Rev依赖性指示性T细胞系。
Rev-dependent indicator T cell line.
作者信息
Wu Yuntao, Beddall Margaret H, Marsh Jon W
机构信息
Section on Molecular Virology, Laboratory of Cellular and Molecular Regulation, NIMH, Bethesda, MD, USA.
出版信息
Curr HIV Res. 2007 Jul;5(4):394-402. doi: 10.2174/157016207781024018.
Abstract
Measuring virion infectivity is critical for studying and monitoring the process of HIV-1 infection. The easiest and the most common method utilizes reporter cell lines based on the HIV LTR promoter. The early HIV gene product Tat amplifies expression from the LTR; however, there is a background transcriptional activity that is independent of Tat. Furthermore, LTR activity can be influenced by cellular activation states. We have recently constructed a Rev-dependent expression vector, and as a test of this construct's functionality, we have integrated this vector into a continuous T cell line. This novel indicator cell has no measurable background signal, is not affected by elevated metabolic states, and yet responds robustly to the presence of HIV. The line is able to complete TCID50 assays in 3-5 days, and appears sensitive to both CCR5- and CXCR4-utilizing viruses.
摘要
测量病毒体感染性对于研究和监测HIV-1感染过程至关重要。最简单且最常用的方法是利用基于HIV LTR启动子的报告细胞系。早期HIV基因产物Tat可增强LTR的表达；然而，存在独立于Tat的背景转录活性。此外，LTR活性会受到细胞激活状态的影响。我们最近构建了一种依赖Rev的表达载体，作为对该构建体功能的测试，我们已将此载体整合到一个连续T细胞系中。这种新型指示细胞没有可测量的背景信号，不受代谢状态升高的影响，但对HIV的存在有强烈反应。该细胞系能够在3至5天内完成TCID50测定，并且对利用CCR5和CXCR4的病毒均表现出敏感性。
相似文献
1
Rev-dependent indicator T cell line.
Curr HIV Res. 2007 Jul;5(4):394-402. doi: 10.2174/157016207781024018.
2
Rev-dependent lentiviral expression vector.
Retrovirology. 2007 Feb 7;4:12. doi: 10.1186/1742-4690-4-12.
3
Defective lentiviral vectors are efficiently trafficked by HIV-1 and inhibit its replication.
Mol Ther. 2001 Jun;3(6):928-39. doi: 10.1006/mthe.2001.0344.
4
Inhibition of human immunodeficiency virus type 1 by Tat/Rev-regulated expression of cytosine deaminase, interferon alpha2, or diphtheria toxin compared with inhibition by transdominant Rev.
Hum Gene Ther. 1999 Jan 1;10(1):103-12. doi: 10.1089/10430349950019237.
5
Specific marking of HIV-1 positive cells using a Rev-dependent lentiviral vector expressing the green fluorescent protein.
J Vis Exp. 2010 Sep 23(43):2198. doi: 10.3791/2198.
6
Long-term protection against HIV-1 infection conferred by tat or rev antisense RNA was affected by the design of the retroviral vector.
Virology. 1996 Jun 15;220(2):377-89. doi: 10.1006/viro.1996.0326.
7
Expression kinetics and subcellular localization of HIV-1 regulatory proteins Nef, Tat and Rev in acutely and chronically infected lymphoid cell lines.
Arch Virol. 1994;139(3-4):365-78. doi: 10.1007/BF01310798.
8
A combination anti-HIV-1 gene therapy approach using a single transcription unit that expresses antisense, decoy, and sense RNAs, and trans-dominant negative mutant Gag and Env proteins.
Front Biosci. 2002 Feb 1;7:a15-28. doi: 10.2741/ding.
9
An emerging and variant viral promoter of HIV-1 subtype C exhibits low-level gene expression noise.
Retrovirology. 2021 Sep 19;18(1):27. doi: 10.1186/s12977-021-00572-2.
10
Down-modulation of TCR/CD3 surface complexes after HIV-1 infection is associated with differential expression of the viral regulatory genes.
Eur J Immunol. 2001 Apr;31(4):969-79. doi: 10.1002/1521-4141(200104)31:4<969::aid-immu969>3.0.co;2-2.
引用本文的文献
1
High-throughput High Content Quantification of HIV-1 Viral Infectious Output.
bioRxiv. 2025 Jul 1:2025.06.28.662157. doi: 10.1101/2025.06.28.662157.
2
Phenotypic Characterization of Replication-Impaired Lenacapavir-Resistant HIV Clinical Isolates.
