Activation of Antigen-Specific Cytotoxic T Lymphocytes by β2-Microglobulin or TAP1 Gene Disruption and the Introduction of Recipient-Matched MHC Class I Gene in Allogeneic Embryonic Stem Cell-Derived Dendritic Cells
Activation of Antigen-Specific Cytotoxic T Lymphocytes by β2-Microglobulin or TAP1 Gene Disruption and the Introduction of Recipient-Matched MHC Class I Gene in Allogeneic Embryonic Stem Cell-Derived Dendritic Cells
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DOI：
10.4049/jimmunol.181.9.6635
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发表时间：
2008-11-01
影响因子：
4.4
通讯作者：
Senju, Satoru
中科院分区：
文献类型：
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作者：
Matsunaga, Yusuke;Fukuma, Daiki;Senju, Satoru
树突状细胞（DC）的遗传修饰方法是基于胚胎干细胞（ES）体外分化为DC（ES-DC）而建立的。与患者基因相同的人类ES细胞的不可用性将是该技术未来临床应用的一个问题。本研究试图建立一个战略，以克服这一问题。在129（H-2（B））衍生的ES细胞中破坏TAPI或β（2）-微球蛋白（β（2）m）基因，然后引入H-2 K（d）或β（2）m-连接形式的K-d（β 2 m-K-d）的表达载体，从而产生两种类型的基因工程ES-DC，TAPI（-/-）/K-d ES-DC和β M-2（-/-）/β（2）m-K-d ES-DC。如预期的，两个转染子ES-DC表达K-d，但不表达固有H-2（B）单倍型衍生的MHC I类。β（2）m（-/-）/β（2）m-K-d和TAP 1（-/-）/K-d ES-DC不被预活化的H-2（B）-反应性CTL识别，并且在体外或体内不引发H-2（B）-反应性CTL。当转移到BALB/c小鼠中时，与β（2）m（+/-）/β（2）m-K-d ES-DC和TAP 1（+/+）/K-d ES-DC相比，β（2）m（-/-）/β（2）m-K-d ES-DC和TAP 1（-/-）/K-d ES-DC具有存活优势。负载K-d限制性RSV-M2衍生肽的ES-DC在注射到BALB/c小鼠后可引发表位特异性CTL，不考虑细胞表面内源性H-2（B）单倍型编码的MHC I类分子的表达，β（2）m（-/-）/β（2）m-K-dES-DC在诱导针对RSV M2蛋白表达肿瘤细胞的免疫方面比β（2）m（+/-）/β（2）m-K-dES-DC显著更有效。d ES-DC。因此，β（2）m或TAP基因的修饰可能是解决人类ES或诱导多能干细胞与待治疗的受体之间的HLA I类等位基因错配问题的有效策略。免疫学杂志，2008，181：6635-6643.
A method for the genetic modification of dendritic cells (DC) was previously established based on the in vitro differentiation of embryonic stem (ES) cells to DC (ES-DC). The unavailability of human ES cells genetically identical to the patients will be a problem in the future clinical application of this technology. This study attempted to establish a strategy to overcome this issue. The TAPI or beta(2)-microglobulin (beta(2)m) gene was disrupted in 129 (H-2(b))-derived ES cells and then expression vectors for the H-2K(d) or beta(2)m-linked form of K-d (beta 2m-K-d) were introduced, thus resulting in two types of genetically engineered ES-DC, TAP1(-/-)/K-d ES-DC and beta M-2(-/-)/beta(2)m-K-d ES-DC. As intended, both of the transfectant ES-DC expressed K-d but not the intrinsic H-2(b) haplotype-derived MHC class I. beta(2)m(-/-)/beta(2)m-K-d and TAP1(-/-)/K-d ES-DC were not recognized by pre-activated H-2(b)-reactive CTL and did not prime H-2(b) reactive CTL in vitro or in vivo. beta(2)m(-/-)/beta(2)m-K-d ES-DC and TAP1(-/-)/K-d ES-DC had a survival advantage in comparison to beta(2)m(+/-)/beta(2)m-K-d ES-DC and TAP1(+/+)/K-d ES-DC, when transferred into BALB/c mice. K-d-restricted RSV-M2-derived peptide-loaded ES-DC could prime the epitope-specific CTL upon injection into the BALB/c mice, irrespective of the cell surface expression of intrinsic H-2(b) haplotype-encoded MHC class I.beta(2)m(-/-)/beta(2)m-K-d ES-DC were significantly more efficient in eliciting immunity against RSV M2 protein-expressing tumor cells than beta(2)m(+/-)/beta(2)m-K-d ES-DC. The modification of the beta(2)m or TAP gene may therefore be an effective strategy to resolve the problem of HLA class I allele mismatch between human ES or induced pluripotent stem cells and the recipients to be treated. The Journal of Immunology, 2008, 181: 6635-6643.