Skip to main content
Top
Published in: Journal of Experimental & Clinical Cancer Research 1/2009

Open Access 01-12-2009 | Research

Generation of fusion protein EGFRvIII-HBcAg and its anti-tumor effect in vivo

Authors: Xiao-yi Duan, Dong-gang Han, Ming-xin Zhang, Jian-sheng Wang

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2009

Login to get access

Abstract

The epidermal growth factor receptor variant III (EGFRvIII) is the most common variation of EGFR. Because it shows a high frequency in several different types of tumor and has not been detected in normal tissues, it is an ideal target for tumor specific therapy. In this study, we prepared EGFRvIII-HBcAg fusion protein. After immunization with fusion protein, HBcAg or PBS, the titers of antibody in BALB/c mice immunized with fusion protein reached 2.75 × 105. Western blot analysis demonstrated that the fusion protein had specific antigenicity against anti-EGFRvIII antibody. Further observation showed fusion protein induced a high frequency of IFN-γ-secreting lymphocytes. CD4+T cells rather than CD8+T cells were associated with the production of IFN-γ. Using Renca-vIII(+) cell as specific stimulator, we observed remarkable cytotoxic activity in splenocytes from mice immunized with fusion protein. Mice were challenged with Renca-vIII(+) cells after five times immunization. In fusion protein group, three of ten mice failed to develop tumor and all survived at the end of the research. The weight of tumors in fusion protein were obviously lighter than that in other two groups (t = 4.73, P = 0.044;t = 6.89, P = 0.040). These findings demonstrated that EGFRvIII-HBcAg fusion protein triggered protective responses against tumor expressing EGFRvIII.
Appendix
Available only for authorised users
Literature
1.
go back to reference Jorissen RN, Walker F, Pouliot N, Garrett TP, Ward CW, Burgess AW: Epidermal growth factor receptor: mechanisms of activation and signaling. Exp Cell Res. 2003, 284: 31-53. 10.1016/S0014-4827(02)00098-8.CrossRef Jorissen RN, Walker F, Pouliot N, Garrett TP, Ward CW, Burgess AW: Epidermal growth factor receptor: mechanisms of activation and signaling. Exp Cell Res. 2003, 284: 31-53. 10.1016/S0014-4827(02)00098-8.CrossRef
2.
go back to reference Holbro T, Civenni G, Hynes NE: The ErbB receptors and their role in cancer progression. Exp Cell Res. 2003, 284: 99-110. 10.1016/S0014-4827(02)00099-X.CrossRef Holbro T, Civenni G, Hynes NE: The ErbB receptors and their role in cancer progression. Exp Cell Res. 2003, 284: 99-110. 10.1016/S0014-4827(02)00099-X.CrossRef
3.
go back to reference Herbst RS: Review of epidermal growth factor receptor biology. Int J Radiat Oncol Biol Phys. 2004, 59: 21-26. 10.1016/j.ijrobp.2003.10.027.CrossRef Herbst RS: Review of epidermal growth factor receptor biology. Int J Radiat Oncol Biol Phys. 2004, 59: 21-26. 10.1016/j.ijrobp.2003.10.027.CrossRef
4.
go back to reference Moscatello DK, Holgado-Madruga M, Emlet DR, Montgomery RB, Wong AJ: Constitutive activation of phosphatidylinositol 3-kinase by a naturally occurring mutant epidermal growth factor receptor. J Biol Chem. 1998, 273: 200-206. 10.1074/jbc.273.1.200.CrossRef Moscatello DK, Holgado-Madruga M, Emlet DR, Montgomery RB, Wong AJ: Constitutive activation of phosphatidylinositol 3-kinase by a naturally occurring mutant epidermal growth factor receptor. J Biol Chem. 1998, 273: 200-206. 10.1074/jbc.273.1.200.CrossRef
5.
go back to reference Moscatello DK, Holgado-Madruga M, Godwin AK, Ramirez G, Gunn G, Zoltick PW, et al: Frequent expression of a mutant epidermal growth factor receptor in multiple human tumors. Cancer Res. 1995, 55: 5536-5539. Moscatello DK, Holgado-Madruga M, Godwin AK, Ramirez G, Gunn G, Zoltick PW, et al: Frequent expression of a mutant epidermal growth factor receptor in multiple human tumors. Cancer Res. 1995, 55: 5536-5539.
6.
go back to reference Pedersen MW, Meltorn M, Damstrup L, Poulsen HS: The type III epidermal growth factor receptor mutation. Biological significance and potential target for anti-cancer therapy. Ann Oncol. 2001, 12: 745-760. 10.1023/A:1011177318162.CrossRef Pedersen MW, Meltorn M, Damstrup L, Poulsen HS: The type III epidermal growth factor receptor mutation. Biological significance and potential target for anti-cancer therapy. Ann Oncol. 2001, 12: 745-760. 10.1023/A:1011177318162.CrossRef
