Abstract

Human immunodeficiency virus type 1 (HIV-1) entry into host cells occurs through interactions between the viral surface envelope glycoprotein (Env) and the host receptor CD4, and co-receptors CCR5 or CXCR4. However, understanding these interactions in membrane contexts remains limited due to low receptor and coreceptor densities in commonly used cell models. To address this, we developed cellular systems with elevated CD4 expression and either CCR5 or CXCR4. Human osteosarcoma cells (HOS), which lack endogenous surface expression of HIV receptor (CD4) and co-receptors (CCR5 or CXCR4), were engineered via lentiviral transduction to overexpress CD4 and either CCR5 or CXCR4. Cells were subjected to four rounds of fluorescence-activated cell sorting to enrich for populations with high surface expression. Receptor and co-receptor expression levels were assessed using immunoblotting, flow cytometry, and indirect immunofluorescence assay. Functional evaluation with HIV-1 pseudovirus showed that CD4/CCR5-expressing cells exhibited increased susceptibility to the R5-tropic virus, linking receptor density to HIV-1 entry efficiency. These engineered HOS cells, upon further optimization, will be useful for examining HIV-1 Env–host interactions and evaluating entry inhibitors or antibodies.

Date of publication

4-2026

Document Type

Thesis

Language

english

Persistent identifier

http://hdl.handle.net/10950/5067

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