Reversibility and safety of KISS1 metastasis suppressor gene vaccine in immunocastration of ram lambs

  • Han, Yan-Guo ;
  • Liu, Gui-Qiong ;
  • Jiang, Xun-Ping ;
  • Xiang, Xing-Long ;
  • Huang, Yong-Fu ;
  • Nie, Bin ;
  • Zhao, Jia-Yu ;
  • Nabeel, Ijaz ;
  • Tesema, Birhanu
  • Received : 2017.08.23
  • Accepted : 2017.12.11
  • Published : 2018.06.01


Objective: The aim of this study was to investigate the reversibility and safety of KISS1 metastasis suppressor (KISS1) gene vaccine in immunocastration. Methods: Six eight-week old ram lambs were randomly divided into vaccinated and control groups. The vaccine (1 mg/ram lamb) was injected at weeks 0, 3, and 6 of the study. Blood samples were collected from the jugular vein before primary immunization and at weeks 2, 4, 6, 10, 14, 22, and 30 after primary immunization. All ram lambs were slaughtered at 38 weeks of age, and samples were collected. Results: The specific anti-KISS1 antibody titers in vaccinated animals were significantly higher and the serum testosterone level was significantly lower than those in the control groups from week 4 to 14 after primary immunization (p<0.05). No significant difference was observed at weeks 22 and 30 after the primary immunization. Similar results were also found for scrotal circumference, testicular weight, length, breadth, and spermatogenesis in seminiferous tubules in week 30 after primary immunization. KS (KISS1-hepatitis B surface antigen S) fusion fragment of KISS1 gene vaccine was not detected in host cell genomic DNA of 9 tissues of the vaccinated ram lambs by polymerase chain reaction. Conclusion: The effects of KISS1 gene vaccine in immunocastration were reversible and no integration events were recorded.


KISS1 Metastasis Suppressor;Gene Vaccine;Immunocastration;Reversibility;Safety;Ram Lambs


