organ transplantation:a new beginning

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Science Bulletin 2022年第18期67卷 1827-1829页

作者： Xuan Zhang david k.c.cooper kefeng Dou Department of Hepatobiliary Surgery Xijing HospitalFourth Military Medical UniversityXi’an 710032China center for Transplantation Sciences Department of SurgeryMassachusetts General Hospital/Harvard Medical SchoolBoston 02114USA

There has been a continuing inadequate number of human organs for the treatment of patients with terminal organ failure *** transplantation of organs from genetically engineered pigs may prove an alternative solution[1].In recent years,great progress has been made in pig-to-nonhuman primate(NHP)organ transplantation models(Table S1 online).

关键词：器官移植 organs 1]

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The potential of genetically-engineered pigs in providing an alternative source of organs and cells for transplantation

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The Journal of Biomedical Research 2013年第4期27卷 249-253页

作者： david k.c.cooper Hidetaka Hara Mohamed Ezzelarab Rita Bottino Massimo Trucco carol Phelps david Ayares Yifan Dai Thomas E.Starzl Transplantation Institute University of Pittsburgh Medical Center Division of Immunogenetics Children's Hospital of Pittsburgh University of Pittsburgh Revivicor Inc Jiangsu key Laboratory of Xenotransplantation Nanjing Medical University

There is a critical shortage of organs, cells, and corneas from deceased human donors worldwide. There are also shortages of human blood for transfusion. A potential solution to all of these problems is the transplantation of organs, cells, and corneas from a readily available animal species, such as the pig, and the transfusion of red blood cells from pigs into humans. However, to achieve these ends, major immunologic and other barriers have to be overcome. considerable progress has been made in this respect by the genetic modification of pigs to protect their tissues from the primate immune response and to correct several molecular incompatibilities that exist between pig and primate. These have included knockout of genes responsible for the expression of major antigenic targets for primate natural anti-pig antibodies, insertion of human complement- and coagulation-regulatory transgenes, and knockdown of swine leukocyte antigens that stimulate the primate＇s adaptive immune response. As a result of these manipulations, the administration of novel immunosuppressive agents, and other innovations, pig hearts have now functioned in baboons for 6-8 months, pig islets have maintained normoglycemia in diabetic monkeys for 〉 1 year, and pig corneas have maintained transparency for several months. clinical trials of pig islet trans- plantation are already in progress. Future developments will involve further genetic manipulations of the organ- source pig, with most of the genes that are likely to be beneficial already identified.

关键词： pig blood transfusion pig genetic-engineered pig islets pig organs xenotransplantation

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PLATELET AGGREGATION IN HUMANS AND NONHUMAN PRIMATES:RELEVANcE TO XENOTRANSPLANTATION

PLATELET AGGREGATION IN HUMANS AND NONHUMAN PRIMATES:RELEVAN...

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2012中国器官移植大会

作者： Hayato Iwase Burcin Ekser Eefje Dons david k.c.cooper Mohamed Ezzelarab Thomas E Starzl Transplantation Institute University of PittsburghPittsburghPAUSA Department of Surgery Transplantation and Advanced TechologiesVascular Surgery and Organ Transplant UnitUniversity Hospital of CataniaCataniaItaly

【Introduction】Platelet activation/aggregation plays a key role in the dysregulation of coagulation and the development of thrombotic microangiopathy in nonhuman primate recipients of pig *** a preliminary to the study of anti-platelet therapy in vitro and in vivo,the present study aimed to compare platelet aggregation in whole blood from humans,baboons,and cynomolgus monkeys.【Methods】Using ’chrono-log’ technology(two-sample four-channel chrono-log Whole Blood Aggregometer),we studied

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hcD47诱导人巨噬细胞对猪内皮细胞的免疫耐受

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器官移植 2019年第2期10卷 165-170页

作者：胡文宝潘登科 davidk.c.cooper 蔡志明牟丽莎中山大学中山医学院广州510080 深圳市第二人民医院深圳大学第一附属医院中国农业科学院北京畜牧兽医研究所美国阿拉巴马大学伯明翰分校

