LIPIDATED ANTIBODIES FOR INTRACELLULAR THERAPY OF HIV

有效的HIV-1治疗是一个复杂的问题，可能需要至少 两种不同的方法。我们建议解决阻塞问题 病毒复制与免疫构建的启动有关。 已经开发了一种共价连接脂胺的新技术， 甘氨酰双十八烷基酰胺，多克隆或 克隆抗体碳水化合物残基位于fc上 从高变区去除的抗体的一部分，和 因此不干扰抗体结合。的存在 脂胺似乎促进抗体通过血浆 活细胞的膜而不丧失表位特异性。这 通过直接染色细胞内 细胞骨架蛋白这些抗体还具有以下能力： 中和细胞内蛋白质功能。初步数据表明 单独地，脂化的抗Tat，抗p24/25 gag，和抗逆转录病毒， 转录酶抗体可以显著减少HIV病毒的复制， 111 B和野生型分离株在感染的SupT 1细胞和潜伏 感染J1.1和OM 10.1细胞。另外，我们克隆了一个淋巴细胞 对CD 4+细胞特异的感受态生长因子。淋巴因子（LCF） 诱导IL-2 R的表达并使细胞对IL-2产生应答 刺激.前两个目标的重点是体外优化 用于脂化抗体阻断HIV-1复制的条件，和 优化LCF的CD 4+淋巴细胞生长潜力。这包括 筛选一组脂化抗HIV-1病毒蛋白抗体， 他们阻断HIV-1复制的能力，单独或在不同的 组合。基于体外数据，我们的第三个目标是应用这些 两种新的方法直接评估抗体阻断 和HIV-1的SCID小鼠动物模型中的免疫构建。

Effective HIV-1 therapy is a complex problem which may require at least two different approaches. We propose to address the issues of blocking viral replication in conjunction with initiation of immunoreconstitution. A novel technology has been developed which covalently links a lipoamine, glycyldioctadecylamide, to the carbohydrate moieties of polyclonal or monoclonal antibodies. The carbohydrate residues are located on the fc portion of the antibody, removed from the hypervariable region, and consequently do not interfere with antibody binding. The presence of the lipoamines appear to promote passage of the antibody through the plasma membrane of living cells without loss of epitope specificity. This has been demonstrated visually by direct staining of intracellular cytoskeletal proteins. These antibodies also contain the capability of neutralizing intracellular protein function. Preliminary data indicates that individually, lipidated anti-Tat, anti-p24/25 gag, and anti-reverse transcriptase antibodies can markedly reduced viral replication of HIV- 1IllB and wild-type isolates in infected SupT1 cells and latently infected J1.1 and OM 10.1 cells. In addition, we have cloned a lymphocyte competence growth factor specific for CD4+ cells. This lymphokine (LCF) induces the expression of IL-2R and renders the cells responsive to IL-2 stimulation. The focus of the first two aims is to optimize in vitro conditions for lipidated antibody blocking of HIV-l replication, and optimize the CD4 + lymphocyte growth potential of LCF. This includes screening a panel of lipidated anti-HIV-1 viral protein antibodies for their ability to block HIV-1 replication, individually or in different combinations. Based on the in vitro data, our third aim is to apply these two novel approaches directed towards the evaluation of antibody blocking and immunoreconstitution in a SCID mouse animal model of HIV-1.