Functional deletion of the CCR5 receptor by intracellular immunization produces cells that are refractory to CCR5-dependent HIV-1 infection and cell fusion.
Studies of naturally occurring polymorphisms of the CCR5 gene have shown that deletion of the functional receptor or reduced expression of the gene can have beneficial effects in preventing HIV-1 infection or delaying disease. Because these polymorphisms are found in otherwise healthy people, strategies that aim to prevent or limit expression of CCR5 should be beneficial in the treatment of HIV-1 disease. To test this approach we have developed a CCR5-specific single-chain antibody that was expressed intracellularly and retained in the endoplasmic reticulum. This CCR5-intrabody efficiently blocked surface expression of human and rhesus CCR5 and thus prevented cellular interactions with CCR5-dependent HIV-1 and simian immunodeficiency virus envelope glycoprotein. Intrabody-expressing cells were shown to be highly refractory to challenge with R5 HIV-1 viruses or infected cells. These results suggest that gene therapy approaches that deliver this intracellular antibody could be of benefit to infected individuals. Because the antibody reacts with a conserved primate epitope on CCR5 this strategy can be tested in nonhuman lentivirus models of HIV-1 disease.
Steinberger
P
P
Skaggs Institute for Chemical Biology and Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Andris-Widhopf
J
J
Bühler
B
B
Torbett
B E
BE
Barbas
C F
CF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology / Q000502:physiology; D019556:COS Cells; D002459:Cell Fusion / Q000276:immunology; D002460:Cell Line; D005786:Gene Expression Regulation; D015686:Gene Products, env / Q000502:physiology; D015658:HIV Infections / Q000517:prevention & control* / Q000821:virology; D015497:HIV-1 / Q000254:growth & development; D006801:Humans; D007114:Immunization; D008253:Macaca mulatta; D010957:Plasmids / Q000235:genetics; D019713:Receptors, CCR5 / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D011956:Receptors, Cell Surface / Q000235:genetics; D014162:Transfection
D000911:Antibodies, Monoclonal; D015686:Gene Products, env; D019713:Receptors, CCR5; D011956:Receptors, Cell Surface