Li Haishan, Lahusen Tyler, Xiao Lingzhi, Muvarak Nidal, Blazkova Jana, Chun Tae-Wook, Pauza C David
American Gene Technologies International, Rockville, MD, USA.
Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases (NIAID), Bethesda, MD, USA.
Mol Ther Methods Clin Dev. 2020 May 3;17:1048-1060. doi: 10.1016/j.omtm.2020.04.024. eCollection 2020 Jun 12.
Activation, infection, and eventual depletion of human immunodeficiency virus (HIV)-specific cluster of differentiation 4 (CD4) T cells are the crucial pathogenetic events in acquired immunodeficiency syndrome (AIDS). We developed a cell and gene therapy to reconstitute HIV-specific CD4 T cells and prevent their destruction by HIV. Antigen-specific CD4 T cells will provide helper functions to support antiviral cytotoxic T lymphocyte (CTL) function and the production of virus-specific antibodies. However, expansion of HIV-specific CD4 T cells is poor and previous gene therapies focused on bulk CD4 T cells without enriching for an antigen-specific subset. We developed a method for manufacturing autologous CD4 T cell products highly enriched with Gag-specific T cells. Rare Gag-specific CD4 T cells in peripheral blood mononuclear cells (PBMCs) were increased nearly 1,000-fold by stimulating PBMC with Gag peptides, followed by depleting nontarget cells and transducing with lentivirus vector AGT103 to protect against HIV-mediated depletion and inhibit HIV release from latently infected cells. The average percentage of HIV-specific CD4 cells in the final products was 15.13%, and the average yield was 7 × 10 cells. The protocol for clinical-scale manufacturing of HIV-specific and HIV-resistant CD4 T cells is an important step toward effective immunotherapy for HIV disease.
人类免疫缺陷病毒（HIV）特异性分化簇4（CD4）T细胞的激活、感染以及最终耗竭是获得性免疫缺陷综合征（AIDS）发病过程中的关键致病事件。我们开发了一种细胞和基因疗法，以重建HIV特异性CD4 T细胞并防止其被HIV破坏。抗原特异性CD4 T细胞将提供辅助功能，以支持抗病毒细胞毒性T淋巴细胞（CTL）功能以及病毒特异性抗体的产生。然而，HIV特异性CD4 T细胞的扩增效果不佳，并且先前的基因疗法侧重于整体CD4 T细胞，而未富集抗原特异性亚群。我们开发了一种制造高度富集Gag特异性T细胞的自体CD4 T细胞产品的方法。通过用Gag肽刺激外周血单核细胞（PBMC），然后去除非靶细胞并用慢病毒载体AGT103进行转导，以防止HIV介导的耗竭并抑制HIV从潜伏感染细胞中释放，外周血单核细胞中罕见的Gag特异性CD4 T细胞增加了近1000倍。最终产品中HIV特异性CD4细胞的平均百分比为15.13%，平均产量为7×10个细胞。用于临床规模制造HIV特异性和抗HIV CD4 T细胞的方案是迈向HIV疾病有效免疫治疗的重要一步。
