Salgar S K, Yang D, Ruiz P, Miller J, Tzakis A G
Department of Surgery, University of Miami, Miami, FL 33136, USA.
Transplant Proc. 2004 Mar;36(2):397-8. doi: 10.1016/j.transproceed.2003.12.009.
In this study, a novel gene therapy approach to prolong allograft survival was designed. Autologous (syngeneic) hematopoietic stem cell-enriched bone marrow cells (HSC; lin(-)) engineered with the vIL-10 gene (vIL-10-HSC) were injected (4 to 6 x 10(6) cells, i.v.) into lethally (9.5 Gy) or sublethally (4 Gy) irradiated CBA/J mice 6 weeks prior to allogeneic heart (C57BL/6) transplantation (Tx). Cardiac allograft survival was significantly (P <.004) prolonged in lethally (71 +/- 40 days) and sublethally (114 +/- 15 days) irradiated mice that received vIL-10-HSC compared to controls that received no HSC (11 +/- 1 days), unengineered HSC, or vector-DNA-engineered HSC (12 to 16 days). Tolerant graft histopathology demonstrated mild arteritis/venulitis (grade 0.7) and rejection (grade 1.0). Intragraft expression of costimulatory molecules (B7.1, B7.2), cytokines (IL-2, IL-4, mIL-10, IFN-gamma), and iNOS molecules were markedly lower in tolerant grafts that survived for >100 days; recipient T lymphocytes demonstrated hyporeactivity to donor and third-party antigens in mixed lymphocyte cultures. These findings have important implications and potential therapeutic applications in transplantation and autoimmune diseases.
在本研究中，设计了一种延长同种异体移植物存活时间的新型基因治疗方法。将经vIL-10基因工程改造的自体（同基因）富含造血干细胞的骨髓细胞（HSC；lin(-)）（4至6×10⁶个细胞，静脉注射）在同种异体心脏（C57BL/6）移植（Tx）前6周注入致死剂量（9.5 Gy）或亚致死剂量（4 Gy）照射的CBA/J小鼠体内。与未接受HSC的对照组（11±1天）、未进行基因工程改造的HSC组或载体-DNA基因工程改造的HSC组（12至16天）相比，接受vIL-10-HSC的致死照射（71±40天）和亚致死照射（114±15天）小鼠的心脏同种异体移植物存活时间显著延长（P<.004）。耐受移植物的组织病理学表现为轻度动脉炎/静脉炎（0.7级）和排斥反应（1.0级）。在存活超过100天的耐受移植物中，共刺激分子（B7.1、B7.2）、细胞因子（IL-2、IL-4、mIL-10、IFN-γ）和iNOS分子的移植物内表达明显较低；在混合淋巴细胞培养中，受体T淋巴细胞对供体和第三方抗原表现出低反应性。这些发现对移植和自身免疫性疾病具有重要意义和潜在的治疗应用价值。
