Yange Niu, Ye Liu, Limin Yang, Hongren Qu, Jingyi Zhao, Rongliang Hu, Jing Li and Wenjun Liu. Immunogenicity of multi-epitope-based vaccine candidates administered with the adjuvant Gp96 against rabies[J]. Virologica Sinica, 2016, 31(2): 168-175. doi: 10.1007/s12250-016-3734-4
Citation: Yange Niu, Ye Liu, Limin Yang, Hongren Qu, Jingyi Zhao, Rongliang Hu, Jing Li, Wenjun Liu. Immunogenicity of multi-epitope-based vaccine candidates administered with the adjuvant Gp96 against rabies .VIROLOGICA SINICA, 2016, 31(2) : 168-175.  http://dx.doi.org/10.1007/s12250-016-3734-4

Immunogenicity of multi-epitope-based vaccine candidates administered with the adjuvant Gp96 against rabies

  • Corresponding author: Jing Li, lj418@163.com Wenjun Liu, liuwj@im.ac.cn
  • Received Date: 29 January 2016
    Accepted Date: 16 March 2016
    Published Date: 06 April 2016
  • Rabies, a zoonotic disease, causes > 55,000 human deaths globally and results in at least 500 million dollars in losses every year. The currently available rabies vaccines are mainly inactivated and attenuated vaccines, which have been linked with clinical diseases in animals. Thus, a rabies vaccine with high safety and efficacy is urgently needed. Peptide vaccines are known for their low cost, simple production procedures and high safety. Therefore, in this study, we examined the efficacy of multi-epitope-based vaccine candidates against rabies virus. The ability of various peptides to induce epitope-specific responses was examined, and the two peptides that possessed the highest antigenicity and conservation, i.e., AR16 and hPAB, were coated with adjuvant canineGp96 and used to prepare vaccines. The peptides were prepared as an emulsion of oil in water (O/W) to create three batches of bivalent vaccine products. The vaccine candidates possessed high safety. Virus neutralizing antibodies were detected on the day 14 after the first immunization in mice and beagles, reaching 5–6 IU/mL in mice and 7–9 IU/mL in beagles by day 28. The protective efficacy of the vaccine candidates was about 70%–80% in mice challenged by a virulent strain of rabies virus. Thus, a novel multi-epitope-based rabies vaccine with Gp96 as an adjuvant was developed and validated in mice and dogs. Our results suggest that synthetic peptides hold promise for the development of novel vaccines against rabies.

  • 加载中
    1. Bachmann MF, Jennings GT. 2010. Vaccine delivery: a matter of size, geometry, kinetics and molecular patterns. Nat Rev Immunol, 10: 787-796.
        doi: 10.1038/nri2868

    2. Bolhassani A, Rafati S. 2008. Heat-shock proteins as powerful weapons in vaccine development. Expert Rev Vaccines, 7: 1185-1199.
        doi: 10.1586/14760584.7.8.1185

    3. Cai K, Feng JN, Wang Q, Li T, Shi J, Hou XJ, Gao X, Liu H, Tu W, Xiao L, Wang H. 2010. Fine mapping and interaction analysis of a linear rabies virus neutralizing epitope. Microbes Infect, 12: 948-955.
        doi: 10.1016/j.micinf.2010.06.005

    4. Chen C, Li J, Bi Y, Jia X, Meng S, Sun L, Liu W. 2012. Gp96 enhances the immunogenicity of subunit vaccine of porcine reproductive and respiratory syndrome virus. Virus Res, 167: 162-172.
        doi: 10.1016/j.virusres.2012.04.011

    5. Cliquet F, Aubert M, Sagne L. 1998. Development of a fluorescent antibody virus neutralisation test (FAVN test) for the quantitation of rabies-neutralising antibody. J Immunol Methods, 212: 79-87.
        doi: 10.1016/S0022-1759(97)00212-3

    6. Commission of Chinese Veterinary Pharmacopoeia. 2010. Veterinary Pharmacopoeia of the People's Republic of China. Chemical Industry Press: 163-168.

    7. Crooks GE, Hon G, Chandonia JM, Brenner SE. 2004. WebLogo: a sequence logo generator. Genome Res, 14: 1188-1190.
        doi: 10.1101/gr.849004

    8. Degen WG, Jansen T, Schijns VE. 2003. Vaccine adjuvant technology: from mechanistic concepts to practical applications. Expert Rev Vaccines, 2: 327-335.
        doi: 10.1586/14760584.2.2.327

    9. Dey AK, Srivastava IK. 2011. Novel adjuvants and delivery systems for enhancing immune responses induced by immunogens. Expert Rev Vaccines, 10: 227-251.
        doi: 10.1586/erv.10.142

    10. Dietzschold B, Gore M, Marchadier D, Niu HS, Bunschoten HM, Otvos L, Jr., Wunner WH, Ertl HC, Osterhaus AD, Koprowski H. 1990. Structural and immunological characterization of a linear virus-neutralizing epitope of the rabies virus glycoprotein and its possible use in a synthetic vaccine. J Virol, 64: 3804-3809.

