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In vivo anti-tumor activity of murine hematopoietic stem cells expressing a p185HER2-specific chimeric t-cell receptor gene

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Journal of Microbiology and Immunology 2006年第2期4卷 117-124页

作者： JIAN MIN YANG MICHAEL S FRIEDMAN MARIANNE t HUBEN JENNIFER FULLER QIAO LI ALFRED E CHANG JAMES J MULE kevin t mcdonagh Department of Hematology Changhai Hospital the Second Military Medical University Shanghai P. R. China Department of Internal Medicine University of Michigan Medical School Ann Arbor Michigan USA Department of Surgery University of Michigan Medical School Ann Arbor Michigan USA Division of Hematology Oncology and Blood and Marrow Transplantation Department of Internal Medicine University of Kentucky College of Medicine Lexington Kentucky USA

We have confirmed efficient anti-tumor activities of the peripheral lymphocytes transduced with a p185HEH2-specific chimeric t-cell receptor gene both in murine and in human in our previous studies. to further test the feasibility of chimeric t-cell receptor in a bone marrow transplantation model, we first, made two routine tumor cell lines： Mt901 and MCA-205, to express human p185HER2 by retroviral gene transduction. Murine bone marrow cells were retrovirally transduced to express the chimeric t-cell receptor and gene-modified bone marrow cells were transplanted into lethally irradiated mouse. Six months post transplantation, p185HER2-positive tumor ceils： Mt-901/HER2 or MCA-205/ HER2 was subcutaneously or intravenously injected to make mouse models simulating primary breast cancer or pulmonary metastasis. the in vivo anti-tumor effects were monitored by the size of the subcutaneous tumor or counting the tumor nodules in the lungs after India ink staining. the size of the subcutaneous tumor was significantly inhibited and the number of pulmonary nodules were significantly decreased in mouse recipients transplanted with chimeric t-cell receptor modified bone marrow cells compared with the control group. Our results suggest the efficient in vivo anti-tumor activities of chimeric t-cell receptor gene modified bone marrow cells.

关键词： Gene therapy Retrovirus Chimeric t-cell receptor Murine Hematopoietic stem cell Immunotherapy

嵌合t细胞受体N29γ的t细胞靶向性治疗p185HER2阳性鼠乳腺癌肺转移

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癌症 2004年第z1期23卷 1370-1375页

作者：李升平 Frank A.Urban Jennifer N.MacGregor Dennis P.Hughes kevin t.mcdonagh 中山大学肿瘤防治中心肝胆科广东广州5100602 Department of Internal Medicine Hematology/OncologyUniversity of Michigan Medical SchoolAnn ArborMIUSA

背景和目的:嵌合t细胞受体(chimerict-cellreceptor,chtCR)是通过基因工程技术重组的免疫受体。它既具有与单克隆抗体相似的特异性结合抗原的特性,又能激活t细胞。与“经典”的tCR相比,chtCR介导的t细胞对靶细胞的作用的一个重要的优势... 详细信息

背景和目的:嵌合t细胞受体(chimerict-cellreceptor,chtCR)是通过基因工程技术重组的免疫受体。它既具有与单克隆抗体相似的特异性结合抗原的特性,又能激活t细胞。与“经典”的tCR相比,chtCR介导的t细胞对靶细胞的作用的一个重要的优势是不受MHC的限制和不需抗原的提呈过程。N29γ是特异性识别和结合p185HER2的嵌合t细胞受体。本研究将探讨表达N29γ的t细胞对HER2阳性的小鼠乳腺癌肺转移的靶向性治疗作用。方法:尼龙柱纯化的Balb/c小鼠脾脏t淋巴细胞,经小鼠CD3和CD28单克隆抗体激活,用离心法将pRet6N29γ转导入t细胞,流式细胞仪检测对照组绿色荧光蛋白(GFP)的表达以间接估计chtCRN29γ转导率。将荷有Mt901或Mt901/HER2瘤3天或8天的Balb/c小鼠随机分为治疗组和对照组,从尾静脉分别注入转导了N29γ或GFP逆转录病毒载体的t细胞(5×106～40×106/只),每只小鼠腹腔内注入重组人白细胞介素Ⅱ3×104IU,每12h一次,共10次。t细胞治疗后11天(荷瘤3天动物模型)或13天(荷瘤8天动物模型)处死动物,计数肺转移肿瘤数目。结果:在荷瘤3天的动物模型中,肺转移瘤数目在空白对照组、无病毒载体转导的t细胞治疗组、转导有GFP的t细胞治疗组均大于200个/肺;表达N29γchtCR的t细胞能使Mt901/HER2肺转移瘤消退(3.4±3.3)个/肺,但?

