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Clinical and Vaccine Immunology, July 2009, p. 1033-1039, Vol. 16, No. 7
1071-412X/09/$08.00+0 doi:10.1128/CVI.00046-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Specific Antibodies Elicited by a Novel DNA Vaccine Targeting Gastrin-Releasing Peptide Inhibit Murine Melanoma Growth In Vivo
Jing Fang,1,
Yong Lu,2,
Kedong Ouyang,2
Guojun Wu,2
Huiyong Zhang,2,3
Yanhua Liu,2
Yingying Chen,2
Ming Lin,2
Huaqian Wang,2
Liang Jin,2
Rongyue Cao,2
Rouel S. Roque,4
Li Zong,2
Jingjing Liu,2* and
Taiming Li2*
Institute of Chemical Industry of Forest Products, CAF, Nanjing 210042, China,1 Minigene Pharmacy Laboratory, China Pharmaceutical University, Tongjiaxiang 24, Nanjing 210009, China,2 Department of Life Sciences and Technology, Xinxiang Medical University, Xinxiang 453003, China,3 Department of Basic Sciences, Touro University Nevada, Henderson, Nevada 891044
Received 2 February 2009/ Returned for modification 20 April 2009/ Accepted 7 May 2009
The elevated expression and receptor binding of gastrin-releasing peptide (GRP) in various types of cancer, especially in malignant melanoma of the skin, suggest that GRP might be a putative target for immunotherapy in neoplastic diseases. We have therefore constructed a novel DNA vaccine coding for six tandem repeats of a fragment of GRP from amino acids 18 to 27 (GRP6) flanked by helper T-cell epitopes for increased immunogenicity, including HSP65, a tetanus toxoid fragment from amino acids 830 to 844 (T), pan-HLA-DR-binding epitope (PADRE) (P), and two repeats of a mycobacterial HSP70 fragment from amino acids 407 to 426 (M). The anti-GRP DNA vaccine (pCR3.1-VS-HSP65-TP-GRP6-M2) was constructed on a backbone of a pCR3.1 plasmid vector with eight 5'-GACGTT-3' CpG motifs and the VEGF183 signal peptide (VS). Intramuscular (IM) injections of anti-GRP vaccine in mice stimulated the production of high titers of specific antibodies against GRP and suppressed the growth of subcutaneous tumors of B16-F10 melanoma cells. Parallel results were obtained in vitro, showing inhibition of B16-F10 cell proliferation by GRP antisera. IM injections of the DNA vaccine also significantly attenuated tumor-induced angiogenesis associated with intradermal tumors of B16-F10 cells. In addition, lung invasion of intravenously injected cells was highly diminished, suggesting potent antimetastatic activity of the DNA vaccine. These findings support the highly immunogenic and potent antitumorigenic activity of specific anti-GRP antibodies elicited by the anti-GRP DNA vaccine.
* Corresponding author. Mailing address: Minigene Pharmacy Laboratory, Biopharmaceutical College, China Pharmaceutical University, 24 TongjiaXiang, Nanjing 210009, China. Phone for Jingjing Liu: 86-25-83271242. Fax: 86-25-83204240. E-mail: minigene1{at}yahoo.com.cn. Phone for Taiming Li: 86-25-83271369. Fax: 86-25-83204240. E-mail: minigene2{at}yahoo.com.cn
Published ahead of print on 20 May 2009.
Jing Fang and Yong Lu contributed equally to this paper.
Clinical and Vaccine Immunology, July 2009, p. 1033-1039, Vol. 16, No. 7
1071-412X/09/$08.00+0 doi:10.1128/CVI.00046-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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