In vivo correction of ZAP-70 immunodeficiency by intrathymic gene transfer
In vivo correction of ZAP-70 immunodeficiency by intrathymic gene transfer
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DOI：
10.1172/jci23966
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发表时间：
2005-08-01
影响因子：
15.9
通讯作者：
Klatzmann, D
中科院分区：
文献类型：
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作者：
Adjali, O;Marodon, G;Klatzmann, D
SCID患者已经通过体外基因校正干细胞成功治疗。然而，尽管这种治疗已被证明有效，但它也有特定的风险和困难。我们假设这些缺陷可以通过胸腺T淋巴祖细胞的原位基因校正来克服。事实上，在体内胸腺内转移一种为转导的胸腺原细胞提供选择优势的基因，应该导致功能性T淋巴细胞后代的产生，从而允许长期的免疫重建。我们在小鼠zap -70缺陷SCID模型中评估了这种方法的可行性。将一种T细胞特异性表达ZAP-70的慢病毒载体注射到成年ZAP-70（-/-）小鼠胸腺中，无需事先调节。这导致成熟的tcr - α - β(+)胸腺细胞长期分化，表明该载体已整合到祖细胞中。此外，外周表达zap -70的T细胞表现出部分多样化的受体库，并在体外和体内对同种异体抗原有反应。在幼龄ZAP-70（-/-）小鼠中，胸腺相应增大，胸腺原细胞处于周期的比例更高，治疗效果得到改善。因此，胸腺内注射慢病毒载体可能是一种简化的、潜在的更安全的替代体外基因修饰造血干细胞移植的T细胞免疫缺陷基因治疗方法。
SCID patients have been successfully treated by administration of ex vivo gene-corrected stem cells. However, despite its proven efficacy, such treatment carries specific risks and difficulties. We hypothesized that some of these drawbacks may be overcome by in situ gene correction of T lymphoid progenitors in the thymus. Indeed, in vivo intrathymic transfer of a gene that provides a selective advantage for transduced prothymocytes should result in the generation of functional T lymphocyte progeny, allowing long-term immune reconstitution. We assessed the feasibility of this approach in a murine model of ZAP-70-deficient SCID. A T cell-specific ZAP-70-expressing lentiviral vector was injected into thymi of adult ZAP-70(-/-) mice without prior conditioning. This resulted in the long-term differentiation of mature TCR-alpha beta(+) thymocytes, indicating that the vector had integrated into progenitor cells. Moreover, peripheral ZAP-70-expressing T cells demonstrated a partially diversified receptor repertoire and were responsive to alloantigens in vitro and in vivo. Improved treatment efficacy was achieved in infant ZAP-70(-/-) mice, in which the thymus is proportionately larger and a higher percentage of prothymocytes are in cycle. Thus, intrathymic injection of a lentiviral vector could represent a simplified and potentially safer alternative to ex vivo gene-modified hematopoietic stem cell transplantation for gene therapy of T cell immunodeficiencies.