    11. Dietzschold B, Li J, Faber M, Schnell M. 2008. Concepts in the pathogenesis of rabies. Future Virol, 3: 481-490.
        doi: 10.2217/17460794.3.5.481

    12. Guy B. 2007. The perfect mix: recent progress in adjuvant research. Nat Rev Microbiol, 5: 505-517.

    13. Houimel M, Dellagi K. 2009. Peptide mimotopes of rabies virus glycoprotein with immunogenic activity. Vaccine, 27: 4648-4655.
        doi: 10.1016/j.vaccine.2009.05.055

    14. Kaur M, Garg R, Singh S, Bhatnagar R. 2015. Rabies vaccines: where do we stand, where are we heading? Expert Rev Vaccines, 14: 369-381.
        doi: 10.1586/14760584.2015.973403

    15. Koraka P, Bosch BJ, Cox M, Chubet R, Amerongen G, Lovgren-Bengtsson K, Martina BE, Roose J, Rottier PJ, Osterhaus AD. 2014. A recombinant rabies vaccine expressing the trimeric form of the glycoprotein confers enhanced immunogenicity and protection in outbred mice. Vaccine, 32: 4644-4650.
        doi: 10.1016/j.vaccine.2014.06.058

    16. Kundi M. 2007. New hepatitis B vaccine formulated with an improved adjuvant system. Expert Rev Vaccines, 6: 133-140.
        doi: 10.1586/14760584.6.2.133

    17. Li H, Nookala S, Re F. 2007. Aluminum Hydroxide Adjuvants Activate Caspase-1 and Induce IL-1 and IL-18 Release. J Immunol, 178: 5271-5276.
        doi: 10.4049/jimmunol.178.8.5271

    18. Li Y, Song H, Li J, Wang Y, Yan X, Zhao B, Zhang X, Wang S, Chen L, Qiu B, Meng S. 2011. Hansenula polymorpha expressed heat shock protein gp96 exerts potent T cell activation activity as an adjuvant. J Biotechnol, 151: 343-349.
        doi: 10.1016/j.jbiotec.2010.12.006

    19. Liu X, Yang Y, Sun Z, Chen J, Ai J, Dun C, Fu ZF, Niu X, Guo X. 2014. A recombinant rabies virus encoding two copies of the glycoprotein gene confers protection in dogs against a virulent challenge. PLoS One, 9: e87105.
        doi: 10.1371/journal.pone.0087105

    20. Mansfield KL, Johnson N, Fooks AR. 2004. Identification of a conserved linear epitope at the N terminus of the rabies virus glycoprotein. J Gen Virol, 85: 3279-3283.
        doi: 10.1099/vir.0.80362-0

    21. Marissen WE, Kramer RA, Rice A, Weldon WC, Niezgoda M, Faber M, Slootstra JW, Meloen RH, Clijsters-van der Horst M, Visser TJ, Jongeneelen M, Thijsse S, Throsby M, de Kruif J, Rupprecht CE, Dietzschold B, Goudsmit J, Bakker AB. 2005. Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: fine mapping and escape mutant analysis. J Virol, 79: 4672-4678.
        doi: 10.1128/JVI.79.8.4672-4678.2005

    22. McCluskie MJ, Weeratna RD. 2001. Novel adjuvant systems. Curr Drug Targets Infect Disord, 1: 263-271.
        doi: 10.2174/1568005014605991

    23. Meloen RH, Langeveld JP, Schaaper WM, Slootstra JW. 2001. Synthetic peptide vaccines: unexpected fulfillment of discarded hope? Biologicals, 29: 233-236.
        doi: 10.1006/biol.2001.0298

    24. Ni Y, Tominaga Y, Honda Y, Morimoto K, Sakamoto S, Kawai A. 1995. Mapping and characterization of a sequential epitope on the rabies virus glycoprotein which is recognized by a neutralizing monoclonal antibody, RG719. Microbiol Immunol, 39: 693-702.
        doi: 10.1111/mim.1995.39.issue-9

    25. Niederhauser S, Bruegger D, Zahno ML, Vogt HR, Peterhans E, Zanoni R, Bertoni G. 2008. A synthetic peptide encompassing the G5 antigenic region of the rabies virus induces high avidity but poorly neutralizing antibody in immunized animals. Vaccine, 26: 6749-6753.
        doi: 10.1016/j.vaccine.2008.10.020

    26. Pulmanausahakul R, Li J, Schnell MJ, Dietzschold B. 2008. The glycoprotein and the matrix protein of rabies virus affect pathogenicity by regulating viral replication and facilitating cell-to-cell spread. J Virol, 82: 2330-2338.
        doi: 10.1128/JVI.02327-07

    27. Rupprecht CE, Gibbons RV. 2004. Clinical practice. Prophylaxis against rabies. N Engl J Med, 351: 2626-2635.

    28. Takayama-Ito M, Inoue K, Shoji Y, Inoue S, Iijima T, Sakai T, Kurane I, Morimoto K. 2006. A highly attenuated rabies virus HEP-Flury strain reverts to virulent by single amino acid substitution to arginine at position 333 in glycoprotein. Virus Res, 119: 208-215.
        doi: 10.1016/j.virusres.2006.01.014

