Simian virus 40-based replication of catalytically inactive human immunodeficiency virus type 1 integrase mutants in nonpermissive T cells and monocyte-derived macrophages.
Integrase function is required for retroviral replication in most instances. Although certain permissive T-cell lines support human immunodeficiency virus type 1 (HIV-1) replication in the absence of functional integrase, most cell lines and primary human cells are nonpermissive for integrase mutant growth. Since unintegrated retroviral DNA is lost from cells following cell division, we investigated whether incorporating a functional origin of DNA replication into integrase mutant HIV-1 might overcome the block to efficient gene expression and replication in nonpermissive T-cell lines and primary cells. Whereas the Epstein-Barr virus (EBV) origin (oriP) did little to augment expression from an integrase mutant reporter virus in EBV nuclear antigen 1-expressing cells, simian virus 40 (SV40) oriT dramatically enhanced integrase mutant infectivity in T-antigen (Tag)-expressing cells. Incorporating oriT into the nef position of a full-length, integrase-defective virus strain yielded efficient replication in Tag-expressing nonpermissive Jurkat T cells without reversion to an integration-competent genotype. Adding Tag to integrase mutant-oriT viruses yielded 11.3-kb SV40-HIV chimeras that replicated in Jurkat cells and primary monocyte-derived macrophages. Real-time quantitative PCR analyses of Jurkat cell infections revealed that amplified copies of unintegrated DNA likely contributed to SV40-HIV integrase mutant replication. SV40-based HIV-1 integrase mutant replication in otherwise nonpermissive cells suggests alternative approaches to standard integrase-mediated retroviral gene transfer strategies.
Lu
Richard
R
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
Nakajima
Noriko
N
Hofmann
Wolfgang
W
Benkirane
Monsef
M
Jeang
Kuan-Teh
KT
Sodroski
Joseph
J
Engelman
Alan
A
Teh-Jeang
Kuan
K
D000952:Antigens, Polyomavirus Transforming / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D019427:HIV Integrase / Q000235:genetics* / Q000378:metabolism; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D006801:Humans; D019169:Jurkat Cells / Q000821:virology; D008264:Macrophages / Q000821:virology; D009000:Monocytes / Q000821:virology; D009154:Mutation; D011993:Recombinant Fusion Proteins; D018741:Replication Origin; D013539:Simian virus 40 / Q000235:genetics* / Q000502:physiology; D013601:T-Lymphocytes / Q000821:virology; D016662:Virus Integration; D014779:Virus Replication
D000952:Antigens, Polyomavirus Transforming; D011993:Recombinant Fusion Proteins; D019427:HIV Integrase