Two Retroviruses Packaged in One Cell Line can Combined Inhibit the Replication of HIV-1 in TZM-bl Cells <sup>*</sup>

Zhipin Liang; Zhiyuan Guo; Xin Wang; Xiaohong Kong; Chang Liu

doi:10.1007/s12250-012-3263-8

December 2012

Zhipin Liang, Zhiyuan Guo, Xin Wang, Xiaohong Kong and Chang Liu. Two Retroviruses Packaged in One Cell Line can Combined Inhibit the Replication of HIV-1 in TZM-bl Cells *[J]. Virologica Sinica, 2012, 27(6): 339-344. doi: 10.1007/s12250-012-3263-8

Citation: Zhipin Liang, Zhiyuan Guo, Xin Wang, Xiaohong Kong, Chang Liu. Two Retroviruses Packaged in One Cell Line can Combined Inhibit the Replication of HIV-1 in TZM-bl Cells ^* .VIROLOGICA SINICA, 2012, 27(6) : 339-344. http://dx.doi.org/10.1007/s12250-012-3263-8

Two Retroviruses Packaged in One Cell Line can Combined Inhibit the Replication of HIV-1 in TZM-bl Cells

cstr: 32224.14.s12250-012-3263-8

1.
School of Medicine, Nankai University, Tianjin 300071, China; 2Tianjin Centers for Disease Control and Prevention, Tianjin 300171, China

通讯作者： Xiaohong Kong, kongxh@nankai.edu.cn; Chang Liu, changliu@nankai.edu.cn
收稿日期： 2012-06-13
录用日期： 2012-11-07

摘要

The cellular protein tetherin tethers the HIV-1 viral particles on the cellular membrane to inhibit the replication of HIV-1. However, the HIV-1 accessory protein Vpu counteracts the antiviral function of tetherin. In this study, two retroviral vector plasmids were constructed. One inhibited the vpu gene expression; the other one over-expressed the tetherin. Both retroviral vector plasmids could be packaged in the packaging cell line PT67 to obtain the corresponding retroviruses. The retroviral vector plasmids' functions of tetherin over-expression or vpu-RNAi were detected at the cell level. Retroviral vector plasmids were transfected to PT67 cells at different ratios from 0T3V to 3T0V, and then mixed retroviruses were harvested. The antiviral functions of mixed retroviruses were detected in HIV-1 infected TZM-bl cells. The results showed that packaged mixed retroviruses could repress the replication of HIV-1 in TZM-bl cells.
- HIV-1
- , Vpu
- , Tetherin
- , Gene therapy

Two Retroviruses Packaged in One Cell Line can Combined Inhibit the Replication of HIV-1 in TZM-bl Cells ^*

1.
School of Medicine, Nankai University, Tianjin 300071, China
2.
Tianjin Centers for Disease Control and Prevention, Tianjin 300171, China

Corresponding author: Xiaohong Kong, kongxh@nankai.edu.cn Chang Liu, changliu@nankai.edu.cn
Received Date: 13 June 2012
Accepted Date: 07 November 2012

Fund Project: National Training Programs of Innovation for Undergraduates 111005505The Fundamental Research Funds for the Central Universities 65011871National Natural Science Foundation of China 30970162National Natural Science Foundation of China 81101245

Abstract

The cellular protein tetherin tethers the HIV-1 viral particles on the cellular membrane to inhibit the replication of HIV-1. However, the HIV-1 accessory protein Vpu counteracts the antiviral function of tetherin. In this study, two retroviral vector plasmids were constructed. One inhibited the vpu gene expression; the other one over-expressed the tetherin. Both retroviral vector plasmids could be packaged in the packaging cell line PT67 to obtain the corresponding retroviruses. The retroviral vector plasmids' functions of tetherin over-expression or vpu-RNAi were detected at the cell level. Retroviral vector plasmids were transfected to PT67 cells at different ratios from 0T3V to 3T0V, and then mixed retroviruses were harvested. The antiviral functions of mixed retroviruses were detected in HIV-1 infected TZM-bl cells. The results showed that packaged mixed retroviruses could repress the replication of HIV-1 in TZM-bl cells.
- HIV-1
- , Vpu
- , Tetherin
- , Gene therapy

