Stepkowski S M, Tu Y, Condon T P, Bennett C F
Department of Surgery, University of Texas Medical School at Houston 77030.
J Immunol. 1994 Dec 1;153(11):5336-46.
Intercellular adhesion molecule-1 (ICAM-1) binds circulating leukocytes through interactions with beta 2 integrins, LFA-1, and macrophage Ag-1. The phosphorothioate antisense oligodeoxynucleotide, IP-3082, specific for ICAM-1 mRNA inhibited ICAM-1, but not vascular cell adhesion molecule-1, mRNA induction and expression of ICAM-1 molecules by mouse endothelioma cells. Scrambled control oligonucleotides were ineffective. Untreated C3H (H-2k) mice rejected C57BL/10 (H-2b) heart allografts with a mean survival time of 7.7 +/- 1.4 days. Administration i.v. of IP-3082 by a 7-day osmotic pump prolonged the survival of heart allografts in a dose-dependent fashion: 1.25 mg/kg, to 11 +/- 0 days; 2.5 mg/kg, to 12 +/- 2.7 days; 5 mg/kg, to 14.1 +/- 2.7 days; and 10 mg/kg, to 15.3 +/- 5.8 days (all p < 0.01). Control IP-1082 (10 mg/kg) was ineffective (7 +/- 0.8 days). Although 7-day anti-LFA-1 mAb (50 micrograms/day; i.p.) prolonged allograft survival to 14.1 +/- 2.7 days, the addition of IP-3082 (5.0 mg/kg x 7 days) induced donor-specific transplantation tolerance (> 150 days). Furthermore, IP-3082 (5 mg/kg x 7 days) acted synergistically with antilymphocyte serum, rapamycin, and brequinar, but not cyclosporin A: a single antilymphocyte serum (0.2 ml) i.p. injection alone prolonged graft survival to 10 +/- 0.5 days (p < 0.01) and in combination with IP-3082 (5 mg/kg), to 32.2 +/- 8.3 days (p < 0.001); rapamycin (0.1 mg/kg x 7 days; i.v.) alone prolonged survival to 13 +/- 7.5 days (p < 0.01), and with IP-3082, to 32.4 +/- 8.9 days (p < 0.03); brequinar (0.5 mg/kg x 7 days; oral gavage) alone to 12 +/- 2.4 days (p < 0.05), and with IP-3082 (5 mg/kg), to 38.8 +/- 30.2 days (p < 0.01); in contrast, cyclosporin A (5 mg/kg x 7 days; i.v.) alone produced graft survival of 9.8 +/- 1.3 days (p < 0.1 and in combination with IP-3082 (5 mg/kg), produced survival of 7.8 +/- 3.5 days (NS). Thus, antisense oligonucleotides may proffer a selective gene-targeted immunosuppressive therapy for organ transplantation.
细胞间黏附分子-1（ICAM-1）通过与β2整合素、淋巴细胞功能相关抗原-1（LFA-1）和巨噬细胞抗原-1相互作用，结合循环中的白细胞。针对ICAM-1 mRNA的硫代磷酸酯反义寡脱氧核苷酸IP-3082可抑制ICAM-1，但不抑制血管细胞黏附分子-1的mRNA诱导以及小鼠内皮瘤细胞中ICAM-1分子的表达。乱序对照寡核苷酸则无效。未处理的C3H（H-2k）小鼠排斥C57BL/10（H-2b）心脏同种异体移植物，平均存活时间为7.7±1.4天。通过7天渗透泵静脉注射IP-3082可使心脏同种异体移植物的存活时间以剂量依赖方式延长：1.25mg/kg时，延长至11±0天；2.5mg/kg时，延长至12±2.7天；5mg/kg时，延长至14.1±2.7天；10mg/kg时，延长至15.3±5.8天（所有p<0.01）。对照IP-1082（10mg/kg）无效（7±0.8天）。尽管7天抗LFA-1单克隆抗体（50μg/天；腹腔注射）可使同种异体移植物存活时间延长至14.1±2.7天，但添加IP-3082（5.0mg/kg×7天）可诱导供体特异性移植耐受（>150天）。此外，IP-3082（5mg/kg×7天）与抗淋巴细胞血清、雷帕霉素和布喹那具有协同作用，但与环孢素A无协同作用：单独腹腔注射一次抗淋巴细胞血清（0.2ml）可使移植物存活时间延长至10±0.5天（p<0.01），与IP-3082（5mg/kg）联合使用时，延长至32.2±8.3天（p<0.001）；雷帕霉素（0.1mg/kg×7天；静脉注射）单独使用时可使存活时间延长至13±7.5天（p<0.01），与IP-3082联合使用时，延长至32.4±8.9天（p<0.03）；布喹那（0.5mg/kg×7天；灌胃）单独使用时可使存活时间延长至12±2.4天（p<0.05），与IP-3082（5mg/kg）联合使用时，延长至38.8±30.2天（p<0.01）；相比之下，环孢素A（5mg/kg×7天；静脉注射）单独使用时移植物存活时间为9.8±1.3天（p<0.1），与IP-3082（5mg/kg）联合使用时，存活时间为7.8±3.5天（无显著性差异）。因此，反义寡核苷酸可能为器官移植提供一种选择性基因靶向免疫抑制疗法。