关键词：嵌合t细胞受体乳腺癌 HER2/neu 转移

tumor Antigen Specific Activation of Primary Human t-Cells Expressing a Virally Encoded Chimeric t-Cell Receptor Specific for p185HER2

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Journal of Microbiology and Immunology 2004年第4期2卷 272-277页

作者：杨建民 Michael S FRIEDMAN Christopher M REYNOLDS Marianne t HUBEN Lee WILKE Jennifer FULLER 李桥 Zelig ESHHAR James J MULE kevin t mcdonagh Department of Hematology Changhai Hospital The Second Millitay Medical University Shanghai 200433 China Departments of Internal Medicine Departments of Surgery Departments of Internal Medicine University of Michigan Medical School Ann Arbor MI 48109 and De- partment of Chemical Immunology the Weizmann Institute of Science Israel

We have developed and tested chimeric t-cell receptors (tCR) specific for p185HER2. In these experiments, retroviral vectors expressing the N29γ or N29ζ receptors were constructed in pREt6. Amphotropic viral producer cells were established in the GALV-based PG13 packaging cell line. Ficoll purified human peripheral blood lymphocytes (PBL) were virally transduced using an optimized protocol incorporating activation with immobilized anti-CD3/anti-CD28 monoclonal anti- bodies, followed by viral infection in the presence of fibronectin fragment CH296. transduced cells were co-cultured with human tumor cell lines that overexpress (SK-OV-3) or underexpress (MCF7) p185HER2 to assay for antigen specific im- mune responses. Both CM+ and CD8+ t-cells transduced with the N29γ or N29ζ chtCR demonstrated HER2-specific anti- gen responses, as determined by release of th1 like cytokines, and cellular cytotoxicity assays. Our results support the fea- sibility of adoptive immunotherapy with genetically modified t-cells expressing a chtCR specific for p185HER2.

关键词： Gene therapy retrovirus Chimeric t-cell receptor Human peripheral t-lymphocytes Immunotherapy

转染嵌合性t细胞受体基因小鼠t淋巴细胞的体外抗肿瘤作用

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中国肿瘤生物治疗杂志 2006年第4期13卷 243-248页

作者：杨建民 Michael S Friedman 李峤 James J Mule Alfred E Chang kevin t mcdonagh 第二军医大学长海医院血液内科 University of Michigan Medical SchoolAnn Arbor MI 48109-0620 USA University of Kentucky College of Medicine Lexington Ky 40536-0293 USA

目的:研究小鼠t淋巴细胞在转染嵌合性t细胞受体基因后的体外抗肿瘤作用。方法:应用重组DNA技术,将HER2特异性嵌合性t-细胞受体构建入逆转录病毒载体;转入包装细胞后,收集病毒上清,转染小鼠t淋巴细胞,转基因后的小鼠t淋巴细胞分别与HER2... 详细信息

目的:研究小鼠t淋巴细胞在转染嵌合性t细胞受体基因后的体外抗肿瘤作用。方法:应用重组DNA技术,将HER2特异性嵌合性t-细胞受体构建入逆转录病毒载体;转入包装细胞后,收集病毒上清,转染小鼠t淋巴细胞,转基因后的小鼠t淋巴细胞分别与HER2阳性(SK-OV-3)或阴性(MCF-7)的肿瘤细胞系共培养,检测其细胞因子γ干扰素释放,51Cr释放法检测CtL评价其抗肿瘤效应。结果:所构建载体经酶切鉴定符合要求,乒乓法转染包装细胞系GP+E86,检测病毒滴度为1.2×106,Retronectin结合离心法转染经抗CD3/CD28单抗活化的小鼠t淋巴细胞,转染效率可达50%以上;转染嵌合性t细胞受体基因的t淋巴细胞与HER2阳性或阴性的肿瘤细胞系共培养后可检测到HER2特异性的细胞因子γ干扰素释放,51Cr释放法测CtL可见转染嵌合性t细胞受体基因t淋巴细胞对HER2阳性的肿瘤细胞具显著杀伤效应。结论:转染嵌合性t细胞受体基因的小鼠t淋巴细胞在体外可通过细胞因子释放和CtL效应发挥显著的抗肿瘤作用。

关键词：嵌合性t细胞受体基因转染免疫治疗小鼠 t淋巴细胞

An exciting time for genome editing: an interview with David R. Liu

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National Science Review 2019年第3期6卷 452-454页

作者： Weijie Zhao kevin t.Zhao NSR David Liu’s group at Harvard University and the Broad Institute

In 1987, several Osaka University researchers discovered a special kind of clustered DNA repeats in bacteria. Within a few years, two other groups independently discovered the same phenomenon but no one knew its function at the time. Only a small handful of scientists studied this property from its discovery in 1987 to 2005. It was then that the function of these DNA repeats, which were named Clustered

关键词： pro CRISPR Liu an interview with David R An exciting time for genome editing

Surgical site soft tissue thickness as a predictor of complications following arthroplasty