References
1. Andrew A J, Miyagi E, Strebel K. 2010. Differential effects of human immunodeficiency virus type 1 vpu on the stability of bst-2/tetherin. J Virol, 85(6): 2611-2619.
2. Douglas J L, Viswanathan K, McCarroll M N, et al. 2009. Vpu directs the degradation of the human immunodeficiency virus restriction factor bst-2/tetherin via a {beta}trcp-dependent mechanism. J Virol, 83(16): 7931-7947.
  doi: 10.1128/JVI.00242-09
3. Goto T, Kennel S J, Abe M, et al. 1994. A novel membrane antigen selectively expressed on terminally differentiated human b cells. Blood, 84(6): 1922-1930.
4. Gupta R K, Mlcochova P, Pelchen-Matthews A, et al. 2009. Simian immunodeficiency virus envelope glycoprotein counteracts tetherin/bst-2/cd317 by intracellular sequestration. Proc Natl Acad Sci U S A, 106(49): 20889-20894.
  doi: 10.1073/pnas.0907075106
5. Hauser H, Lopez L A, Yang S J, et al. 2010. Hiv-1 vpu and hiv-2 env counteract bst-2/tetherin by sequestration in a perinuclear compartment. Retrovirology, 7: 51.
  doi: 10.1186/1742-4690-7-51
6. Ishikawa J, Kaisho T, Tomizawa H, et al. 1995. Molecular cloning and chromosomal mapping of a bone marrow stromal cell surface gene, bst2, that may be involved in pre-b-cell growth. Genomics, 26(3): 527-534.
  doi: 10.1016/0888-7543(95)80171-H
7. Klimkait T, Strebel K, Hoggan M D, et al. 1990. The human immunodeficiency virus type 1-specific protein vpu is required for efficient virus maturation and release. J Virol, 64(2): 621-629.
8. Kong X, West J T, Zhang H, et al. 2008. The human immunodeficiency virus type 1 envelope confers higher rates of replicative fitness to perinatally transmitted viruses than to nontransmitted viruses. J Virol, 82(23): 11609-11618.
  doi: 10.1128/JVI.00952-08
9. Kupzig S, Korolchuk V, Rollason R, et al. 2003. Bst-2/hm1.24 is a raft-associated apical membrane protein with an unusual topology. Traffic, 4(10): 694-709.
  doi: 10.1034/j.1600-0854.2003.00129.x
10. Le Tortorec A, Willey S, Neil S J. 2011. Antiviral inhibition of enveloped virus release by tetherin/bst-2: Action and counteraction. Viruses, 3(5): 520-540.
11. Liu C, Shi Y J, Xuan C h, et al. 2008. Establishment of an in vitro protein modification system with antiserum against ubiquitin-like modifier bisg15. Chin J Biochem Mol Biol, 24(1): 30-34. (in Chinese)
12. Liu C, Chang R, Yao X, et al. 2009. Isg15 expression in response to double-stranded rna or lps in cultured fetal bovine lung (fbl) cells. Vet Res Commun, 33(7): 723-733.
  doi: 10.1007/s11259-009-9221-8
13. Malim M H, Emerman M. 2008. HIV-1 accessory proteins--ensuring viral survival in a hostile environment. Cell Host Microb, 3(6): 388-398.
  doi: 10.1016/j.chom.2008.04.008
14. Miller A D, Chen F. 1996. Retrovirus packaging cells based on 10a1 murine leukemia virus for production of vectors that use multiple receptors for cell entry. J Virol, 70(8): 5564-5571.
15. Naito Y, Ui-Tei K, Nishikawa T, et al. 2006. Sivirus: Web-based antiviral sirna design software for highly divergent viral sequences. Nucleic Acids Res, 34(Web Server issue): W448-450.
16. Neil S J, Zang T, Bieniasz P D. 2008. Tetherin inhibits retrovirus release and is antagonized by hiv-1 vpu. Nature, 451(7177): 425-430.
  doi: 10.1038/nature06553
17. Nguyen D H, Hildreth J E. 2000. Evidence for budding of human immunodeficiency virus type 1 selectively from glycolipid-enriched membrane lipid rafts. J Virol, 74(7): 3264-3272.
  doi: 10.1128/JVI.74.7.3264-3272.2000
18. Platt E J, Bilska M, Kozak S L, et al. 2009. Evidence that ecotropic murine leukemia virus contamination in tzm-bl cells does not affect the outcome of neutralizing antibody assays with human immunodeficiency virus type 1. J Virol, 83(16): 8289-8292.
  doi: 10.1128/JVI.00709-09
19. Reinke S O, Bayer M, Berger M, et al. 2011. Analysis of cell surface n-glycosylation of the human embryonic kidney 293t cell line. J Carbohydr Chem, 30(4-6): 218-232.
  doi: 10.1080/07328303.2011.600489
20. Salmon P, Trono D. 2006. Design and production of human immunodeficiency virus-derived vectors, In: Cell biology: A laboratory handbook. Celis J E, ed. , Third Edition: Elsevier Inc. Volume 1, pp425-434.
21. Skasko M, Wang Y, Tian Y, et al. 2011. HIV-1 vpu protein antagonizes innate restriction factor bst-2 via lipid-embedded helix-helix interactions. J Biol Chem, 287(1): 58-67.
22. Terwilliger E F, Cohen E A, Lu Y C, et al. 1989. Functional role of human immunodeficiency virus type 1 vpu. Proc Natl Acad Sci U S A, 86(13): 5163-5167.
  doi: 10.1073/pnas.86.13.5163
23. Tokarev A, Skasko M, Fitzpatrick K, et al. 2009. Antiviral activity of the interferon-induced cellular protein bst-2/tetherin. AIDS Res Hum Retroviruses, 25(12): 1197-1210.
  doi: 10.1089/aid.2009.0253
24. Van Damme N, Goff D, Katsura C, et al. 2008. The interferon-induced protein bst-2 restricts hiv-1 release and is downregulated from the cell surface by the viral vpu protein. Cell Host Microbe, 3(4): 245-252.
  doi: 10.1016/j.chom.2008.03.001
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