Enhancement of cytotoxic T lymphocyte activity by dendritic cells loaded with Tat-protein transduction domain-fused hepatitis B virus core antigen

Xiaohua Chen

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Yongsheng Yu,

Yongsheng Yu

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Qingchun Pan,

Qingchun Pan

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Zhenghao Tang,

Zhenghao Tang

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Jinchao Han,

Jinchao Han

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Guoqing Zang,

Corresponding Author

Guoqing Zang

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

*Corresponding author: Tel, 86-21-64369181-8673; Fax, 86-21-64369181; E-mail, [email protected]Search for more papers by this author

Xiaohua Chen,

Xiaohua Chen

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Yongsheng Yu,

Yongsheng Yu

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Qingchun Pan,

Qingchun Pan

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Zhenghao Tang,

Zhenghao Tang

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Jinchao Han,

Jinchao Han

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

Search for more papers by this author

Guoqing Zang,

Corresponding Author

Guoqing Zang

Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai 200233, China

*Corresponding author: Tel, 86-21-64369181-8673; Fax, 86-21-64369181; E-mail, [email protected]Search for more papers by this author

First published: 16 December 2008

https://doi.org/10.1111/j.1745-7270.2008.00487.x

Citations: 1

This work was supported by a grant from the National Natural Science Foundation of China (No. 30571669)

About

PDF

Tools

Share a link

Email
Wechat
Bluesky

Abstract

The protein transduction domain (PTD) of human immunodeficiency virus-1-Tat protein has a unique potency to penetrate the cellular membranes. To synthesize the sequence of Tat-PTD_47–57 and hepatitis B virus core antigen (HBcAg), we spliced thesesequences and linked a fusion gene into the pMAL-c2x vector. The fusion proteins were purified by affinity chromatography and pulsed with bone marrow-derived dendritic cells (DCs), and the transduction of recombinant protein was detected by immunofluorescence antibody assay. Results showed that recombinant PTD-HBcAg could penetrate into DC cytoplasm while recombinant HBcAg was detected on the surface of cells. The percentage of DC surface molecules, such as CD80, CD86 and major histocompatibility complex II, and production of cytokine (IL-12p70) induced by recombinant PTD-HBcAg were significantly higher than those induced by recombinant HBcAg or tumor necrosis factor-α. DCs treated with PTD-HBcAg induced T cells to differentiate into specific cytotoxic T lymphocytes (CTLs) and enhanced the CTL killing response. In conclusion, the expressed and purified PTD-HBcAg fusion protein could penetrate into cells through the plasma membrane, promote DC maturation, and enhance T cells response to generate HBcAg-specific CTLs efficiently.

