title: Antibodies that neutralize SIV(mac)251 in T lymphocytes cause interruption of the viral life cycle in macrophages by preventing nuclear import of viral DNA.
abstract: Previous reports from our lab had shown that sera obtained from SIV(mac)-infected animals neutralized SIV(mac) infectivity in CD4(+) T cells but failed to protect monkey primary macrophages from infection with the virus. However, the antibodies could inhibit completion of the viral life cycle in the macrophages at the postentry stage(s). In this report we examined the mechanisms of the late effect of the antibodies. Using monoclonal antibodies (MAbs), we demonstrated that only antibodies to the SIV envelope protein (KK17 and KK42) but not antibody to the viral core protein (FA2) had the same inhibitory effect as that of the anti-SIV sera. To identify the stage of the viral replication cycle that was inhibited by anti-SIV antibodies in macrophages, we used various PCR techniques to study viral entry/reverse transcription (by amplifying the viral gag gene), viral genome nuclear transport (by amplifying 2-LTR circular forms), viral integration (by Alu-PCR assay), and viral protein expression (by RIPA). We found that in macrophage cultures inoculated with SIV(mac)251 that were preincubated with antienvelope MAbs, viral DNA was detected at 8 h postinoculation but the 2-LTR circular forms and integrated viral DNAs were undetectable, and viral proteins were not expressed in these infected macrophages. These results strongly suggested that anti-SIV antibodies inhibited SIV(mac) replication in macrophages by blocking nuclear transport of viral genomes since viral DNA could not be detected in the nuclei of treated cultures. Furthermore, we showed that although viral replication in macrophages was interrupted by the antibodies, when cocultured with permissive T cells, the viral genomes presented in the cytoplasm of the macrophages could readily transfer to T cells during cell-cell contact. Importantly, this transfer could not be prevented by the antibodies. These results might explain the failure of passive antibody immunization against SIV(mac)251--a critical obstacle in AIDS vaccine development.
authors.affiliation: Department of Microbiology, Molecular Genetics, and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66160-7424, USA. wzhuge@kumc.edu
authors.forename: W
authors.initials: W
authors.lastname: Zhuge
authors.forename: F
authors.initials: F
authors.lastname: Jia
authors.forename: G
authors.initials: G
authors.lastname: Mackay
authors.forename: A
authors.initials: A
authors.lastname: Kumar
authors.forename: O
authors.initials: O
authors.lastname: Narayan
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000914:Antibodies, Viral / Q000494:pharmacology*; D016282:Aspartic Acid Endopeptidases / Q000502:physiology; D002450:Cell Communication; D002478:Cells, Cultured; D004279:DNA, Viral / Q000378:metabolism*; D016679:Genome, Viral; D006801:Humans; D008018:Life Cycle Stages / Q000187:drug effects*; D008253:Macaca mulatta; D008264:Macrophages / Q000187:drug effects* / Q000821:virology; D009500:Neutralization Tests; D015302:Simian Immunodeficiency Virus / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D013601:T-Lymphocytes / Q000821:virology; D014759:Viral Envelope Proteins / Q000276:immunology; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D004279:DNA, Viral; D014759:Viral Envelope Proteins; D016282:Aspartic Acid Endopeptidases; C077164:SIV(mac) proteinase