7.
go back to reference Pumpens P, Borisova GP, Crowther RA, Grens E: Hepatitis B virus particles as epitope carriers. Intervirol. 1995, 38: 63-74. Pumpens P, Borisova GP, Crowther RA, Grens E: Hepatitis B virus particles as epitope carriers. Intervirol. 1995, 38: 63-74.
8.
go back to reference Karpenko LI, Il'ichev AA: Chimeric hepatitis B core particles as a presentation system of epitopes of foreign proteins. Vestn Russ Akad Med. 1998, 3: 6-9. Karpenko LI, Il'ichev AA: Chimeric hepatitis B core particles as a presentation system of epitopes of foreign proteins. Vestn Russ Akad Med. 1998, 3: 6-9.
9.
go back to reference Moscatello DK, Ramirez G, Wong AJ: A naturally occurring mutant human epidermal growth factor receptor as a target for peptide vaccine immunotherapy of tumors. Cancer, Res. 1997, 57: 1419-1424. Moscatello DK, Ramirez G, Wong AJ: A naturally occurring mutant human epidermal growth factor receptor as a target for peptide vaccine immunotherapy of tumors. Cancer, Res. 1997, 57: 1419-1424.
10.
go back to reference Duan XY, Wang JS, Guo YM, Peng W: Establishment of tumor cell line expressing EGFRvIII gene, U.S.Chinese. J Lymph Onco. 2006, 5: 103-106. Duan XY, Wang JS, Guo YM, Peng W: Establishment of tumor cell line expressing EGFRvIII gene, U.S.Chinese. J Lymph Onco. 2006, 5: 103-106.
11.
go back to reference Luwor RB, Zhu HJ, Walker F, Vitali AA, Perera RM, Burgess AW, et al: The tumor-specific de2-7 epidermal growth factor receptor (EGFR) promotes cells survival and heterodimerizes with the wild-type EGFR. Oncogene. 2004, 23: 6095-6104. 10.1038/sj.onc.1207870.CrossRef Luwor RB, Zhu HJ, Walker F, Vitali AA, Perera RM, Burgess AW, et al: The tumor-specific de2-7 epidermal growth factor receptor (EGFR) promotes cells survival and heterodimerizes with the wild-type EGFR. Oncogene. 2004, 23: 6095-6104. 10.1038/sj.onc.1207870.CrossRef
12.
go back to reference Pedersen MW, Tkach V, Pedersen N, Berezin V, Poulsen HS: Expression of a naturally occurring constitutively active variant of the epidermal growth factor receptor in mouse fibroblasts increases motility. Int J Cancer. 2004, 108: 643-653. 10.1002/ijc.11566.CrossRef Pedersen MW, Tkach V, Pedersen N, Berezin V, Poulsen HS: Expression of a naturally occurring constitutively active variant of the epidermal growth factor receptor in mouse fibroblasts increases motility. Int J Cancer. 2004, 108: 643-653. 10.1002/ijc.11566.CrossRef
13.
go back to reference Boockvar JA, Kapitonov D, Kapoor G, Schouten J, Counelis GJ, Bogler O, et al: Constitutive EGFR signaling confers a motile phenotype to neural stem cells. Mol Cell Neurosci. 2003, 24: 1116-1130. 10.1016/j.mcn.2003.09.011.CrossRef Boockvar JA, Kapitonov D, Kapoor G, Schouten J, Counelis GJ, Bogler O, et al: Constitutive EGFR signaling confers a motile phenotype to neural stem cells. Mol Cell Neurosci. 2003, 24: 1116-1130. 10.1016/j.mcn.2003.09.011.CrossRef
14.
go back to reference Ning Y, Zeineldin R, Liu Y, Rosenberg M, Stack MS, Hudson LG: Down-regulation of integrin alpha2 surface expression by mutant epidermal growth factor receptor (EGFRvIII) induces aberrant cell spreading and focal adhesion formation. Cancer Res. 2005, 65: 9280-9286. 10.1158/0008-5472.CAN-05-0407.CrossRef Ning Y, Zeineldin R, Liu Y, Rosenberg M, Stack MS, Hudson LG: Down-regulation of integrin alpha2 surface expression by mutant epidermal growth factor receptor (EGFRvIII) induces aberrant cell spreading and focal adhesion formation. Cancer Res. 2005, 65: 9280-9286. 10.1158/0008-5472.CAN-05-0407.CrossRef
15.
go back to reference Luwor RB, Johns TG, Murone C, Huang HJ, Cavenee WK, Ritter G, et al: Monoclonal antibody 806 inhibits the growth of tumor xenografts expressing either the de2-7 or amplified epidermal growth factor receptor (EGFR) but not wild-type EGFR. Cancer Res. 2001, 61: 5355-5361. Luwor RB, Johns TG, Murone C, Huang HJ, Cavenee WK, Ritter G, et al: Monoclonal antibody 806 inhibits the growth of tumor xenografts expressing either the de2-7 or amplified epidermal growth factor receptor (EGFR) but not wild-type EGFR. Cancer Res. 2001, 61: 5355-5361.
