Science in China Series C: Life Sciences

, Volume 51, Issue 4, pp 308–316 | Cite as

Construction of sperm-specific lactate dehydrogenase DNA vaccine and experimental study of its immunocontraceptive effect on mice

  • Yong Chen
  • Duo Zhang
  • Na Xin
  • YongZhong Xiong
  • Ping Chen
  • Bo Li
  • XiangDong Tu
  • FengHua LanEmail author


Lactate dehydrogenase C4 (LDHC4) is a key enzyme for sperm metabolism. It is distributed specifically in testis and is highly immunogenic. In this study, two DNA vaccines pVAX1-hLDHC and pVAX1-mLDHC were constructed by inserting coding sequences of human and mice LDHC4 into the eukaryotic expression vector pVAX1. The production of LDHC4 specific antibodies was induced in the sera of vaccinated mice and the reproductive tract secretions of vaccinated female mice through immunization by mucosal surface instillation. Furthermore, the antibody titer increased with the times of immunization. In the mating experiment, the number of newborns of the vaccinated mice reduced significantly and some immunized female mice even lost the ability to bear any offsprings, suggesting that the difference between the immunized and control mice was statistically significant. Sperm agglutination analysis indicated that both the antisera from immunized mice and the reproductive tract secretions of vaccinated female mice could agglutinate normal sperms. Results of immunohistochemistry showed that the antibodies present in the sera of immunized mice and the reproductive tract secretions of vaccinated female mice could specifically react with LDHC4 antigen, which mainly locates in the cytoplasm, acrosome membrane externa and acrosome capsule of the sperm. Taken together, our results indicated that the constructed contraceptive DNA vaccines did yield immunocontraceptive effects on mice and this would enable clinical trials in near future.


sperm-specific lactate dehydrogenase DNA vaccine contraceptive vaccine mucosal immunization 