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World Journal of Methodology 2025年第2期15卷 62-73页

作者： kevin A Wu Faheem Pottayil Crystal Jing Ankit Choudhury Albert t Anastasio Department of Orthopaedic Surgery Duke University HospitalDurhamNC 27710United States Department of Orthopaedic Surgery Medical College of Georgia at Augusta UniversityAugustaGA 30912United States Department of Orthopaedic Surgery Medical College of WisconsinMilwaukeeWI 53226United States

Appreciation of soft-tissue thickness(Stt)at surgical sites is an increasingly recognized aspect of arthroplasty procedures as it may potentially impacting postoperative *** research has focused on the predictive value of preoperative Stt measurements for complications following various forms of arthroplasty,particularly infections,across procedures such as total knee,hip,shoulder,and ankle *** studies have indicated that increased Stt is associated with a higher risk of complications,including infection and wound healing *** assessment of Stt before surgery could play a crucial role in identifying patients at a higher risk of complications and may be instru-mental in guiding preoperative planning to optimize outcomes in arthroplasty *** measurement techniques and further research are essential to enhance the reliability and clinical utility of Stt assessment for arthro-plasty surgery.

关键词： Soft-tissue thickness Arthroplasty Surgical complications total knee arthroplasty total hip arthroplasty total shoulder arthroplasty total ankle arthroplasty Preoperative assessment Wound healing Infection risk

A FORGOttEN BOOK?HOW A 36-YEAR-OLD BOOK INFLUENCED GREEN DESIGN

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Journal of Green Building 2013年第2期8卷 44-53页

作者： kevin t.Perry Murray State University MurrayKentucky

INtRODUCtION the list of books and references on green building and sustainable development has grown tremendously over the last decade and a *** of this has coincided with the growth of the LEED and other green rating systems,as well as society’s realization about the benefits and potential of sustainable *** back though,there were many wonderful and influential texts written around the time of the 1970s’oil/energy crisis that may have never been seen by today’s *** article examines one of those important texts,A Pattern Language:towns,Buildings,Construction,and briefly outlines how it influenced some of the green practices seen today.

关键词： A Pattern Language books CNU green building LEED neighborhood development sustainable development

Aqueous ruthenium detection by microwave-assisted laser-induced breakdown spectroscopy

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Plasma Science and technology 2022年第8期24卷 32-39页

作者： Abu Farhan ABU KASIM M A WAKIL kevin GRANt Milton HEARN Zeyad t ALWAHABI School of Chemical Engineering The University of AdelaideAdelaide SA 5005Australia

Aqueous ruthenium was detected in real-time under ambient conditions using microwaveassisted laser-induced breakdown spectroscopy(MW-LIBS).A 10 mJ laser energy and 750 W microwave power were directed at an open liquid jet sample of *** was observed that,for liquid flow,the coupling efficiency between the microwave and the laser-induced plasma was limited to 43%.the improvement in the ruthenium’s signal-to-noise ratio with MW-LIBS,with respect to LIBS,was *** on MW-LIBS,the limit of detection for aqueous ruthenium was determined to be 957±84 ppb.

关键词： microwave-assisted laser induced breakdown spectroscopy ruthenium detection laser induced breakdown spectroscopy

An Interactive Visual Analytics Framework for Multi-Field Data in a Geo-Spatial Context

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tsinghua Science and technology 2013年第2期18卷 111-124页

作者： Zhiyuan Zhang Xiaonan tong kevin t. McDonnell Alla Zelenyuk Dan Imre Klaus Mueller the Visual Analytics and Imaging Lab at Computer Science Department Stony Brook University Stony Brook NY 11790 USA an undergraduate student at Stanford University Stanford CA 94305 USA the Department of Mathematics Dowling College Oakdale NY 11769 USA Pacific Northwest National Lab Richland WA 99354 USA also with SUNY Korea

Climate research produces a wealth of multivariate data. these data often have a geospatial reference and so it is of interest to show them within their geospatial context. One can consider this configuration as a multifield visualization problem, where the geo-space provides the expanse of the field. However, there is a limit on the amount of multivariate information that can be fit within a certain spatial location, and the use of linked multivariate information displays has previously been devised to bridge this gap. In this paper we focus on the interactions in the geographical display, present an implementation that uses Google Earth, and demonstrate it within a tightly linked parallel coordinates display. Several other visual representations, such as pie and bar charts are integrated into the Google Earth display and can be interactively manipulated. Further, we also demonstrate new brushing and visualization techniques for parallel coordinates, such as fixed-window brushing and correlation-enhanced display. We conceived our system with a team of climate researchers, who already made a few important discoveries using it. this demonstrates our system＇s great potential to enable scientific discoveries, possibly also in other domains where data have a geospatial reference.

关键词： geospatial visualization visual analytics information visualization multivariate visualization parallel coordinates coordinated displays linking and brushing