16.
go back to reference Perera RM, Narita Y, Furnari FB, Gan HK, Murone C, Ahlkvist M, et al: Treatment of human tumor xenografts with monoclonal antibody 806 in combination with a prototypical epidermal growth factor receptor-specific antibody generates enhanced antitumor activity. Clin Cancer Res. 2005, 11: 6390-6399. 10.1158/1078-0432.CCR-04-2653.CrossRef Perera RM, Narita Y, Furnari FB, Gan HK, Murone C, Ahlkvist M, et al: Treatment of human tumor xenografts with monoclonal antibody 806 in combination with a prototypical epidermal growth factor receptor-specific antibody generates enhanced antitumor activity. Clin Cancer Res. 2005, 11: 6390-6399. 10.1158/1078-0432.CCR-04-2653.CrossRef
17.
go back to reference Heimberger AB, Crotty LE, Archer GE, Hess KR, Wiskstrand CJ, Friedman AH, et al: Epidermal growth factor receptor VIII peptide vaccination is efficacious against established intracerebral tumors. Clin Cancer Res. 2003, 9: 4247-4254. Heimberger AB, Crotty LE, Archer GE, Hess KR, Wiskstrand CJ, Friedman AH, et al: Epidermal growth factor receptor VIII peptide vaccination is efficacious against established intracerebral tumors. Clin Cancer Res. 2003, 9: 4247-4254.
18.
go back to reference Wu AH, Xiao J, Anker L, Hall WA, Gregerson DS, Cavenee WK, et al: Identification of EGFRvIII-derived CTL epitopes restricted by HLA A0201 for dendritic cell based immunotherapy of gliomas. J Neuro Oncol. 2006, 76: 23-30. 10.1007/s11060-005-3280-7.CrossRef Wu AH, Xiao J, Anker L, Hall WA, Gregerson DS, Cavenee WK, et al: Identification of EGFRvIII-derived CTL epitopes restricted by HLA A0201 for dendritic cell based immunotherapy of gliomas. J Neuro Oncol. 2006, 76: 23-30. 10.1007/s11060-005-3280-7.CrossRef
19.
go back to reference Broeke LT, Daschbach E, Thomas EK, Andringa G, Berzofsky JA: Dendritic cell-induced activation of adaptive and innate antitumor immunity. J Immunol. 2003, 171: 5842-5852.CrossRef Broeke LT, Daschbach E, Thomas EK, Andringa G, Berzofsky JA: Dendritic cell-induced activation of adaptive and innate antitumor immunity. J Immunol. 2003, 171: 5842-5852.CrossRef
20.
go back to reference Qin Z, Blankenstein T: CD4+ T cell-mediated tumor rejection involves inhibition of angiogenesis that is dependent on IFN-γ receptor expression by nonhematopoietic cells. Immunity. 2000, 12: 677-686. 10.1016/S1074-7613(00)80218-6.CrossRef Qin Z, Blankenstein T: CD4+ T cell-mediated tumor rejection involves inhibition of angiogenesis that is dependent on IFN-γ receptor expression by nonhematopoietic cells. Immunity. 2000, 12: 677-686. 10.1016/S1074-7613(00)80218-6.CrossRef
21.
go back to reference Turley EA, Noble PW, Bourguignon LY: Signaling properties of hyaluronan receptors. J Biol Chem. 2002, 277: 4589-4592. 10.1074/jbc.R100038200.CrossRef Turley EA, Noble PW, Bourguignon LY: Signaling properties of hyaluronan receptors. J Biol Chem. 2002, 277: 4589-4592. 10.1074/jbc.R100038200.CrossRef
22.
go back to reference Patel D, Lahiji A, Patel S, Franklin M, Jimenez X, Hicklin DJ, et al: Monoclonal antibody cetuximab binds to and down-regulates constitutively activated epidermal growth factor receptor vIII on the cell surface. Anticancer Res. 2007, 27: 3355-3366. Patel D, Lahiji A, Patel S, Franklin M, Jimenez X, Hicklin DJ, et al: Monoclonal antibody cetuximab binds to and down-regulates constitutively activated epidermal growth factor receptor vIII on the cell surface. Anticancer Res. 2007, 27: 3355-3366.
Metadata
Title
Generation of fusion protein EGFRvIII-HBcAg and its anti-tumor effect in vivo
Authors
Xiao-yi Duan
Dong-gang Han
Ming-xin Zhang
Jian-sheng Wang
Publication date
01-12-2009
Publisher
BioMed Central
Published in
Journal of Experimental & Clinical Cancer Research / Issue 1/2009
Electronic ISSN: 1756-9966
DOI
https://doi.org/10.1186/1756-9966-28-133

Other articles of this Issue 1/2009

Journal of Experimental & Clinical Cancer Research 1/2009 Go to the issue
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine