Inhibition of Human Immunodeficiency Virus-1 (HIV-1) Replication After Transduction of Granulocyte Colony-Stimulating Factor–Mobilized CD34+ Cells From HIV-1–Infected Donors Using Retroviral Vectors Containing Anti–HIV-1 Genes

Abstract

Abstract Transfer of “anti-HIV-1 genes” into hematopoietic stem cells of human immunodeficiency virus-1 (HIV-1)–infected individuals may be a potent therapeutic approach to render mature cells arising from transduced stem cells resistant to the destructive events associated with HIV-1 infection. To determine the feasibility of gene therapy for acquired immunodeficiency syndrome in individuals already infected with HIV-1, granulocyte colony-stimulating factor mobilized peripheral blood CD34+ cells were isolated from HIV-1–infected individuals and transduced with retroviral vectors containing three different anti–HIV-1-genes: the Rev binding domain of the Rev Responsive Element (RRE decoy) (L-RRE-neo), a double hammerhead ribozyme vector targeted to cleave the tat and rev transcripts (L-TR/TAT-neo), and the trans-dominant mutant of rev (M10) (L-M10-SN). As a control, a vector mediating only neomycin resistance (LN) was used. After 3 days of transduction on allogeneic stroma in the presence of stem cell factor, interleukin-6 (IL-6), and IL-3, the cultures were G418-selected, and then challenged with HIV-1JR-FL and a primary HIV-1 isolate. Compared with the control cultures, the L-RRE-neo–, L-TR/TAT-neo–, and L-M10-SN–transduced cultures displayed up to 1,000-fold inhibition of HIV-1 replication after challenge with HIV-1JR-FL and the primary HIV-1 isolate. Growth of the hematopoietic cells in long-term bone marrow culture was not perturbed by the presence of any of the anti–HIV-1 genes. This study shows that anti–HIV-1 genes can be introduced into CD34+ cells from individuals already infected with HIV-1, and strongly inhibit HIV-1 replication in primary monocytes derived from the CD34+ progenitors.