Valere T, Bohl D, Klatzmann D, Danos O, Sonigo P, Heard J M
Institut Cochin de Génétique Moléculaire (ICGM-CNRS UPR415), Paris, France.
Gene Ther. 1995 May;2(3):197-202.
Somatic transgenesis can be used to confer endogenous production of proteins with therapeutic properties. One such product, recombinant soluble human CD4 (sCD4), has been shown to be an efficient inhibitor of human immunodeficiency virus 1 (HIV-1) in vitro, but its too short half-life in vivo has impaired long-term clinical trials in AIDS patients. Using a retroviral vector, we introduced the cDNA of sCD4 into primary mouse fibroblasts. The cells were enclosed in a lattice of collagen and synthetic fibers coated with basic fibroblast growth factor, and implanted in the peritoneal cavity of syngeneic mice. Implantation of such sCD4-secreting organoids into cyclosporin A-treated C3H mice elicited a strong antibody response against sCD4. Implantation of sCD4-secreting organoids into immunotolerant mice (transgenic for transmembrane human CD4) resulted in continuous sCD4 production, detected during 60 days in animal sera. The serum levels obtained were significant, but too limited as yet for anti-HIV purposes. Nevertheless, this model may be of interest in various fields, as it provides the first demonstration that one potentially therapeutic protein, despite its half-life of a few hours, could remain present in vivo 2 months after a single somatic transgenesis.
体细胞转基因可用于赋予具有治疗特性的蛋白质内源性产生。一种这样的产物，重组可溶性人CD4（sCD4），已被证明在体外是人类免疫缺陷病毒1（HIV-1）的有效抑制剂，但其在体内的半衰期过短影响了在艾滋病患者中的长期临床试验。使用逆转录病毒载体，我们将sCD4的cDNA导入原代小鼠成纤维细胞。将细胞包裹在涂有碱性成纤维细胞生长因子的胶原蛋白和合成纤维晶格中，并植入同基因小鼠的腹腔。将这种分泌sCD4的类器官植入环孢素A处理的C3H小鼠中引发了针对sCD4的强烈抗体反应。将分泌sCD4的类器官植入免疫耐受小鼠（跨膜人CD4转基因）中导致持续产生sCD4，在动物血清中60天内均可检测到。获得的血清水平是显著的，但目前对于抗HIV目的来说还太有限。然而，这个模型可能在各个领域都有意义，因为它首次证明了一种潜在的治疗性蛋白质，尽管其半衰期只有几个小时，但在单次体细胞转基因后两个月仍可在体内存在。
