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Acta Biochimica et Biophysica Sinica 2009 41(2):137-145; doi:10.1093/abbs/gmn016
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© The Author 2009. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

Development of prophylactic recombinant HPV58-attenuated Shigella live vector vaccine and evaluation of its protective efficacy and immunogenicity in the guinea pig keratoconjunctivitis model

Wensheng Li1,2, Hongli Liu3, Xiaofeng Yang2, Jin Zheng2, Yili Wang2 and Lusheng Si2,*

1 Shaanxi Provincial People's Hospital, The Third Affiliated Hospital, Medical College of Xi'an Jiaotong University, Xi'an 710068, China
2 Institute for Cancer Research, Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education, Xi'an Jiaotong University, Xi'an 710061, China
3 The First affiliated Hospital, Medical College of Xi'an Jiaotong University, Xi'an 710061, China

* Correspondence address. Tel: +86-29-82655191; Fax: +86-29-82655499; E-mail: slusheng{at}yahoo.com


  Abstract

To develop a prophylactic recombinant HPV58L1-attenuated Shigella live vector vaccine and evaluate its protective efficacy and immunogenicity in the guinea pig keratoconjunctivitis model, the HPV58L1 gene was cloned into vector pUCmt, and then subcloned into the suicide vector pCVD442. The recombinant plasmid pCVD442-HPV58L1 was introduced into attenuated Shigella (sf301:{Delta}virG) with the helper plasmid PRK2013 by filter mating. The positive colonies were harvested and confirmed by polymerase chain reaction. The expression of the HPV58L1 protein with a molecular weight of 60 kDa was confirmed by western blot. The ability of the interested protein to self-assemble into virus-like particles was identified by transmission electron microscope, and murine erythrocyte hemagglutination assay. The guinea pig keratoconjunctivitis model was used to evaluate the protective efficacy and immunogenicity of the vaccine. Animal experiments showed that there was no keratoconjunctivitis occurred in the immunized group (HPV58-attenuated Shigella), and the serum levels of anti-HPV58L1-IgG and -IgA were obviously increased (P < 0.05), but the anti-sf301 LPS-IgG just slightly increased (P > 0.05). Enzyme-linked immunosorbent spot assay showed that HPV58L1-specific IgA-antibody-secreting cells (ASC) and IgG-ASC of spleen and lymph nodes were also obviously increased (P < 0.01). In this study, a recombinant HPV58L1-attenuated Shigella live vector vaccine was successfully constructed, and it could induce strong humoral immune responses in the immunized animals, and induce protective antibody production.

Keywords    HPV58; Shigella; live vector vaccine; recombination

Received: September 9, 2008; Accepted: November 10, 2008
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