Selective killing of human immunodeficiency virus-infected cells by targeted gene transfer and inducible gene expression using a recombinant human immunodeficiency virus vector.
A human immunodeficiency virus type 1 (HIV-1)-based retroviral vector pseudotyped with HIV envelope containing the herpes simplex virus-thymidine kinase (HSV-TK) gene under the control of the HIV LTR promoter (pHXTKN) was constructed and stably transferred into human CD4(+) H9, CEM, and U937 cells. RNase protection assays did not initially detect expression of the HSV-TK gene in HXTKN-transduced CD4(+) cells (HXTKN/CD4), but expression was then efficiently induced by infection with HIV-1. MTT assays showed that after HIV-1 infection, the susceptibility of HXTKN/CD4 cells to ganciclovir (GCV) was 1000-fold higher than prior to infection. This enabled HIV-1-infected cells to be selectively killed by transduction with HXTKN followed by exposure to GCV. Because the HSV-TK gene is specifically transferred into HIV-1-permissive cells and expressed only after HIV-1 infection, the frequency of unwanted cell death should be low. Elimination of the HIV-1-infected cells effectively inhibited further spread of infectious virus. In addition, the integrated HIV vector sequences were repackaged on infection with HIV-1 and transferred to surrounding untransduced cells. These results are indicative of the potential benefits of using HIV vectors in gene therapies for the treatment of HIV-1 infection.
Miyake
K
K
Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research, Center for Advanced Medical Technology, Nippon Medical School, Tokyo 113-8602, Japan. kmiyake@nms.ac.jp
Iijima
O
O
Suzuki
N
N
Matsukura
M
M
Shimada
T
T
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D019556:COS Cells; D016923:Cell Death; D002460:Cell Line; D004396:Coloring Agents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015774:Ganciclovir / Q000494:pharmacology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D015658:HIV Infections / Q000628:therapy*; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D018139:Simplexvirus / Q000235:genetics; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D013937:Thymidine Kinase / Q000235:genetics; D013997:Time Factors
D000998:Antiviral Agents; D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D013937:Thymidine Kinase; D012260:Ribonucleases; C022616:thiazolyl blue; D015774:Ganciclovir