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经RNA电穿孔的CD40激活的B细胞诱导针对HepG2细胞的功能性T细胞反应。

RNA-electroporated CD40-activated B cells induce functional T-cell responses against HepG2 cells.

作者信息

Shen S N, Xu Z, Qian X P, Ding Y T, Yu L X, Liu B R

机构信息

Department of Oncology, The Affiliated Drum Tower Hospital, Medical School of Naniing University and Clinical Cancer Institute of Nanjing University, Naniing, China.

出版信息

Eur J Cancer Care (Engl). 2008 Jul;17(4):404-11. doi: 10.1111/j.1365-2354.2007.00841.x.

Abstract

Hepatocellular carcinoma (HCC) is one of the incurable tumours in the world. Cell-based immunotherapy, in which antigen-loaded antigen-presenting cells (APCs) are able to elicit T cell responses, has become an alternative treatment for liver cancer. Here, we used HepG2 cells' total RNA-electroporated CD40 ligand-activated B (CD40-B) cells as alternative APC for induction of specific CD8+ T-cell responses. The antigen-presenting ability of CD40-B cells was determined by phenotypic analysis, showing a polyclonal, strongly activated B-cell population with high expression of co-stimulatory molecules. To demonstrate the ability of total RNA extracted from HepG2 cells electroporated CD40-B cells to induce CD8+ T-cell responses, these RNA-loaded cells were co-cultured with autologous peripheral blood mononuclear cells for 7 days followed by analysis of T-cell antigen specificity. These experiments showed that CD40-B cells electroporated with HepG2 cells' total RNA are capable of activating antigen-specific interferon-gamma-producing CD8+ T cells, and these T cells activated by CD40-B cells show a killing effect on HepG2 cells. These findings demonstrated that the carcinoma cell derived total RNA-electroporated CD40-B cells could be used as alternative APC for the induction of antigen-specific CD8+ T-cell responses, which might be used in HCC immunotherapy.

摘要

肝细胞癌(HCC)是世界上难以治愈的肿瘤之一。基于细胞的免疫疗法,即负载抗原的抗原呈递细胞(APC)能够引发T细胞反应,已成为肝癌的一种替代治疗方法。在此,我们使用经HepG2细胞总RNA电穿孔的CD40配体激活的B细胞(CD40 - B细胞)作为替代抗原呈递细胞来诱导特异性CD8 + T细胞反应。通过表型分析确定CD40 - B细胞的抗原呈递能力,结果显示其为具有共刺激分子高表达的多克隆、强烈激活的B细胞群体。为了证明从经HepG2细胞电穿孔的CD40 - B细胞中提取的总RNA诱导CD8 + T细胞反应的能力,将这些负载RNA的细胞与自体外周血单个核细胞共培养7天,随后分析T细胞抗原特异性。这些实验表明,用HepG2细胞总RNA电穿孔的CD40 - B细胞能够激活产生抗原特异性干扰素 - γ的CD8 + T细胞,并且这些由CD40 - B细胞激活的T细胞对HepG2细胞具有杀伤作用。这些发现表明,癌细胞来源的总RNA电穿孔的CD40 - B细胞可作为诱导抗原特异性CD8 + T细胞反应的替代抗原呈递细胞,这可能用于HCC免疫治疗。

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