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  1. 1.
    Naz R K. Vaccine for contraception targeting sperm. Immunol Rev, 1999, 171(10): 193–202, 10582172, 10.1111/j.1600-065X.1999.tb01349.x, 1:CAS:528:DyaK1MXnvVSiu7k%3DPubMedCrossRefGoogle Scholar
  2. 2.
    Shelton J A, Goldberg E. Local reproductive tract immunity to sperm-specific lactate dehydrogenase-C4. Biol Reprod, 1986, 35(4): 873–876, 3814701, 10.1095/biolreprod35.4.873, 1:CAS:528:DyaL2sXkvVelsw%3D%3DPubMedCrossRefGoogle Scholar
  3. 3.
    Mahi-Brown C A, van de Voort C A, McGuinness R P, et al. Immunization of male but not female mice with the sperm-specific isozyme of lactate dehydrogenase (LDH-C4) impairs fertilization in vivo. Am J Reprod Immunol, 1990, 24(1): 1–8, 2285453, 1:STN:280:By6C3snmvVM%3DPubMedCrossRefGoogle Scholar
  4. 4.
    Gupta G S, Syal N. Immune responses of chemically modified homologous LDH-C4 and their effect on fertility regulation in mice. Am J Reprod Immunol, 1997, 37(2): 206–211, 9083619, 1:STN:280:ByiB3sjhsVU%3DPubMedCrossRefGoogle Scholar
  5. 5.
    Goldberg E. Infertility in female rabbits immunized with lactate dehydrogenase X. Science, 1973, 181(98): 458–459, 4718114, 10.1126/science.181.4098.458, 1:CAS:528:DyaE3sXltVenu7g%3DPubMedCrossRefGoogle Scholar
  6. 6.
    Gupta G S, Malhotra R, Sehgal S. Regulation of fertility in female mice after immunization with human sperm specific LDH: Role in conception and contraception. Indian J Exp Biol, 1994, 32(1): 14–19, 8045599, 1:CAS:528:DyaK2cXps1Srsw%3D%3DPubMedGoogle Scholar
  7. 7.
    Wheat T E, Goldberg E. Sperm-specific lactate dehydrogenase C4: Antigenic structure and immunosuppression of fertility. Isozymes Curr Top Biol Med Res, 1983, 7113–7130Google Scholar
  8. 8.
    Shen H, Goldberg E, Saltzman W M. Gene expression and mucosal immune responses after vaginal DNA immunization in mice using a controlled delivery matrix. J Control Release, 2003, 86(2–3): 339–348, 12526829, 10.1016/S0168-3659(02)00354-1, 1:CAS:528:DC%2BD3sXms1ClPubMedCrossRefGoogle Scholar
  9. 9.
    Shi S Q, Wang J L, Peng J P, et al. Oral feeding and nasal instillation immunization with Microtus brandti lactate dehydrogenase C epitope DNA vaccine reduces fertility in mice via specific antibody responses. Fert Steril, 2005, 84(3): 781–784, 10.1016/j.fertnstert.2005.03.060, 1:CAS:528:DC%2BD2MXhtFeisLzMCrossRefGoogle Scholar
  10. 10.
    Chang J J, Peng J P, Yang Y, et al. Study on the antifertility effects of the plasmid DNA vaccine expressing partial brLDH-C4′. Reproduction, 2006, 131: 183–192, 16388021, 10.1530/rep.1.00262, 1:CAS:528:DC%2BD28XhsFegs7w%3DPubMedCrossRefGoogle Scholar
  11. 11.
    Philippopoulos M, Xiang Y, Lim C. Identifying the mechanism of protein loop closure: A molecular dynamics simulation of the Bacillus stearothermophilus LDH loop in solution. Prot Eng, 1995, 8(6): 565–573, 10.1093/protein/8.6.565, 1:CAS:528:DyaK2MXotVSqtbs%3DCrossRefGoogle Scholar
  12. 12.
    Bradley M P, Geelan A, Leitch V, et al. Cloning, sequencing, and characterization of LDH-C4 from a fox testis cDNA library. Mol Reprod Dev, 1996, 44(4): 452–459, 8844687, 10.1002/(SICI)1098-2795(199608)44:4<452::AID-MRD4>3.0.CO;2-K, 1:CAS:528:DyaK28XkvFGrtL8%3DPubMedCrossRefGoogle Scholar
  13. 13.
    Goldberg E, VandeBerg J L, Mahony M C, et al. Immune response of male baboons to testis-specific LDH-C(4). Contraception, 2001, 64(2): 93–98, 11704085, 10.1016/S0010-7824(01)00227-X, 1:CAS:528:DC%2BD3MXotFSqsLw%3DPubMedCrossRefGoogle Scholar
  14. 14.
    Klavinskis L S, Barnfield C, Gao L, et al. Intranasal immunization with plasmid DNA-lipid complexes elicits mucosal immunity in the female genital and rectal tracts. J Immunol, 1999, 162(1): 254–262, 9886393, 1:CAS:528:DyaK1MXjt1Ogtw%3D%3DPubMedGoogle Scholar
  15. 15.
    Kuklin N A, Rott L, Feng N, et al. Protective intestinal anti-rotavirus B cell immunity is dependent on α4β7 integrin expression but does not require IgA antibody production. J Immunol, 2001, 166(3): 1894–1902, 11160237, 1:CAS:528:DC%2BD3MXhtVSis7w%3DPubMedCrossRefGoogle Scholar
  16. 16.
    Kanellos T S, Byarugaba D K, Russell P H, et al. Naked DNA when co-administered intranasally with heat-labile enterotoxin of Escherichia coli primes effectively for systemic B-and T-cell responses to the encoded antigen. Immunol Lett, 2000, 74(3): 215–220, 11064104, 10.1016/S0165-2478(00)00257-1, 1:CAS:528:DC%2BD3cXosV2itrY%3DPubMedCrossRefGoogle Scholar
  17. 17.
    Kaneko H, Bednarek I, Wierzbicki A, et al. Oral DNA vaccination promotes mucosal and systemic immune responses to HIV envelope glycoprotein. Virology, 2000, 267(1): 8–16, 10648178, 10.1006/viro.1999.0093, 1:CAS:528:DC%2BD3cXmsFamtg%3D%3DPubMedCrossRefGoogle Scholar
  18. 18.
    Roy K, Mao H Q, Huang S K, et al. Oral gene delivery with chitosan-DNA nanoparticles generates immunologic protection in a murine model of peanut allergy. Nat Med, 1999, 5(4): 387–391, 10202926, 10.1038/7385, 1:CAS:528:DyaK1MXisFOlsr0%3DPubMedCrossRefGoogle Scholar
  19. 19.
    Lee S, Gierynska M, Eo S K, et al. Influence of DNA encoding cytokines on systemic and mucosal immunity following genetic vaccination against herpes simplex virus. Microbes Infect, 2003, 5(7): 571–578, 12787733, 10.1016/S1286-4579(03)00108-4, 1:CAS:528:DC%2BD3sXkt1Knur0%3DPubMedCrossRefGoogle Scholar
  20. 20.
    Gerdts V, Tsang C, Griebel P J, et al. DNA vaccination in utero: A new approach to induce protective immunity in the newborn. Vaccine, 2004, 22(13–14): 1717–1727, 15068855, 10.1016/j.vaccine.2003.05.004, 1:CAS:528:DC%2BD2cXivVGjs7o%3DPubMedCrossRefGoogle Scholar

Copyright information

© Science Press 2008

Authors and Affiliations

  • Yong Chen
    • 1
  • Duo Zhang
    • 1
  • Na Xin
    • 1
  • YongZhong Xiong
    • 1
  • Ping Chen
    • 1
  • Bo Li
    • 1
  • XiangDong Tu
    • 1
  • FengHua Lan
    • 1
    Email author
  1. 1.Research Laboratory of Medical Genetics, PLA Institute of Technology for Birth Control, Fetal Health and Child GrowthFuzhou General Hospital of Nanjing Military RegionFuzhouChina

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