T-cell protection and enrichment through lentiviral CCR5 intrabody gene delivery

• Published in: Gene therapy Vol. 13; no. 20; pp. 1480 - 1492
• Main Authors: SWAN, C. H, BÜHLER, B, TSCHAN, M. P, BARBAS, C. F, TORBETT, B. E
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 01.10.2006
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Antibodies - genetics; Antibodies - metabolism; Antiviral activity; Applied cell therapy and gene therapy; Biological and medical sciences; Biotechnology; Care and treatment; CCR5 protein; CD34 antigen; CD4 antigen; CD4-Positive T-Lymphocytes - immunology; CD4-Positive T-Lymphocytes - metabolism; CD4-Positive T-Lymphocytes - virology; CD8 antigen; Cell interactions; Cell Line; Cell lines; Cell surface; Cells, Cultured; Chemokines; Cytoprotection; Dendritic cells; Dendritic Cells - immunology; Dendritic Cells - virology; Diabetes mellitus; Effector cells; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression Regulation; Gene targeting; Gene therapy; Gene transfer; Genetic aspects; Genetic Therapy - methods; Health. Pharmaceutical industry; HIV; HIV infection; HIV Infections - immunology; HIV Infections - therapy; HIV-1 - physiology; Human immunodeficiency virus; Human immunodeficiency virus 1; Humans; Immunology; Industrial applications and implications. Economical aspects; Infections; Intrabodies; Lymphocytes; Lymphocytes T; Medical sciences; Methods; Mice; Mice, SCID; Physiological aspects; Receptors, CCR5 - genetics; Receptors, CCR5 - metabolism; Rodents; T cells; Thymocytes; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; United States; Virus; HIV-1; gene delivery; HIV-1 vector; enrichment; CCR5 intrabody; T-Lymphocyte; Retroviridae; Gene therapy; Lentivirus; Protection
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/sj.gt.3302801

CCR5 is the chemokine co-receptor for R5-tropic human immunodeficiency virus type 1 (HIV-1) isolates most often associated with primary infection. We have developed an HIV-1 self-inactivating vector, CAD-R5, containing a CCR5 single-chain antibody (intrabody) gene, which when expressed in T-cell lines and primary CD4+ T cells disrupts CCR5 cell surface expression and provides protection from R5-tropic isolate exposure. Furthermore, CAD-R5 intrabody expression in primary CD4+ T cells supports significant growth and enrichment over time during HIV-1-pulsed dendritic cell-T-cell interactions. These results indicate that CCR5 intrabody-expressing CD4+ T cells are refractory against this highly efficient primary route of infection. CD34+ cells transduced with the CAD-R5 vector gave rise to CD4+ and CD8+ thymocytes in non-obese diabetic (NOD)/ severely combined-immunodeficient (SCID)-human thymus/liver (hu thy/liv) mice, suggesting that CCR5 intrabody expression can be maintained throughout differentiation without obvious cellular effects. CD4+ T cells isolated from NOD/SCID-hu thy/liv mice were resistant to R5-tropic HIV-1 challenge demonstrating the maintenance of protection. Our findings demonstrate delivery of anti-HIV-1 activity through CCR5 intrabodies in primary CD4+ T cells and CD34+ cell-derived T-cell progeny. Thus, gene delivery strategies that provide a selective survival and growth advantage for T effector cells may provide a therapeutic benefit for HIV-1-infected individuals who have failed conventional therapies.