Analysis of engraftment kinetics of donor-derived cells in multi-cord blood transplantation

多脐带血移植中供体来源细胞的植入动力学分析

• 批准号: 21591232
• 负责人: WASTANABE Nobukazu
• 金额: $ 2.91万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21591232/
• 关键词: 複数臍帯血移植; 生着不全; キメリズム解析; HLAミスマッチ; 抗HLA抗体; フローサイトメーター; キメリズム

Due to its many benefits, since 2003 the number of cord blood transplantations(CBT) being performed has increased to almost equal that of bone marrow transplantation in Japan. CBT, however, can be complicated by graft failure, which must be resolved for safe transplantation. To overcome these problems, double-unit cord blood transplantation(dCBT) has been developed. In Japan, a large scale clinical trial ran from 2006 to 2010 to check the effects of dCBT on the engraftment process. The results from this and other global dCBT clinical trials, however, are controversial. Additionally, basic research are not complete. Therefore, it is very important to investigate engraftment kinetics of dCBT and develop effective protocols on the basis of research data.At the start of this research project, we planned to analyze recipient blood after dCBT. After March 2010, however, there were no dCBT clinical trials in Japan. Therefore, we changed the project style from clinical investigation to analysi … More s of artificial chimerism samples in culture.In 2009, we had many biotinylated commercial allele specific HLA monoclonal antibodies and we discovered red blood cell contamination in the leukocyte fraction of peripheral blood. We found that previous multi-color chimerism analysis systems could not clearly analyze different kinds of HLA-bearing white blood cells from multiple persons.In 2010, we changed the combination of antibodies and fluorescent agents to obtain a good separation of each subpopulation and proper compensations. Finally, we successfully established an 11-color flow cytometry system for accurate analysis. We used PE-Cy5-conjugaed anti-CD235a antibody to get rid of red blood cells from gates on white blood cells for chimerism analysis. In addition, we made an anti-HLA-B62 antibody which stained cells very brightly.In 2011, we used our 11-color flow cytometry system to analyze triple chimerism in CD4^+T cells and CD8^+T cells in a mixed lymphocyte culture to measure frequencies of three donors' white blood cells and analyze mRNA levels of each person's CD4^4 T cells and CD8^+T cells which will be useful to understand engraftment kinetics and molecular basis of allo-reactivity in future dCBT. Less

由于其诸多好处，自2003年以来，在日本进行的脐带血移植（CBT）的数量增加到几乎与骨髓移植相同。然而，CBT可能因移植物衰竭而复杂化，这必须为安全移植而解决。为了克服这些问题，双单位脐带血移植（dCBT）得到了发展。在日本，从2006年到2010年进行了大规模的临床试验，以检查dCBT对植入过程的影响。然而，这项和其他全球dCBT临床试验的结果是有争议的。此外，基础研究也不完善。因此，研究dCBT的植入动力学并在研究数据的基础上制定有效的方案是非常重要的。在本研究项目开始时，我们计划对dct后的受者血液进行分析。然而，在2010年3月之后，日本没有进行dCBT临床试验。因此，我们将项目风格从临床调查转变为培养中人工嵌合样本的分析。在2009年，我们有许多生物素化的商业等位基因特异性HLA单克隆抗体，我们在外周血白细胞部分发现了红细胞污染。我们发现，以往的多色嵌合分析系统不能清楚地分析不同种类的人hla白细胞。2010年，我们改变了抗体和荧光剂的组合，以获得每个亚群的良好分离和适当的补偿。最后，我们成功地建立了一个11色流式细胞仪系统进行准确的分析。我们使用pe - cy5偶联抗cd235a抗体从白细胞上的门中去除红细胞进行嵌合分析。此外，我们制作了抗hla - b62抗体，使细胞染色非常明亮。2011年，我们使用11色流式细胞仪分析了混合淋巴细胞培养中CD4^+T细胞和CD8^+T细胞的三嵌合现象，测量了三个供者白细胞的频率，并分析了每个人CD4^4 T细胞和CD8^+T细胞的mRNA水平，这将有助于了解未来dCBT的植入动力学和同种异体反应性的分子基础。少

项目成果

Flow cytometric analysis of donor-recipient chimerism using anti-HLA antibody after HLA-mismatched hematopoietic cell and organ transplantation

HLA 不匹配的造血细胞和器官移植后使用抗 HLA 抗体对供体-受体嵌合现象进行流式细胞术分析

• 发表时间: 2010
• 作者: Stephanie C. Napier;Satoshi Yamazaki;Nobukazu Watanabe;Naofumi Matsuno;Satoshi Takahashi;Akinobu Takaki;and Hiromitsu Nakauchi
• 通讯作者: and Hiromitsu Nakauchi

11カラーFACS解析による造血細胞移植後の生着動態と白血病細胞の同時モニタリング

使用 11 色 FACS 分析同时监测造血细胞移植后的植入动态和白血病细胞

• 发表时间: 2011
• 作者: 渡辺恵理;中内祐介;渡辺信和;石田宏之;中内啓光
• 通讯作者: 中内啓光

Clinical diagnosis of graft failure and relapse of leukemia after HLA-mismatched hematopoietic cell transplantation by chimerism analysis using a spectral analyzer

光谱分析仪嵌合分析对HLA不匹配造血细胞移植后移植失败和白血病复发的临床诊断

• 发表时间: 2011
• 作者: Nobukazu Watanabe;Yusuke Nakauchi;Eri Watanabe;Masaya Kakuta;Koji Futamura;Motohiro Furuki;Hiromitsu Nakauchi
• 通讯作者: Hiromitsu Nakauchi

Engraftment kinetics of donor-derived lymphocytes early after reduced-intensity cord blood transplantation in adults.

成人低强度脐带血移植后早期供体来源淋巴细胞的植入动力学。

• 发表时间: 2010
• 作者: 簾藤紘子;佐々木純;磯部泰司;安田肇;今井英則;筒井深雪;押味和夫;杉本耕一;Naofumi Matsuno
• 通讯作者: Naofumi Matsuno

Flow cytometric analysis of donor-recipient chimerism using anti-HLA antibody after HLA-mismatched hematopoietic cell and organ transplantation.

HLA 不匹配的造血细胞和器官移植后，使用抗 HLA 抗体对供体-受体嵌合体进行流式细胞术分析。

• 发表时间: 2010
• 作者: Stephanie C.Napier;Satoshi Yamazaki;Nobukazu Watanabe;Naofumi Matsuno;Satoshi Takahashi;Akinobu Takaki;Hiromitsu Nakauchi
• 通讯作者: Hiromitsu Nakauchi