目的探讨人cD47(hcD47)在诱导人巨噬细胞对猪内皮细胞免疫耐受中的作用。方法将转染了pcDH-hcD47-FLAG质粒的猪髂总动脉内皮细胞(PIEc)设为pcDH-hcD47组;将转染了pcDH-FLAG空载体质粒的PIEc为pcDH组;将转染了hcD47-dN的PIEc设为pcDH-hcD... 详细信息

目的探讨人cD47(hcD47)在诱导人巨噬细胞对猪内皮细胞免疫耐受中的作用。方法将转染了pcDH-hcD47-FLAG质粒的猪髂总动脉内皮细胞(PIEc)设为pcDH-hcD47组;将转染了pcDH-FLAG空载体质粒的PIEc为pcDH组;将转染了hcD47-dN的PIEc设为pcDH-hcD47-dN组;将人脐静脉内皮细胞(HUVEc)设为阳性对照组。将细胞分别与人巨噬细胞共培养,检测信号调节蛋白α(SIRPα)的磷酸化情况以及人巨噬细胞对PIEc的杀伤作用。进一步从GT-/-及GT-/-/hcD47基因编辑猪中分离了猪主动脉血管内皮细胞(PAEc),并分析SIRPα的磷酸化情况及人巨噬细胞对PAEc的杀伤作用。结果 pcDH组细胞不能诱导SIRPα的磷酸化,而pcDH-hcD47组细胞与人巨噬细胞共培养10 min后便能够激活SIRPα的磷酸化,且随着共培养时间的延长,SIRPα的磷酸化程度也随之增强。pcDH-hcD47-dN组细胞不能激活SIRPα的磷酸化。人巨噬细胞对p cDH组细胞产生了明显的杀伤作用,p cDH-h cD47组细胞可明显抑制人巨噬细胞的杀伤作用(PcDH-hcD47-dN组细胞则不能抑制人巨噬细胞的杀伤作用。GT-/--PAEc与人巨噬细胞共培养后,不能激活SIRPα的磷酸化;而GT-/-/hcD47-PAEc与人巨噬细胞共培养后则明显激活了SIRPα的磷酸化。人巨噬细胞对GT-/--PAEc产生了明显的杀伤作用,GT-/-/hcD47-PAEc可明显抑制人巨噬细胞的杀伤作用(PcD47可以通过激活SIRPα的磷酸化抑制人巨噬细胞对内皮细胞的杀伤作用。

关键词：异种移植 cD47 信号调节蛋白α 人巨噬细胞内皮细胞磷酸化免疫排斥反应

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异种移植免疫障碍综述

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现代医药卫生 2017年第7期33卷 1032-1035页

作者：高冰思周明澍 david k.c.cooper 黄余良王毅南华大学附属第二医院泌尿外科湖南衡阳421001 南华大学附属第二医院妇产科湖南衡阳421001 美国宾夕法尼亚州匹兹堡大学托马斯移植研究所 15261

近数十余年来,由于器官供体和受体之间的巨大缺口,cooper等团队致力于以基因工程猪为供体的异种器官移植研究。然而,人和非灵长类动物体内天然的抗猪抗体与表达与供体猪细胞表面的碳水化合物抗原（以α-1,3半乳糖为主）结合,激活补体级... 详细信息

近数十余年来,由于器官供体和受体之间的巨大缺口,cooper等团队致力于以基因工程猪为供体的异种器官移植研究。然而,人和非灵长类动物体内天然的抗猪抗体与表达与供体猪细胞表面的碳水化合物抗原（以α-1,3半乳糖为主）结合,激活补体级联反应,导致被移植的器官在几分钟内或者几小时内损伤,这一过程被称之为超急性免疫排斥反应。

关键词：器官移植移植,异种移植物排斥补体系统蛋白质类人凝血调节蛋白综述

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A SYNDROME OF SEVERE HYPOGLYcEMIA AND AcIDOSIS IN YOUNG IMMUNOSUPPRESSED DIABETIc MONkEYS - ASSOcIATION WITH SEPSIS

A SYNDROME OF SEVERE HYPOGLYCEMIA AND ACIDOSIS IN YOUNG IMMU...

