Otsu Makoto, Steinberg Marcos, Ferrand Christophe, Merida Peggy, Rebouissou Cosette, Tiberghien Pierre, Taylor Naomi, Candotti Fabio, Noraz Nelly
Genetics and Molecular Biology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA.
Blood. 2002 Aug 15;100(4):1248-56. doi: 10.1182/blood-2002-01-0247.
Mutations in the ZAP-70 protein tyrosine kinase gene result in a severe combined immunodeficiency (SCID) characterized by a selective inability to produce CD8(+) T cells and a signal transduction defect in peripheral CD4(+) cells. Transplantation of genetically modified hematopoietic progenitor cells that express the wild-type ZAP-70 gene may provide significant benefit to some of these infants. The feasibility of stem cell gene correction for human ZAP-70 deficiency was assessed using a ZAP-70 knock-out model. ZAP-70-deficient murine bone marrow progenitor cells were transduced with a retroviral vector expressing the human ZAP-70 gene. Engraftment of these cells in irradiated ZAP-70-deficient animals resulted in the development of mature CD4(+) and CD8(+) T cells. In marked contrast, both populations were absent in ZAP-70(-/-) mice undergoing transplantation with bone marrow progenitor cells transduced with a control vector. Importantly, ZAP-70-reconstituted T cells proliferated in response to T-cell receptor stimulation. Moreover, these ZAP-70-expressing T cells demonstrated a diverse T-cell receptor repertoire as monitored by the relative usage of each T-cell receptor beta chain hypervariable region subfamily. The presence of ZAP-70 in B cells did not affect either lipopolysaccharide- or lipopolysaccharide/interleukin-4-mediated immunoglobulin isotype switching. Altogether, these data indicate that retroviral-mediated gene transfer of the ZAP-70 gene may prove to have a therapeutic benefit for patients with ZAP-70-SCID.
ZAP-70蛋白酪氨酸激酶基因突变会导致严重联合免疫缺陷（SCID），其特征是选择性无法产生CD8(+) T细胞以及外周CD4(+)细胞存在信号转导缺陷。移植表达野生型ZAP-70基因的基因修饰造血祖细胞可能会给部分此类婴儿带来显著益处。利用ZAP-70基因敲除模型评估了对人类ZAP-70缺陷进行干细胞基因校正的可行性。用表达人类ZAP-70基因的逆转录病毒载体转导ZAP-70缺陷的小鼠骨髓祖细胞。将这些细胞移植到经辐照的ZAP-70缺陷动物体内后，成熟的CD4(+)和CD8(+) T细胞得以发育。与之形成鲜明对比的是，在用对照载体转导的骨髓祖细胞进行移植的ZAP-70(-/-)小鼠中，这两种细胞群均不存在。重要的是，ZAP-70重构的T细胞会因T细胞受体刺激而增殖。此外，通过监测每个T细胞受体β链高变区亚家族的相对使用情况发现，这些表达ZAP-70的T细胞表现出多样化的T细胞受体库。B细胞中ZAP-70的存在并不影响脂多糖或脂多糖/白细胞介素-4介导的免疫球蛋白同种型转换。总之，这些数据表明，ZAP-70基因的逆转录病毒介导基因转移可能对ZAP-70-SCID患者具有治疗益处。