    29. Tao X, Han N, Guo Z, Tang Q, Rayner S, Liang G. 2013. Molecular characterization of China human rabies vaccine strains. Virol Sin, 28: 116-123.
        doi: 10.1007/s12250-013-3314-9

    30. Villarreal-Ramos B. 2009. Towards improved understanding of protective mechanisms induced by the BCG vaccine. Expert Rev Vaccines, 8: 1531-1534.
        doi: 10.1586/erv.09.109

    31. Wang S, Qiu L, Liu G, Li Y, Zhang X, Jin W, Gao GF, Kong X, Meng S. 2011. Heat shock protein gp96 enhances humoral and T cell responses, decreases Treg frequency and potentiates the anti-HBV activity in BALB/c and transgenic mice. Vaccine, 29: 6342-6351.
        doi: 10.1016/j.vaccine.2011.05.008

    32. World Organisation for Animal Health. 2013. Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. OIE Terrestrial Manual: Chapter 2. 1. 13.

    33. Xiao XX, Zhang Y, Liu JX, Wei QL, Yin XP. 2015. Immunoenhancement with flagellin as an adjuvant to whole-killed rabies vaccine in mice. Arch Virol, 161: 685-691.

    34. Xue XH, Zheng XX, Wang HL, Ma JZ, Li L, Gai WW, Wang TC, Yang ST, Xia XZ. 2014. An inactivated recombinant rabies CVS-11 virus expressing two copies of the glycoprotein elicits a higher level of neutralizing antibodies and provides better protection in mice. Virus Genes, 48: 411-420.
        doi: 10.1007/s11262-014-1049-9

    35. Yang L, Cen J, Xue Q, Li J, Bi Y, Sun L, Liu W. 2013. Identification of rabies virus mimotopes screened from a phage display peptidelibrary with purified dog anti-rabies virus serum IgG. Virus Res, 174: 47-51.
        doi: 10.1016/j.virusres.2013.02.013

    36. Yuan Z, Zhang S, Liu Y, Zhang F, Fooks AR, Li Q, Hu R. 2008. A recombinant pseudorabies virus expressing rabies virus glycoprotein: safety and immunogenicity in dogs. Vaccine, 26: 1314-1321.
        doi: 10.1016/j.vaccine.2007.12.050

    37. Zhang Y, Zhang S, Li W, Hu Y, Zhao J, Liu F, Lin H, Liu Y, Wang L, Xu S, Hu R, Shao H, Li L. 2016. A novel rabies vaccine based-on toll-like receptor 3 (TLR3) agonist PIKA adjuvant exhibiting excellent safety and efficacy in animal studies. Virology, 489: 165-172.
        doi: 10.1016/j.virol.2015.10.029

  • 加载中

Figures(4) / Tables(2)

Article Metrics

Article views(5514) PDF downloads(23) Cited by(0)

Related
Proportional views
    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Immunogenicity of multi-epitope-based vaccine candidates administered with the adjuvant Gp96 against rabies

      Corresponding author: Jing Li, lj418@163.com
      Corresponding author: Wenjun Liu, liuwj@im.ac.cn
    • 1. Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
    • 2. University of Chinese Academy of Sciences, Beijing 100049, China
    • 3. Laboratory of Epidemiology and Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Veterinary Research Institute, Academy of Military Medical Sciences, Changchun 130122, China
    • 4. Beijing Center for Animal Disease Control and Prevention Chinese Veterinary Medical Association of Animal Clinic Branch, Beijing 102629, China
    • 5. State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China

    Abstract: Rabies, a zoonotic disease, causes > 55,000 human deaths globally and results in at least 500 million dollars in losses every year. The currently available rabies vaccines are mainly inactivated and attenuated vaccines, which have been linked with clinical diseases in animals. Thus, a rabies vaccine with high safety and efficacy is urgently needed. Peptide vaccines are known for their low cost, simple production procedures and high safety. Therefore, in this study, we examined the efficacy of multi-epitope-based vaccine candidates against rabies virus. The ability of various peptides to induce epitope-specific responses was examined, and the two peptides that possessed the highest antigenicity and conservation, i.e., AR16 and hPAB, were coated with adjuvant canineGp96 and used to prepare vaccines. The peptides were prepared as an emulsion of oil in water (O/W) to create three batches of bivalent vaccine products. The vaccine candidates possessed high safety. Virus neutralizing antibodies were detected on the day 14 after the first immunization in mice and beagles, reaching 5–6 IU/mL in mice and 7–9 IU/mL in beagles by day 28. The protective efficacy of the vaccine candidates was about 70%–80% in mice challenged by a virulent strain of rabies virus. Thus, a novel multi-epitope-based rabies vaccine with Gp96 as an adjuvant was developed and validated in mice and dogs. Our results suggest that synthetic peptides hold promise for the development of novel vaccines against rabies.