IDENT TOLL LIKE RECEPTORS AGONIST INDUCED PBMC & CD8+TCELLS ANTIHIV MECHANISM

受体激动剂诱导的 PBMC 识别损伤

• 批准号: 8357103
• 负责人: NAWAL BOUKLI
• 金额: $ 7.91万
• 依托单位: UNIVERSIDAD CENTRAL DEL CARIBE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357103
• 关键词: Agonist; Antiviral Agents; Biological Assay; Biomedical Research; CD8B1 gene; Disease; Dose; Drug resistance; Funding; Grant; HIV; HIV-1; Highly Active Antiretroviral Therapy; Immune system; Immunization; Interferon-alpha; Interferons; Liquid substance; National Center for Research Resources; Peripheral Blood Mononuclear Cell; Play; Principal Investigator; Production; Proteins; Proteomics; Recombinant Proteins; Reporting; Research; Research Infrastructure; Resources; Source; T-Lymphocyte; Techniques; Testing; Therapeutic; Time; Toll-like receptors; Two-Dimensional Gel Electrophoresis; United States National Institutes of Health; chemokine; cost; cytokine; immune function; pathogen; protein expression; receptor; research study; resistant strain; response; restoration; two-dimensional

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. It is now accepted that restoration of the immune function using only highly active antiretroviral therapy (HAART) is incomplete. Because of the emergence of drug-resistant strains of HIV-1, therapeutic immunization strategies are needed to reinforce HAART in the treatment of HIV disease. It has been reported that Toll like receptor agonist 7/8 (R-848) induces anti-HIV-1 activity. Toll like receptors (TLR) are evolutionary conserved pathogen receptors that play a key role in innate and adaptive immune system. The TLR have the ability to induce antiviral factors producing cytokines and B-chemokines and triggering IFN (IFN-alpha and Beta) production. We propose to test the hypothesis that TLR agonist 7/8 induced antiviral activity can be differentially determined by a proteomic approach. Specific Aims 1. To determine the differential anti-HIV1 activity in culture fluids from PBMC, CD8+ T cells and CD8+ T cells depleted PBMC treated with TLR agonist R-848 2. To identify and compare differentially expressed proteins in culture fluids with and without antiviral activity using proteomics 3. To validate the antiviral activity of the identified factors stimulated by TLR agonist R-848 using recombinant protein expression technique We are currently performing dose-response assays to determine the optimal TLR agonist concentration. After 9 days of TLR agonist exposure, culture fluids were collected for Quantitative Real Time PCR and proteomics analysis. Preliminary results reveal that 5 ug/mL is the optimal TLR agonist concentration. Two Dimensional Gel Electrophoresis (2D-GE) experiments from culture fluids at 5 ug/mL and 1 ug/mL were performed in triplicates and differentially expressed proteins were evaluated as compared to the control. We believe that the identified proteins will generate a proteomic signature of the TLR agonist-anti HIV-1 response.

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