Preclinical Development and Clinical-Scale Manufacturing of HIV Gag-Specific, LentivirusModified CD4 T Cells for HIV Functional Cure

Haishan Li,Tyler Lahusen,Lingzhi Xiao,Nidal Muvarak,Jana Blazkova,Tae-Wook Chun,C David Pauza

doi:10.1016/j.omtm.2020.04.024

Abstract

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, ex vivo 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 × 108 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.

Highlights

human immunodeficiency virus (HIV) infection is a chronic disease characterized by ongoing viral replication with gradual exhaustion and destruction of cluster of differentiation 4 (CD4) T lymphocytes
Construction and Evaluation of Lentivirus AGT103 for Blocking HIV Infection and Replication We developed a recombinant lentivirus vector encoding inhibitory RNA targeting the HIV coreceptor chemokine receptor type 5 (CCR5) and HIV sequences within the Vif/Tat coding regions
A CCR5 targeting sequence is embedded within the naturally occurring human miR30, a HIV TAT targeting sequence is embedded within miR185, and a HIV VIF targeting sequence is embedded within miR21

Summary

Introduction

HIV infection is a chronic disease characterized by ongoing viral replication with gradual exhaustion and destruction of cluster of differentiation 4 (CD4) T lymphocytes. The recent developments of autologous T cell therapies for cancer, chronic viral infection, and viral reactivation post-transplant[1,2,3,4,5,6,7] suggest that a similar strategy might be used to control HIV and reduce the dependence on ART.[8] A consistent feature of successful T cell therapies is the enrichment of antigen-specific cells, either through amplification after antigen stimulation or by introducing new antigen receptors to redirect the T cell response. Our strategy is to reconstitute the CD4 T cell population by infusing HIV-specific CD4 T cells that will be durable within the HIV-infected individual due to the introduction of a lentivirus vector that modulates C-C chemokine receptor type 5 (CCR5) levels and decreases the abundance of HIV mRNA and genomic RNA. We devised an efficient process for manufacturing autologous CD4 T cells from HIV-infected individuals that is both specific for HIV and resistant to viral-mediated depletion

Methods

Results

Conclusion