CD4-TARGETED EXPRESSION OF HIV PROTECTIVE GENES

HIV疾病的保护性细胞内基因治疗需要靶向 将候选基因表达到作为主要细胞的细胞中， 艾滋病毒的靶点，即巨噬细胞和辅助性T细胞，两者都是 表达细胞表面CD 4糖蛋白。 我们最近发现 调节元件参与了积极和消极的 CD 4基因的转录调控。 我们将定义最小CD 4 可用于在T辅助细胞中获得表达的调节序列 细胞和各种单核细胞/巨噬细胞谱系细胞。 我们还将寻求 为了通过测试品种获得高水平的谱系限制表达， 其中一些正在开发中， 在纳贝尔博士的实验室里 我们将利用这些信息来开发 用于逆转录病毒转导和用于颗粒介导递送的载体 这将指导治疗候选物的表达， 导入骨髓干细胞后纠正造血谱系 细胞和祖细胞。 我们将与Kohn博士一起测试和优化 这些载体在鼠骨髓重建系统中。 如果 如果在小鼠系统中的实验成功，则将使用载体 将HIV转显性基因植入人类骨髓细胞， 血清阳性个体。

Protective intracellular gene therapy for HIV disease requires targeting expression of candidate genes to the cells that serve as the primary targets of HIV, namely macrophages and T helper cells, both of which express cell surface CD4 glycoprotein. We have recently identified regulatory elements involved in both positive and negative transcriptional regulation of the CD4 gene. We will define minimal CD4 regulatory sequences that can be used to attain expression in T helper cells and various monocyte/macrophage lineage cells. We will also seek to attain high level lineage-restricted expression by testing a variety of combinations of regulatory elements, some of which are being developed in Dr. Nabel's laboratory. We will use this information to develop vectors for retroviral transduction and for particle-mediated delivery that will direct expression of therapeutic candidates exclusively in the correct hematopoietic lineages after introduction into bone marrow stem cells and progenitors. We will work with Dr. Kohn to test and optimize these vectors in a murine bone marrow reconstitution system. If experiments in murine systems are successful, the vectors will be used to transduce HIV transdominant genes into human bone marrow cells in seropositive individuals.