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Identification and characterization of differentially expressed mRNAs in HIV type 1-infected human T cells
1Department of Microbiology, School of Medicine, Tokyo Medical and Dental University, Japan.
Abstract:
We used a novel differential display (DD) technique to identify host factors involved in virus replication, pathogenesis, and host response in HIV-1-infected T cells. Thirteen cDNA fragments differentially expressed in HIV-1NL4-3-infected MT-4 cells prior to the occurrence of specific apoptotic cell death were sequenced and identified. Two of seven elevated genes were identical to HIV-1 sequences and the other five were MIP-1alpha, ACTE-III, CD11c, arginase I, and CCR5. The six downregulated genes included prothymosin-a, Jaw-1, proteasome subunit XAPC7, splicing factor 9G8, GA17 protein, and an unknown mRNA. Northern blot and RT-PCR analyses confirmed the altered gene expressions in MT-4 cells as well as in another T cell line, MOLT-4. We also revealed that the amount of MIP-1alpha in culture supernatant of HIV-1-infected cells was increased by more than 15-fold relative to control cells, and the expression of its receptor CCR5 was cooperatively upregulated on the surface of these cells. Furthermore, the upregulation of CD11c after HIV-1 infection was slightly inhibited by blocking the MIP-1alpha-mediated signal transduction. These results indicate that genes altered on HIV-1 infection may be mutually organized and play an important role in HIV-1-induced pathogenesis.
Insights
Researchers identified key host genes altered during HIV-1 infection using differential display. Upregulated genes like MIP-1alpha and CCR5, and downregulated genes, suggest coordinated roles in viral pathogenesis.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Human Immunodeficiency Virus type 1 (HIV-1) infection impacts T cells, influencing replication, pathogenesis, and host response.
- Understanding host factors involved in HIV-1 pathogenesis is crucial for developing effective therapies.
Purpose of the Study:
- To identify host genes differentially expressed in HIV-1-infected T cells using a novel differential display (DD) technique.
- To elucidate the roles of these altered genes in HIV-1 replication, pathogenesis, and host response.
Main Methods:
- Employed a novel differential display (DD) technique to analyze gene expression in HIV-1-infected MT-4 cells.
- Sequenced and identified thirteen differentially expressed cDNA fragments.
- Confirmed gene expression changes using Northern blot and RT-PCR analyses in MT-4 and MOLT-4 cell lines.
- Investigated the functional relationship between MIP-1alpha, CCR5, and CD11c expression post-HIV-1 infection.
Main Results:
- Identified five upregulated genes (MIP-1alpha, ACTE-III, CD11c, arginase I, CCR5) and six downregulated genes (prothymosin-a, Jaw-1, proteasome subunit XAPC7, splicing factor 9G8, GA17 protein, unknown mRNA).
- Observed a >15-fold increase in MIP-1alpha in the supernatant of HIV-1-infected cells, with cooperative upregulation of its receptor CCR5 on cell surfaces.
- Demonstrated that MIP-1alpha signaling partially mediates CD11c upregulation following HIV-1 infection.
Conclusions:
- HIV-1 infection induces significant alterations in host gene expression, including upregulation of inflammatory mediators like MIP-1alpha and its receptor CCR5.
- These differentially expressed genes appear to be interconnected and play a significant role in the pathogenesis of HIV-1.
- The findings provide insights into the molecular mechanisms underlying HIV-1 pathogenesis and potential therapeutic targets.