Abstract
Background
Previously we reported that human CD4^+^ T cell lines stably expressing anti‐HIV‐1 gag p17 scFv/Cκ in the cytosol or nucleus were resistant to HIV‐1 challenge. Inhibition of HIV‐1 by anti‐HIV‐1 gag p17 scFv/Cκ occurred at both the pre‐ and post‐integration steps of the viral cycle. To simulate more closely the in vivo infection process, in this study we tested anti‐HIV‐1 activity of anti‐HIV‐1 gag p17 scFv/Cκ in primary human T cells.
Methods
Anti‐HIV‐1 gag p17 scFv/Cκ gene that is targeted into cytoplasm was inserted into a MMLV vector and transfected into packaging cell line PT67. The recombinant virus was used to transduce primary human T cells and human CD4^+^ T cell line Jurkat. Following transduction, transduction efficiency, transgene expression, and cell phenotypes were studied. Transduced cells were then challenged with 100 TCID~50~ of HIV‐1 IIIB and primary isolate 5AO12. Following challenge, HIV‐1 replication was monitored by p24 production.
Results
Both transduced Jurkat and primary human T cells expressed the transgene. The expression of the transgene did not alter cell growth and CD4 or CD8 expression. However, HIV‐1 replication in scFv/Cκ‐transduced Jurkat cells was inhibited by nearly 90% as compared with vector controls. More importantly, HIV‐1 replication in primary human T cells from multiple donors transduced with the anti‐HIV‐1 gag p17 scFv/Cκ gene was inhibited by as much as 99% as compared with primary T cells transduced with the vector control. The inhibition of replication was not due to interference in viral entry or reverse transcription. The less that HIV‐1 replicated in different donor cells, the higher the degree of protection.
Conclusions
The expression of the anti‐HIV‐1 gag p17 scFv/Cκ gene construct in primary human T cells renders these cells resistant to HIV‐1 and points to the potential clinical usefulness of this gene construct for anti‐HIV‐1 gene therapy. Copyright © 2002 John Wiley & Sons, Ltd.