Gene transfer in vivo with DNA-liposome complexes: lack of autoimmunity and gonadal localization.
Gene transfer in vivo with DNA-liposome complexes: lack of autoimmunity and gonadal localization.
复制标题
DNA-脂质体复合物体内基因转移：缺乏自身免疫和性腺定位。
作者：
E. Nabel;D. Gordon;Zhi;Ling Xu;H. San;G. Plautz;Bei;Xiang Gao;Leaf Huang;G. Nabel
直接将基因转移到局部动脉节段可以通过DNA-脂质体复合体在体内进行。这项技术有望用于治疗人类疾病，包括恶性肿瘤和心血管疾病。我们以前已经在小鼠体内表征了这种治疗形式的潜在毒性(Stewart等人，1992)。在这份报告中，我们研究了与外源重组基因长期表达相关的两个问题：(I)主要器官的自身免疫性损伤的可能性和(Ii)性腺组织中的DNA定位。通过体内直接基因转移诱导小鼠对重组小鼠主要组织相容性(MHC)基因的免疫应答后，对同种异体主要组织相容性(MHC)基因转移的自身免疫和毒性进行了评估。对脑、心、肺、肝、肾、脾和骨骼肌的组织学检查显示，临床上没有明显的免疫病理学或器官损害。在猪和兔体内，分析了DNA脂质体基因传递的毒性。在引入编码几种不同基因产物的质粒后，没有观察到组织病理学，并且DNA脂质体注射后的血清分析显示，血清生化参数没有异常。通过聚合酶链式反应评价重组DNA转移到动物睾丸和卵巢的可能性。虽然在已转染者的动脉部位持续观察到重组质粒的存在，但在未转染者的动脉部位并未观察到，偶尔在肺、肾、脾和肝脏中也能检测到，但在睾丸或卵巢中未检测到质粒DNA。这些研究表明，在主要器官通过脂质体直接基因转移后，对重组DNA的摄取与自身免疫、毒性或性腺定位无关。
Direct gene transfer into localized arterial segments can be performed in vivo by transfection with DNA-liposome complexes. This technique holds promise for the treatment of human diseases, including malignancy and cardiovascular disorders. We have previously characterized the potential toxicity of this form of treatment in mice in vivo (Stewart et al., 1992). In this report, we examined two issues relevant to long-term expression of foreign recombinant genes: (i) the potential for autoimmune damage to major organs and (ii) DNA localization in gonadal tissue. Autoimmunity and toxicity of allogeneic major histocompatibility (MHC) gene transfer was assessed in mice after induction of an immune response to a recombinant murine class I MHC gene by direct gene transfer in vivo. Histological examination of brain, heart, lung, liver, kidney, spleen, and skeletal muscle revealed no clinically significant immunopathology or organ damage. The toxicity of gene delivery by DNA liposomes was also analyzed in pigs and rabbits in vivo. No histopathology was observed following the introduction of plasmids encoding several different gene products, and analysis of serum following DNA liposome delivery revealed no abnormalities of serum biochemical parameters. The potential for transfer of recombinant DNA into testes and ovary in animals was evaluated by the polymerase chain reaction. Although evidence of recombinant plasmid was consistently observed in transfected, but not untransfected, arterial sites and occasionally in lung, kidney, spleen, and liver, no plasmid DNA was detected in testes or ovary. These studies suggest that uptake of recombinant DNA following direct gene transfer by liposomal transfection in major organs is not associated with autoimmunity, toxicity, or gonadal localization.(ABSTRACT TRUNCATED AT 250 WORDS)