Liu D, Donegan J, Kelker N, Nuovo G, Laurence J; International Conference on AIDS.
Int Conf AIDS. 1996 Jul 7-12; 11: 24 (abstract no. LB.A.6012).
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.
Publication Types:
Keywords:
- Acquired Immunodeficiency Syndrome
- Antigens, CD4
- Base Sequence
- CD4-Positive T-Lymphocytes
- Gene Products, tat
- Genes, tat
- HIV Infections
- HIV Seropositivity
- HIV-1
- RNA, Antisense
- RNA, Small Nuclear
- genetics
- immunology
Other ID:
UI: 102225462
From Meeting Abstracts