References

1 Seeger C, Mason WS., Hepatitis B virus biology. Microbiol Mol Biol Rev 2000, 64: 51–68
10.1128/MMBR.64.1.51-68.2000
CAS PubMed Web of Science® Google Scholar
2 Yang SH, Lee CG, Park SH., Correlation of antiviral T-cell responses with suppression of viral rebound in chronic hepatitis B carriers: a proof-of-concept study. Gene Ther 2006, 13: 1110–1117
10.1038/sj.gt.3302751
CAS PubMed Web of Science® Google Scholar
3 Penna A, Chisari FV, Bertoletti A, Missale G, Missale G, Fowler P, Giuberti T et al., Cytotoxic T lymphocytes recognize an HLA-A2-restricted epitope within the hepatitis B virus nucleocapsid antigen. J Exp Med 1991, 174: 1565–1570
10.1084/jem.174.6.1565
CAS PubMed Web of Science® Google Scholar
4 Löhr HF, Krug S, Herr W, Weyer S, Schlaak J, Wölfel T, Gerken G et al., Quantitative and functional analysis of core-specific T-helper cell and CTL activities in acute and chronic hepatitis B. Liver 1998, 18: 405–413
10.1111/j.1600-0676.1998.tb00825.x
CAS PubMed Web of Science® Google Scholar
5 Jonuleit H, Kühn U, Müller G, Steinbrink K, Paragnik L, Schmitt E, Knop J et al., Pro-infammatory cytokines and prostaglandins induce maturation of potent immunostimulatory dendritic cells under fetal calf serum-free conditions. Eur J Immunol 1997, 27: 3135–3142
10.1002/eji.1830271209
CAS PubMed Web of Science® Google Scholar
6 Rehermann B, Pasquinelli C, Mosier SM, Chisari FV., Hepatitis B virus (HBV) sequence variation of cytotoxic T lymphocyte epitopes is not common in patients with chronic HBV infection. J Clin Invest 1995, 96: 1527–1534
10.1172/JCI118191
CAS PubMed Web of Science® Google Scholar
7 Guidotti LG, Ando K, Hobbs MV, Ishikawa T, Runkel L, Schreiber RD, Chisari FV., Cytotoxic T lymphocytes inhibit hepatitis B virus gene expression by a non-cytolytic mechanism in transgenic mice. Proc Natl Acad Sci USA 1994, 91: 3764–3768
10.1073/pnas.91.9.3764
CAS PubMed Web of Science® Google Scholar
8 Zhang Y, Li S, Shan M, Pan X, Zhuang K, He L, Gould K et al., Hepatitis B virus core antigen epitopes presented by HLA-A2 single-chaitrimers induce functional epitope-specific CD8⁺ T-cell responses in HLA-A2.1/Kb transgenic mice. Immunology 2007, 121: 105–112
10.1111/j.1365-2567.2007.02543.x
CAS PubMed Web of Science® Google Scholar
9 Kim DT, Mitchell DJ, Brockstedt DG, Fong L, Nolan GP, Fathman CG, Engleman EG et al., Introduction of soluble proteins into the MHC class I pathway by conjugation to an HIV tat peptide. J Immunol 1997, 159: 1666–1668
CAS PubMed Web of Science® Google Scholar
10 Schirmbeck R, Riedl P, Zurbriggen R, Akira S, Reimann J., Antigenic epitopes fused to cationic peptide bound to oligonucleotides facilitate Toll-like receptor 9-dependent, but CD4⁺ T cell help-independent, priming of CD8⁺ T cells. J Immunol 2003, 171: 5198–5207
10.4049/jimmunol.171.10.5198
CAS PubMed Web of Science® Google Scholar
11 Ho A, Schwarze SR, Mermelstein SJ, Waksman G, Dowdy SF., Synthetic protein transduction domains: enhanced transduction potential in vitro and in vivo. Cancer Res 2001, 61: 474–477
CAS PubMed Web of Science® Google Scholar
12 Caron NJ, Torrente Y, Camirand G, Bujold M, Chapdelaine P, Leriche K, Bresolin N et al., Intracellular delivery of a TateGFP fusion protein into muscle cells. Mol Ther 2001, 3: 310–318
10.1006/mthe.2001.0279
CAS PubMed Web of Science® Google Scholar
13 Bergamo P, Maurano F, D'Arienzo R, David C, Rossi M., Association between activation of phase 2 enzymes and down-regulation of dendritic cell maturation by c9,t11-conjugated linoleic acid. Immunol Lett 2008, 117: 181–190
10.1016/j.imlet.2008.02.001
CAS PubMed Web of Science® Google Scholar
14 Huang Y, Chen Z, Zhou C, Yao H, Li M, Xu C., The modulation of thymosin alpha 1 in the maturation, differentiation and function of murine bone marrow-derived dendritic cells in the absence or presence of tumor necrosis factor-alpha. Int Immunopharmacol 2004, 4: 539–546
10.1016/j.intimp.2004.02.008
CAS PubMed Web of Science® Google Scholar
15 Vives E, Brodin P, Lebleu B., A truncated HIV-1 Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus. J Biol Chem 1997, 272: 16010–16017
10.1074/jbc.272.25.16010
CAS PubMed Web of Science® Google Scholar
16 Dietz GP, Bahr M., Delivery of bioactive molecules into the cell: the Trojan horse approach. Mol Cell Neurosci 2004, 27: 85–131
10.1016/j.mcn.2004.03.005
CAS PubMed Web of Science® Google Scholar
17 Dietz GP, Kilic E, Bahr M., Inhibition of neuronal apoptosis in vitro and in vivo using TAT-mediated protein transduction. Mol Cell Neurosci 2002, 21: 29–37
10.1006/mcne.2002.1165
CAS PubMed Web of Science® Google Scholar
18 Tolia NH, Joshua-Tor L., Strategies for protein coexpression in Escherichia coli. Nat Methods 2006, 3: 55–64
10.1038/nmeth0106-55
CAS PubMed Web of Science® Google Scholar
19 Macatonia SE, Hosken NA, Litton M, Vieira P, Hsieh CS, Culpepper JA, Wysocka M et al., Dendritic cells produce IL-12 and direct the development of Th1 cells from naive CD41 T cells. J Immunol 1995, 154: 5071–5079
CAS PubMed Web of Science® Google Scholar
20 Koch F, Stanzl U, Jennewein P, Janke K, Heufler C, Kämpgen E, Romani N et al., High level IL-12 production by murine dendritic cells up-regulation via MHC class II and CD40 molecules and down-regulation by IL-4 and IL-10. J Exp Med 1996, 184: 741–746
10.1084/jem.184.2.741
CAS PubMed Web of Science® Google Scholar
21 Dobrzanski MJ, Reome JB, Dutton RW., Type 1 and type 2 CD8⁺ effector T cell subpopulations promote long-term tumor immunity and protection to progressively growing tumor. J Immunol 2000, 164: 916–925
10.4049/jimmunol.164.2.916
CAS PubMed Web of Science® Google Scholar
22 Tsuji T, Chamoto K, Funamoto H, Kosaka A, Matsuzaki J, Abe H, Fujio K et al., Critical role of the Th1/Tc1 circuit for the generation of tumor-specific CTL during tumor eradication in vivo by Th1-cell therapy. Cancer Sci 2003, 94: 924–928
10.1111/j.1349-7006.2003.tb01377.x
PubMed Web of Science® Google Scholar

Citing Literature

Volume40, Issue12

December 2008

Pages 996-1004

Enhancement of cytotoxic T lymphocyte activity by dendritic cells loaded with Tat-protein transduction domain-fused hepatitis B virus core antigen

Abstract

References

Citing Literature

References

Information

About Wiley Online Library

Help & Support

Opportunities

Connect with Wiley

Enhancement of cytotoxic T lymphocyte activity by dendritic cells loaded with Tat-protein transduction domain-fused hepatitis B virus core antigen

Abstract

References

Citing Literature

References

Related

Information