Tang Xiaoli, Lu Huafei, Tarwater Patrick M, Silverberg David L, Schorl Christoph, Ramratnam Bharat
Division of Infectious Diseases, Department of Medicine, Warren Alpert Medical School, Brown University, Providence, RI 02903, USA.
Department of Epidemiology and Biostatistics, Texas A&M School of Public Health, College Station, TX 77843, USA.
Microorganisms. 2024 Aug 18;12(8):1707. doi: 10.3390/microorganisms12081707.
Combinatorial antiretroviral therapy (cART) has transformed HIV infection from a death sentence to a controllable chronic disease, but cannot eliminate the virus. Latent HIV-1 reservoirs are the major obstacles to cure HIV-1 infection. Previously, we engineered exosomal Tat (Exo-Tat) to reactivate latent HIV-1 from the reservoir of resting CD4+ T cells. Here, we present an HIV-1 eradication platform, which uses our previously described Exo-Tat to activate latent virus from resting CD4+ T cells guided by the specific binding domain of CD4 in interleukin 16 (IL16), attached to the N-terminus of exosome surface protein lysosome-associated membrane protein 2 variant B (Lamp2B). Cells with HIV-1 surface protein gp120 expressed on the cell membranes are then targeted for immune cytolysis by a BiTE molecule CD4-αCD3, which colocalizes the gp120 surface protein of HIV-1 and the CD3 of cytotoxic T lymphocytes. Using primary blood cells obtained from antiretroviral treated individuals, we find that this combined approach led to a significant reduction in replication-competent HIV-1 in infected CD4+ T cells in a clonal in vitro cell system. Furthermore, adeno-associated virus serotype DJ (AAV-DJ) was used to deliver Exo-Tat, IL16lamp2b and CD4-αCD3 genes by constructing them in one AAV-DJ vector (the plasmid was named pEliminator). The coculture of T cells from HIV-1 patients with Huh-7 cells infected with AAV-Eliminator viruses led to the clearance of HIV-1 reservoir cells in the in vitro experiment, which could have implications for reducing the viral reservoir in vivo, indicating that Eliminator AAV viruses have the potential to be developed into therapeutic biologics to cure HIV-1 infection.
联合抗逆转录病毒疗法（cART）已将HIV感染从死刑判决转变为可控的慢性疾病，但无法消除病毒。潜伏的HIV-1储存库是治愈HIV-1感染的主要障碍。此前，我们设计了外泌体Tat（Exo-Tat），以从静息CD4+T细胞储存库中重新激活潜伏的HIV-1。在此，我们提出了一个HIV-1根除平台，该平台利用我们之前描述的Exo-Tat，在白细胞介素16（IL16）中CD4的特异性结合域的引导下，从静息CD4+T细胞中激活潜伏病毒，该结合域附着在外泌体表面蛋白溶酶体相关膜蛋白2变体B（Lamp2B）的N端。然后，通过双特异性T细胞衔接分子CD4-αCD3将细胞膜上表达HIV-1表面蛋白gp120的细胞作为免疫细胞溶解的靶点，该分子使HIV-1的gp120表面蛋白与细胞毒性T淋巴细胞的CD3共定位。使用从接受抗逆转录病毒治疗的个体获得的原代血细胞，我们发现在克隆体外细胞系统中，这种联合方法导致感染的CD4+T细胞中具有复制能力的HIV-1显著减少。此外，通过在一个腺相关病毒血清型DJ（AAV-DJ）载体中构建Exo-Tat、IL16lamp2b和CD4-αCD3基因，利用AAV-DJ来递送这些基因（该质粒命名为pEliminator）。HIV-1患者的T细胞与感染AAV-Eliminator病毒的Huh-7细胞共培养，在体外实验中导致HIV-1储存库细胞的清除，这可能对减少体内病毒储存库有影响，表明Eliminator AAV病毒有潜力被开发成治疗性生物制品以治愈HIV-1感染。