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2012中国器官移植大会

作者： Dirk J.van der Windt Eefje M.Dons Gabriel J. Echeverri Noriko Murase Rita Bottino Martin Wijkstrom Robert Wagner david k.c.cooper Thomas E.Starzl Transplantation Institute Department of SurgeryUniversity of Pittsburgh Medical CenterPittsburghPAUSA Division of Immunogenetics Department of PediatricsChildren's Hospital of PittsburghPittsburghPAUSA

Large animals treated with immunosuppressive drug regimens for preclinical experiments of transplantation have increased risks of infection,as do individuals with *** we report our experience with severe sepsis in two young cynomolgus monkeys that were subjected to diabetes induction +/- islet *** both monkeys,infection was associated with a syndrome of profound hypoglycemia accompanied by severe *** the discussion of these cases we include information on five immunosuppressed pigs that also experienced severe hypoglycemic episodes after diabetes induction and islet

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Exercise immunology:Novel insights

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Journal of Sport and Health Science 2024年第3期13卷 277-279,I0001页

作者： david c.Nieman Human Performance Laboratory Appalachian State UniversityNorth Carolina Research CampusKannapolisNC 28081USA

The coronavirus disease 2019(cOVID-19)pandemic heightened worldwide attention on the influence of exercise on the immune *** were raised about whether regular moderate-to-vigorous physical activity(MVPA)could help prevent cOVID-19 or mitigate symptoms when *** MVPA bolster immunity among those most prone to cOVID-19 including the elderly,obese,and immunocompromised patients?Does regular MVPA dampen systemic inflammation,thereby lessening the cOVID-19 cytokine storm?

关键词： immunity inflammation thereby

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Artificial intelligence analysis of videos to augment clinical assessment:an overview

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Neural Regeneration Research 2024年第4期19卷 717-718页

作者： david c.Wong Stefan Williams University of Leeds Leeds Teaching Hospitals Trust LeedsUK

Observation is a fundamental part of the practice of clinical *** of movement is particularly important for the *** such as Parkinson’s disease,multiple sclerosis,stroke,epilepsy,and many others affect a person’s movement in characteristic *** some conditions,changes in the patient’s voice can be included in this-changes in sound caused by changes in the movements of *** clinician’s detection of a characteristic abnormality,and their judgment of its severity,plays a central role in both diagnosis and the assessment of prognosis or response to ***,that practice depends upon a limited resource of experienced *** addition,these experts are limited by human visual judgment,which cannot reliably or precisely detect and measure small or subtle changes in movement(Williams et al.,2023).

关键词： clinical epilepsy diagnosis

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Balancing machine learning and artificial intelligence in soil science with human perspective and experience

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Pedosphere 2024年第1期34卷 9-12页

作者： david c.WEINDORF Somsubhra cHAkRABORTY Department of Earth and Atmospheric Science Central Michigan Universityount Pleasant M148859USA Agricultural and Food Engineering Department Indian Institute of TechnologyKharagpur 721302India

Machine learning and artificial intelligence continue to evolve at a rapid pace, with many potential applications related to soil science. Even so, human experience and perception play an invaluable role in characterizing soil properties, especially qualitative properties that may elude sensing/computer-based modeling approaches. The elegant solution to this conundrum relies on the synthesis of computer-aided predictive modeling with human insight and *** global population surpasses 8 billion, the importance of optimized agronomic production to feed a hungry world has never been more important.

关键词： soil elude qualitative

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