Nair S K, Snyder D, Gilboa E
Department of Surgery, Duke University Medical Center, Durham, NC 27710 USA.
J Immunol. 1996 Mar 1;156(5):1772-80.
Treatment of RMA and EL4 cells or freshly isolated splenocytes with antisense (AS) oligonucleotides directed against the TAP-2 gene recreates the phenotype seen in cells that are genetically deficient in TAP function. Cells incubated with AS oligonucleotides exhibit reduced MHC class I expression on the cell surface, which can be increased by incubating the oligonucleotide-treated cells at 28 degrees C or by adding MHC haplotype-matched peptides to the culture medium. RMA cells or splenocytes treated with AS oligonucleotides and incubated with peptide were highly effective in generating primary CTL responses in vitro. The bulk of the AS oligonucleotide-responsive and CTL-inducing cells resided in the adherent fraction of splenocytes. Moreover, TAP-2 AS oligonucleotide-treated adherent splenocytes pulsed with OVA peptide elicited potent OVA-specific CTL responses in vivo and provided effective protection from challenge with tumor cells expressing the corresponding Ag. AS oligonucleotide technology provides a simple approach to develop broadly applicable methods for generating potent APC to study TAP function in normal cells and to identify other gene products involved in MHC class I presentation.
用针对TAP-2基因的反义（AS）寡核苷酸处理RMA和EL4细胞或新鲜分离的脾细胞，可重现TAP功能基因缺陷细胞中所见的表型。用AS寡核苷酸孵育的细胞在细胞表面表现出MHC I类表达降低，将经寡核苷酸处理的细胞在28℃孵育或向培养基中添加MHC单倍型匹配的肽可增加这种表达。用AS寡核苷酸处理并与肽一起孵育的RMA细胞或脾细胞在体外产生初级CTL反应方面非常有效。大部分对AS寡核苷酸有反应并诱导CTL的细胞存在于脾细胞的贴壁部分。此外，用OVA肽脉冲处理的经TAP-2 AS寡核苷酸处理的贴壁脾细胞在体内引发了强大的OVA特异性CTL反应，并为抵御表达相应抗原的肿瘤细胞攻击提供了有效保护。AS寡核苷酸技术提供了一种简单的方法，可开发广泛适用的方法来生成有效的抗原呈递细胞，以研究正常细胞中的TAP功能，并鉴定参与MHC I类呈递的其他基因产物。
