...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Protein Expression and Purification >Refinement in the production and purification of recombinant HCMV IE1-pp65 protein for the generation of epitope-specific T cell immunity
【24h】

Refinement in the production and purification of recombinant HCMV IE1-pp65 protein for the generation of epitope-specific T cell immunity

机译：精细生产和纯化重组HCMV IE1-pp65蛋白以产生表位特异性T细胞免疫

获取原文
获取原文并翻译 | 示例
   

获取外文期刊封面封底 >>

       
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Human cytomegalovirus (HCMV) remains one of the most common opportunistic infections causing disease following stem cell transplantation, despite the availability of anti-viral therapies. Adoptive immunotherapy has the potential to further aid in counteracting chronic viral reactivation and subsequent disease by restoring viral immunity through the transfer of virus-specific T cells from transplant donors to their recipients. Our study refines the production and purification of a recombinant HCMV protein containing two of the most immunodominant antigens (IE1 and pp65) for the generation of polyclonal HCMV-specific T cells. In doing so, a 6 x His-tagged IE1-pp65 protein was generated using a serum-free baculovirus/insect cell expression system and soluble IE1-pp65 protein was subsequently purified using Ni-NTA affinity chromatography under stringent conditions to obtain a highly pure product. The ability of the recombinant IE1-pp65 protein to elicit a functional T cell mediated immune response was demonstrated by the vigorous reactivation and expansion of HLA-A2-restricted pp65(495-503)-specific CD8(+) T cells. This recombinant IE1-pp65 protein can potentially generate a multitude of HLA-restricted HCMV-specific T cells, providing a better alternative to using costly overlapping peptides or HCMV lysates for expansion of T cells for use in adoptive immunotherapy strategies. (C) 2008 Elsevier Inc. All rights reserved.
机译：尽管有抗病毒疗法，人类巨细胞病毒（HCMV）仍然是引起干细胞移植后疾病的最常见机会性感染之一。过继免疫疗法有潜力通过将病毒特异性T细胞从移植供体转移到受体来恢复病毒免疫力，从而进一步帮助抵抗慢性病毒再激活和随后的疾病。我们的研究改进了包含两种最主要免疫抗原（IE1和pp65）的重组HCMV蛋白的生产和纯化，以产生多克隆HCMV特异性T细胞。为此，使用无血清杆状病毒/昆虫细胞表达系统生成了6 x His标记的IE1-pp65蛋白，随后在严格条件下使用Ni-NTA亲和色谱法纯化了可溶性IE1-pp65蛋白，从而获得了高纯度的产品。重组IE1-pp65蛋白引发功能性T细胞介导的免疫反应的能力已通过HLA-A2限制性pp65（495-503）特异性CD8（+）T细胞的强烈活化和扩增得到证实。这种重组IE1-pp65蛋白可以潜在地产生多种HLA限制的HCMV特异性T细胞，为使用昂贵的重叠肽或HCMV裂解物扩增T细胞用于过继免疫疗法提供了更好的选择。 （C）2008 Elsevier Inc.保留所有权利。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号