  1. Han X, Zhou Y, Zeng Y, et al. Effects of active immunization against GnRH versus surgical castration on hypothalamicpituitary function in boars. Theriogenology 2017;97:89-97.
  2. Perez-Linares C, Bolado-Sarabia L, Figueroa-Saavedra F, et al. Effect of immunocastration with Bopriva on carcass characteristics and meat quality of feedlot Holstein bulls. Meat Sci 2017;123:45-9.
  3. Eisemann JD, O'Hare JR, Fagerstone KA. State-level approaches to managing the use of contraceptives in wildlife in the United States. J Zoo Wildl Med 2013;44:S47-51.
  4. Gupta SK, Minhas V. Wildlife population management: are contraceptive vaccines a feasible proposition? Front Biosci (Schol Ed) 2017;9:357-74.
  5. Goericke-Pesch S, Wehrend A, Georgiev P. Suppression of fertility in adult cats. Reprod Domest Anim 2014;49(Suppl 2):33-40.
  6. Messager S, Chatzidaki EE, Ma D, et al. Kisspeptin directly stimulates gonadotropin-releasing hormone release via G protein-coupled receptor 54. Proc Natl Acad Sci USA 2005; 102:1761-6.
  7. Skorupskaite K, George JT, Anderson RA. The kisspeptin- GnRH pathway in human reproductive health and disease. Hum Reprod Update 2014;20:485-500.
  8. Han Y, Liu G, Jiang X, et al. KISS1 can be used as a novel target for developing a DNA immunocastration vaccine in ram lambs. Vaccine 2015;33:777-82.
  9. Martin T, Parker SE, Hedstrom R, et al. Plasmid DNA malaria vaccine: the potential for genomic integration after intramuscular injection. Hum Gene Ther 1999;10:759-68.
  10. Zhao Y, Bao Y, Zhang L, et al. Biosafety of the plasmid pcDNA3- 1E of Eimeria acervulina in chicken. Exp Parasitol 2013;133: 231-6.
  11. Han L, Zhen YH, Liang AX, et al. Oral vaccination with inhibin DNA delivered using attenuated Salmonella choleraesuis for improving reproductive traits in mice. J Basic Microbiol 2014; 54:962-8.
  12. Graham FL, van der Eb AJ. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 1973;52: 456-67.
  13. Wigler M, Pellicer A, Silverstein S, et al. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci USA 1979;76:1373-6.
  14. Zhang F, Fang F, Chang H, et al. Comparison of protection against H5N1 influenza virus in mouse offspring provided by maternal vaccination with HA DNA and inactivated vaccine. Arch Virol 2013;158:1253-65.
  15. Liang A, Riaz H, Dong F, et al. Evaluation of efficacy, biodistribution and safety of antibiotic-free plasmid encoding somatostatin genes delivered by attenuated Salmonella enterica serovar Choleraesuis. Vaccine 2014;32:1368-74.
  16. Huang LY, Wang KY, Xiao D, et al. Safety and immunogenicity of an oral DNA vaccine encoding Sip of Streptococcus agalac tiae from Nile tilapia Oreochromis niloticus delivered by live attenuated Salmonella typhimurium. Fish Shellfish Immunol 2014;38:34-41.
  17. Khan MAH, Ogita K, Ferro VA, et al. Immunisation with a plasmid DNA vaccine encoding gonadotrophin releasing hormone (GnRH-I) and T-helper epitopes in saline suppresses rodent fertility. Vaccine 2008;26:1365-74.
  18. Harstine BR, Maquivar M, Helser LA, et al. Effects of dietary energy on sexual maturation and sperm production in Holstein bulls. J Anim Sci 2015;93:2759-66.
  19. Uliana SR, Affonso MH, Camargo EP, Floeter-Winter LM. Leishmania: genus identification based on a specific sequence of the 18S ribosomal RNA sequence. Exp Parasitol 1991;72: 157-63.
  20. Caldara FR, Moi M, Dos Santos LS, et al. Carcass characteristics and qualitative attributes of pork from immunocastrated animals. Asian-Australas J Anim Sci 2013;26:1630-6.
  21. Moore KL, Mullan BP, Dunshea FR. Boar taint, meat quality and fail rate in entire male pigs and male pigs immunized against gonadotrophin releasing factor as related to body weight and feeding regime. Meat Sci 2017;125:95-101.
  22. Siel D, Vidal S, Sevilla R, et al. Effectiveness of an immunocastration vaccine formulation to reduce the gonadal function in female and male mice by Th1/Th2 immune response. Theriogenology 2016;86:1589-98.
  23. Khan MAH, Ferro VA, Koyama S, et al. Immunisation of male mice with a plasmid DNA vaccine encoding gonadotrophin releasing hormone (GnRH-I) and T-helper epitopes suppresses fertility in vivo. Vaccine 2007;25:3544-53.
  24. Yeung VP, Gieni RS, Umetsu DT, DeKruyff RH. Heat-killed Listeria monocytogenes as an adjuvant converts established murine Th2-dominated immune responses into Th1-dominated responses. J Immunol 1998;161: 4146-52.
  25. Vernot JP, Perez-Quintero LA, Perdomo-Arciniegas AM, Quijano S, Patarroyo ME. Herpesvirus saimiri immortalization of Aotus T lymphocytes specific for an immunogenically modified peptide of Plasmodium falciparum merozoite surface antigen 2. Immunol Cell Biol 2005;83:67-74.
  26. Guo L, Chen J, Qiu Y, et al. Enhanced transcutaneous immunization via dissolving microneedle array loaded with liposome encapsulated antigen and adjuvant. Int J Pharm 2013;447:22-30.
  27. Qiu Y, Guo L, Zhang S, et al. DNA-based vaccination against hepatitis B virus using dissolving microneedle arrays adjuvanted by cationic liposomes and CpG ODN. Drug Deliv 2016;23:2391-8.
  28. Dowsett KF, Tshewang U, Knott LM, Jackson AE, Trigg TE. Immunocastration of colts and immunospeying of fillies. Immunol Cell Biol 1993;71(Pt 5):501-8.
  29. Schulman M, Botha AE, Muenscher SB, et al. Reversibility of the effects of GnRH-vaccination used to suppress reproductive function in mares. Equine Vet J 2013;45:111-3.
  30. Lugar DW, Rhoads ML, Clark-Deener SG, et al. Immunological castration temporarily reduces testis size and function without long-term effects on libido and sperm quality in boars. Animal 2017;11:643-9.
  31. Wolff JA, Ludtke JJ, Acsadi G, Williams P, Jani A. Long-term persistence of plasmid DNA and foreign gene expression in mouse muscle. Hum Mol Genet 1992;1:363-9.


Supported by : Central Universities, National Natural Science Foundation of China