Down-regulation of CXCR4 expression by SDF-KDEL in CD34+ hematopoietic stem cells: An anti-human immunodeficiency virus strategy
| Autor: | Li Sun, Xue-Fan Bai, Ping-Zhong Wang, Jiu-Cong Zhang, Zhansheng Jia, Yongtao Sun, Jian-Qi Lian, Qing-He Nie, Ye Zhang, Jiu-Ping Wang, Xinhong Li, Chang-Xing Huang, Yan Zhuang |
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| Rok vydání: | 2009 |
| Předmět: |
Receptors
CXCR4 medicine.drug_class Genetic enhancement T cell Genetic Vectors HIV Core Protein p24 Down-Regulation Antigens CD34 Biology Monoclonal antibody CXCR4 Viral vector Flow cytometry Transduction Genetic Virology medicine Humans Cells Cultured medicine.diagnostic_test Lentivirus Infant Newborn virus diseases Flow Cytometry Hematopoietic Stem Cells Molecular biology Fusion protein Chemokine CXCL12 Culture Media medicine.anatomical_structure Cell culture HIV-1 |
| Zdroj: | Journal of Virological Methods. 161:30-37 |
| ISSN: | 0166-0934 |
| Popis: | CXCR4 plays an essential role as the first discovered coreceptor for the entry of T cell tropic isolates of HIV-1. Blocking the surface expression of this receptor may be a potential strategy to prevent HIV-1 infection. A lentiviral vector, pLenti6/V5-S-K, expressing a SDF-KDEL fusion protein was constructed and a replication-incompetent lentiviral stock was produced. The lentiviral stock was transduced into CD34 + hHSC and the transient expression of the recombinant protein, SDF-1, was assayed using indirect immunofluorescence. The surface expression of CXCR4 in CD34 + hHSC pretreated with different amounts of recombinant lentiviral vectors was detected by flow cytometric analysis. A marked down-regulation of CXCR4 expression in the cells transduced with recombinant lentiviral vectors pLenti6/V5-S-K was observed by flow cytometry with PE-conjugated anti-human CXCR4 monoclonal antibodies which showed the percentages of the inhibition effects of CXCR4-SDF-1 mediated syncytium formation are presented by concentration. P24 antigen levels of cell culture supernatants were detected on the 4th, 7th, and 10th day, with 10 3 TCID50 HIV-1 infected CD34 + hHSC to evaluate the inhibitory effect of pLenti6/V5-S-K transduction on HIV-1 infection. The cells transfected with pLenti6/V5-S-K had a significant reduction of HIV-1 DP27 infection compared to controls ( P |
| Databáze: | OpenAIRE |
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