title: Simian virus 40-based replication of catalytically inactive human immunodeficiency virus type 1 integrase mutants in nonpermissive T cells and monocyte-derived macrophages.
abstract: 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.
authors.affiliation: Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Lu
authors.forename: Noriko
authors.initials: N
authors.lastname: Nakajima
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hofmann
authors.forename: Monsef
authors.initials: M
authors.lastname: Benkirane
authors.forename: Kuan-Teh
authors.initials: KT
authors.lastname: Jeang
authors.forename: Joseph
authors.initials: J
authors.lastname: Sodroski
authors.forename: Alan
authors.initials: A
authors.lastname: Engelman
authors.forename: Kuan
authors.initials: K
authors.lastname: Teh-Jeang
mesh_terms: 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
chemical_list: D000952:Antigens, Polyomavirus Transforming; D011993:Recombinant Fusion Proteins; D019427:HIV Integrase