title: Development of single-cycle replicable human immunodeficiency virus 1 mutants.
abstract: Non-infectious but antigenic human immunodeficiency virus 1 (HIV-1) particles are essential tool for the research on many topics associated with this virus. Here we report the construction of plasmid containing the HIV-1 genome mutated in the pol gene, which was co-transfected with plasmids expressing the pol gene products reverse transcriptase (RT) and integrase (IN), and the glycoprotein G of vesicular stomatitis virus (VSV-G). The virions produced in HEK 293 T cells were antigenic, but able to replicate only for one cycle, e.g. first generation single-cycle replicable (SCR) virions. The presence of VSV-G in the envelope of these virions had to ensure a wider spectrum of susceptible cell types for the replication of SCR. Replication of the first generation SCR virions in HEK 293T, MT-2, and mouse spleen cells was examined by p24-capture ELISA, syncytium formation assay, and electron microscopy (EM). HEK 293T and MT-2 cell lines showed a similar replication capacity, while primary cultures of mouse spleen cells were much less effective. The infection of MT-2 cells with the first generation of SCR virions yielded the second generation SCR virions, which were non-infectious. Summing up, the HIV-1 SCR virions represent the useful tool for HIV-1 research facilitating a better biological safety. Moreover, considering their antigenic composition and limited replication, SCR virions may be a promising candidate for the vaccine studies.
authors: Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehern, Iran.
authors: R
authors: R
authors: Zabihollahi
authors: S M
authors: SM
authors: Sadat
authors: R
authors: R
authors: Vahabpour
authors: M R
authors: MR
authors: Aghasadeghi
authors: A
authors: A
authors: Memarnejadian
authors: T
authors: T
authors: Ghazanfari
authors: M
authors: M
authors: Salehi
authors: A
authors: A
authors: Rezaei
authors: K
authors: K
authors: Azadmanesh
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D016368:Frameshift Mutation; D017353:Gene Deletion; D015751:Genes, pol; D057809:HEK293 Cells; D019427:HIV Integrase / Q000096:biosynthesis / Q000235:genetics; D054303:HIV Reverse Transcriptase / Q000096:biosynthesis / Q000235:genetics; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D014779:Virus Replication / Q000235:genetics*; D054302:pol Gene Products, Human Immunodeficiency Virus / Q000097:blood / Q000235:genetics
chemical_list: C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D054302:pol Gene Products, Human Immunodeficiency Virus; D019427:HIV Integrase; D054303:HIV Reverse Transcriptase