Liang Xiaoyan, Lu Lina, Chen Zongyou, Vickers Tim, Zhang Hong, Fung John J, Qian Shiguang
Department of Surgery, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania 15261, USA.
Transplantation. 2003 Aug 27;76(4):721-9. doi: 10.1097/01.TP.0000076470.35404.49.
The expression of costimulatory molecules on antigen-presenting cells is crucial in determining T-cell immune responses. We examined the effects of transduction with high-affinity antisense oligodeoxyribonucleotides (ODNs) designed to target the mRNA of CD80 or CD86 on the phenotype and function of dendritic cells (DCs).
DCs were propagated from C57BL/10 (B10; H2b) bone marrow cells in granulocyte macrophage-colony stimulating factor and interleukin (IL)-4, and transduced with anti-CD80 or anti-CD86 antisense ODNs (base-mismatched ODNs as controls). The effect of antisense ODN on phenotype was examined by flow cytometry. The allostimulatory function of DCs was assessed by mixed leukocyte reaction and cytotoxic activity in vitro, and the influence on allograft survival was assessed in vivo.
ODNs were effectively incorporated by DCs, which were enhanced by the presence of lipofectamine. Antisense ODNs targeting CD80 or CD86 mRNA specifically suppressed the expression of CD80 or CD86 in DCs and inhibited their capacity to elicit the proliferative responses, donor-specific cytotoxic T-lymphocyte activity in C3H (H2k) spleen T cells. This was associated with decreased IL-2, but elevated IL-4 production, and an increase in T-cell apoptosis. In contrast with the administration of control DCs into C3H recipients that exacerbated rejection of B10 cardiac allografts, injection of DCs transduced with anti-CD80 or CD86 antisense ODN significantly prolonged the survival of heart allografts.
Transduction with antisense ODN targeting CD80 or CD86mRNA is a feasible and effective approach to modify DCs that renders them tolerogenic by inducing T-cell hyporesponsiveness and apoptosis. This may lead to the development of new therapeutic strategies in transplantation.
抗原呈递细胞上共刺激分子的表达对于确定T细胞免疫反应至关重要。我们研究了用设计用于靶向CD80或CD86 mRNA的高亲和力反义寡脱氧核糖核苷酸（ODN）转导对树突状细胞（DC）表型和功能的影响。
从C57BL/10（B10；H2b）骨髓细胞在粒细胞巨噬细胞集落刺激因子和白细胞介素（IL）-4中培养DC，并用地高辛配基标记的抗CD80或抗CD86反义ODN（碱基错配的ODN作为对照）转导。通过流式细胞术检测反义ODN对表型的影响。通过混合淋巴细胞反应和体外细胞毒性活性评估DC的同种异体刺激功能，并在体内评估对同种异体移植存活的影响。
ODN被DC有效摄取，lipofectamine的存在增强了摄取。靶向CD80或CD86 mRNA的反义ODN特异性抑制DC中CD80或CD86的表达，并抑制其引发C3H（H2k）脾T细胞增殖反应、供体特异性细胞毒性T淋巴细胞活性的能力。这与IL-2产生减少但IL-4产生增加以及T细胞凋亡增加有关。与将对照DC注入C3H受体加剧B10心脏同种异体移植排斥反应相反，注射用抗CD80或抗CD86反义ODN转导的DC显著延长了心脏同种异体移植的存活时间。
用靶向CD80或CD86 mRNA的反义ODN转导是一种可行且有效的修饰DC的方法，可通过诱导T细胞低反应性和凋亡使其具有耐受性。这可能会导致移植新治疗策略的发展。
