A novel real‐time CTL assay to measure designer T‐cell function against HIV Env+ cells. Issue 5 (20th August 2014)

• Record Type: Journal Article
• Authors: MacLean, Andrew G.; Walker, Edith; Sahu, Gautam K.; Skowron, Gail; Marx, Preston; von Laer, Dorothee; Junghans, Richard P.; Braun, Stephen E.
• Is Part Of: Journal of medical primatology. Volume 43:Issue 5(2014)
• Page Start: 341
• Page End: 348
• Publication Date: 2014-08-20
• Subjects: Primates -- Periodicals; Primates -- Diseases -- Periodicals; Medicine, Experimental -- Periodicals; 616.02738
• DOI: 10.1111/jmp.12137
• Languages: English
• ISSNs: 0047-2565

Abstract

<abstract abstract-type="main" id="jmp12137-abs-0001"> <title>Abstract</title> <sec id="jmp12137-sec-0001" sec-type="section"> <title>Background</title> <p>To increase the immunosurveillance in HIV infection, we used retroviral vectors expressing CD4‐chimeric antigen receptors (CARs) to genetically modify autologous T cells and redirect CTL toward HIV. The CD4 extracellular domain targets envelope and the intracellular signaling domains activate T cells. The maC46 fusion inhibitor binds HIV and blocks viral replication.</p> </sec> <sec id="jmp12137-sec-0002" sec-type="section"> <title>Methods</title> <p>We stimulated rhesus PBMCs with antibodies to CD3/CD28 and cotransduced T cells with CD4‐CAR and maC46 vectors. CD4‐CAR‐transduced T cells were added to Env⁺ 293T cells at E:T of 1:1. Killing of target cells was measured as reduced impedance.</p> </sec> <sec id="jmp12137-sec-0003" sec-type="section"> <title>Results</title> <p>We observed gene expression in 60–70% of rhesus CD3⁺CD8⁺ T cells with the individual vectors and in 35% of the cells with both vectors. CD4‐CAR‐transduced populations specifically killed Env⁺ cells.</p> </sec> <sec id="jmp12137-sec-0004" sec-type="section"> <title>Conclusions</title> <p>In these studies, we showed that designer T cells were redirected to kill Env⁺ cells. Control of viremia without HAART would revolutionize treatment for HIV patients.</p> </sec> </abstract>