J Med Virol. 2025 Apr;97(4):e70340. doi: 10.1002/jmv.70340.
3
CD34 serves as an intrinsic innate immune guardrail protecting stem cells from replicating retroviruses.
bioRxiv. 2025 Mar 17:2025.03.15.643450. doi: 10.1101/2025.03.15.643450.
4
Characterization of a CXCR4 antagonist TIQ-15 with dual tropic HIV entry inhibition properties.
PLoS Pathog. 2024 Aug 15;20(8):e1012448. doi: 10.1371/journal.ppat.1012448. eCollection 2024 Aug.
5
Suppression of viral rebound by a Rev-dependent lentiviral particle in SIV-infected rhesus macaques.
Gene Ther. 2025 Jan;32(1):16-24. doi: 10.1038/s41434-024-00467-9. Epub 2024 Jul 18.
6
Discovery of Ten Anti-HIV Hit Compounds and Preliminary Pharmacological Mechanisms Studies.
Curr HIV Res. 2024;22(2):82-90. doi: 10.2174/011570162X301289240320082840.
7
HIV Preintegration Transcription and Host Antagonism.
Curr HIV Res. 2023;21(3):160-171. doi: 10.2174/1570162X21666230621122637.
8
Cutting edge strategies for screening of novel anti-HIV drug candidates against HIV infection: A concise overview of cell based assays.
Heliyon. 2023 May 5;9(5):e16027. doi: 10.1016/j.heliyon.2023.e16027. eCollection 2023 May.
9
Antiviral Properties of HIV-1 Capsid Inhibitor GSK878.
Antimicrob Agents Chemother. 2023 May 17;67(5):e0169422. doi: 10.1128/aac.01694-22. Epub 2023 Apr 11.
10
Identification of the SHREK Family of Proteins as Broad-Spectrum Host Antiviral Factors.
Viruses. 2021 May 4;13(5):832. doi: 10.3390/v13050832.
本文引用的文献
1
Rev-dependent lentiviral expression vector.
Retrovirology. 2007 Feb 7;4:12. doi: 10.1186/1742-4690-4-12.
2
Nef does not affect the efficiency of human immunodeficiency virus type 1 fusion with target cells.
J Virol. 2003 Oct;77(19):10645-50. doi: 10.1128/jvi.77.19.10645-10650.2003.
3
An adenovirus-based fluorescent reporter vector to identify and isolate HIV-infected cells.
J Virol Methods. 2002 Jan;99(1-2):9-21. doi: 10.1016/s0166-0934(01)00375-5.
4
HIV Nef-mediated cellular phenotypes are differentially expressed as a function of intracellular Nef concentrations.
J Biol Chem. 2001 Aug 31;276(35):32763-70. doi: 10.1074/jbc.M101025200. Epub 2001 Jul 3.
5
A luciferase-reporter gene-expressing T-cell line facilitates neutralization and drug-sensitivity assays that use either R5 or X4 strains of human immunodeficiency virus type 1.
Virology. 2001 Feb 15;280(2):292-300. doi: 10.1006/viro.2000.0780.
6
Use of calibrated viral load standards for group M subtypes of human immunodeficiency virus type 1 to assess the performance of viral RNA quantitation tests.
J Clin Microbiol. 2000 Mar;38(3):1247-9. doi: 10.1128/JCM.38.3.1247-1249.2000.
7
A cell line-based neutralization assay for primary human immunodeficiency virus type 1 isolates that use either the CCR5 or the CXCR4 coreceptor.
J Virol. 1999 Nov;73(11):8966-74. doi: 10.1128/JVI.73.11.8966-8974.1999.
8
Cell cycle- and Vpr-mediated regulation of human immunodeficiency virus type 1 expression in primary and transformed T-cell lines.
J Virol. 1999 Jul;73(7):5422-30. doi: 10.1128/JVI.73.7.5422-5430.1999.
9
Quantification of CD4, CCR5, and CXCR4 levels on lymphocyte subsets, dendritic cells, and differentially conditioned monocyte-derived macrophages.
Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):5215-20. doi: 10.1073/pnas.96.9.5215.
10
Human lymphoblastoid CD4(+) T cells become permissive to macrophage-tropic strains of human immunodeficiency virus type 1 after passage into severe combined immunodeficient mice through in vivo upregulation of CCR5: in vivo dynamics of CD4(+) T-cell differentiation in pathogenesis of AIDS.
J Virol. 1998 Dec;72(12):10323-7. doi: 10.1128/JVI.72.12.10323-10327.1998.
Zotero 插件