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Replication of primary HIV-1 isolates is inhibited in PM1 cells expressing sCD4-KDEL
S Degar1, J E Johnson, E Boritz
1Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Abstract:
Expression of a soluble CD4 molecule (sCD4-KDEL) containing a specific retention signal for the endoplasmic reticulum was shown previously to block propagation of the HIV-1MN prototype strain in a transformed T cell line. However, the virus present in HIV-1-infected individuals is more closely represented by primary HIV-1 isolates which, unlike the HIV-1MN strain, have not been adapted to growth in cell lines. To determine if sCD4-KDEL could block replication of primary isolates we used the PM1 cell line that has been shown to propagate primary isolates without adaptation. Here we show that the replication of four primary HIV-1 isolates was strongly inhibited in PM1 cells that expressed sCD4-KDEL under control of the HIV-1 LTR. Infection with primary HIV-1 isolates induced sCD4-KDEL expression driven by the LTR, HIV-1 spread was dramatically reduced, and reverse transcriptase activity in the cell culture supernatants was greatly diminished sCD4-KDEL, therefore, represents a potent inhibitor of HIV-1 replication for gene therapy-based approaches for the treatment of AIDS.
Insights
Soluble CD4 (sCD4-KDEL) effectively inhibits replication of primary HIV-1 isolates in cell lines. This finding supports gene therapy strategies for Acquired Immunodeficiency Syndrome (AIDS) treatment.
Area of Science:
- Immunology
- Virology
- Gene Therapy
Background:
- Previous studies showed soluble CD4 (sCD4-KDEL) blocked HIV-1MN strain propagation.
- Primary HIV-1 isolates, more representative of infections, differ from cell-line-adapted strains.
- The PM1 cell line propagates primary HIV-1 isolates without adaptation.
Purpose of the Study:
- To evaluate sCD4-KDEL's efficacy against primary HIV-1 isolates.
- To assess the potential of sCD4-KDEL for gene therapy in AIDS treatment.
Main Methods:
- Utilized the PM1 cell line for primary HIV-1 isolate propagation.
- Expressed sCD4-KDEL under the control of the HIV-1 Long Terminal Repeat (LTR).
- Monitored HIV-1 replication by measuring reverse transcriptase activity.
Main Results:
- Replication of four primary HIV-1 isolates was strongly inhibited in PM1 cells expressing sCD4-KDEL.
- HIV-1 infection induced sCD4-KDEL expression driven by the LTR.
- HIV-1 spread was dramatically reduced, with diminished reverse transcriptase activity.
Conclusions:
- sCD4-KDEL is a potent inhibitor of primary HIV-1 replication.
- sCD4-KDEL demonstrates promise for gene therapy-based treatment of AIDS.