Construction and protection effects of the dual expression DNA vaccine pCMV-DKK1/IL10

Abstract

Abstract: AIM: The eukaryotic expression vector of human Dickkopf 1(DKK1) and interleukin-10 (IL-10) DNA vaccine was constructed to evaluate its protection effects in collagen-induced arthritis (CIA) mice. METHODS: Four DNA fragments of human DKK1 with high immunogenicity were selected and then connected with T-cell epitopes. The recombinant sequences of DKK1 and IL-10 were cloned into eukaryotic vector pCMV6-XL5, resulting in the dual expression vector pCMV-DKK1/IL10. The plasmid was transfected into COS7 cells and administrated into the skeletal muscles of BALB/c mice. The expression of the target proteins in COS7 cells and the injection site were detected using Western blotting and immunohistochemistry. The specific antibodies of DNA vaccine was determined using indirect enzyme-linked immunosorbent assay (ELISA). The arthritis incidence, clinical score, radiography and histopathology of the collagen-induced arthritis mice were used as indexes to evaluate the protective effects of the DNA vaccine. RESULTS: The recombinant plasmids pCMV-DKK1/IL10 was successfully constructed and expressed the target proteins both in COS7 cells and in the skeletal muscles of mice. The high expression of anti-human DKK1 antibodies were elicited at the fourth week after immunization with pCMV-DKK1/IL10. In addition, the DNA vaccine attenuated the inflammation and bone erosion in CIA mice.CONCLUSION: The results showed that the immunization of the mice with pCMV-DKK1/IL10 ameliorated the symptoms of CIA mice. This study may provide a potential therapeutic strategy in rheumatology arthritis (RA).

Key words: rheumatoid arthritis(RA), DNA vaccine, DKK1, IL-10

CLC Number:

R965.2

ZHANG Xiao-qing, LIU Si-bo, DOU Yun-peng, DU Yu-xuan, YUAN Hui-hui, ZHAO Wen-ming. Construction and protection effects of the dual expression DNA vaccine pCMV-DKK1/IL10[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(11): 1211-1217.

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