11th International AIDS Conference Vancouver, British Columbia — July 7-12, 1996

Int Conf AIDS 1996 Jul 7-12; 11:24 (abstract no. LB.A.6012)
Liu D, Donegan J, Kelker N, Nuovo G, Laurence J; Enzo Biochem, Inc., Farmingdale, NY. Fax: (212) 856-0878. E-mail: bet3@cornell.edu.

OBJECTIVE: To develop high level and stable resistance to HIV-1 in CD4+ cells as a critical step in the development of ex vivo antisense therapy.

METHODS: We have produced immune cells with stable and high level resistance to HIV-1 by the introduction of an antisense RNA-producing DNA construct. In order to overcome the variability and mutability of the virus, the construct (pNDUIA,B, C) was designed to produce 3 different antisense sequences directed against two functional regions, tar and tat/rev. As an additional strategy to provide a high level of synthesis, stability and a favorable intracellular localization to the antisense transcripts, each HIV-1 antisense sequence was incorporated into the transcript region of UI snRNA.

RESULTS: CD4+ monocytic cells (U937) stably transfected with pNDUIA,B,C transcribed each of the 3 species of UI/HIV antisense. UI/HIV antisense RNA was confined to the nucleus as determined by in situ RT:PCR. The transfected cells demonstrated a high level and stable resistance to HIV-1 as determined by the following criteria. The cells survived 3 successive challenges by HIV-1 (BAL strain). Cells assayed during the third HIV-1 challenge showed no evidence of virus growth as indicated by lack of detectable p24 production, no detectable HIV-1 sequences by PCR assay and an absence of syncytia. The surviving cells retained the CD4+ surface receptor and the resident pNDUIA,B,C DNA construct.

CONCLUSION: Multitargeting UI/HIV antisense confers upon CD4+ cells high level and stable resistance to HIV-1.

960707
LBA6012

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