Latest Cover

Online Office

Contact Us

Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
Sponsored by:Sichuan Society of Zoologists; Chengdu Giant Panda Breeding Research Foundation; Sichuan Association of Wildlife Conservation; Sichuan University
Address:College of Life Sciences, Sichuan University, No.29, Wangjiang Road, Chengdu, Sichuan Province, 610064, China
Email:[email protected] & [email protected]
Your Position :Home->Past Journals Catalog->2020 Vol.39 No.3

Evaluation of Contraceptive Effects of Canine Zona Pellucida 3 DNA Vaccine in a Mouse Model
Author of the article:WANG Ying1, ZHANG Fuchun2*
Author's Workplace:1. College of Textile and Fashion Design, Xinjiang University, Urumqi 830046, China;
2. College of Life Science and Technology, Xinjiang University, Urumqi 830046, China
Key Words:canine; zona pellucida 3; DNA vaccine; mouse model; immunocontraception
Abstract:To evaluate the contraceptive effects of the canine zona pellucida 3 (CZP3) DNA vaccine, initial evaluation of the contraceptive efficacy was made in mouse model in this study. Twelve female mice were divided into 2 groups which were intramuscularly immunized with 50 μg of pcDNA3-CZP3 (experiment group) and pcDNA3 plasmids (negative control group), respectively, and 6 electric pulses (30 V) were delivered subsequently. After the first immunization, blood was collected weekly; the antibody levels of CZP3 and antibody titers of CZP3 at the 6th week after the first immunization were detected by ELISA. The mice were mated with male mice at the 6th week after the first immunization, the litter size of each mouse was calculated and the data were analyzed statistically. Ovaries were collected at the 24th week after the first immunization and conducted for HE staining. Finally, the indirect immunofluorescence of mouse and dog zona pellucida were detected by mouse serum. The results indicated that compared with the negative control group: 1) The levels of antibodies against CZP3 were significantly increased at the 2nd week after the first immunization in the experiment group, and lasted for up to 24 weeks. 2) The antibody titer of CZP3 at 1∶4 000 dilution was significantly different. 3) The fertility rate of mice in the experiment group fell from 100% to 50%, and the litter size fell from 6.667±0.422 to 2.500±1.455. However, the correlation between the litter size and antibody levels in the experiment group was negative. 4) HE staining of ovarian sections showed that the immunization with CZP3 DNA vaccine would not influence the development of mouse ovaries. 5) The results of indirect immunofluorescence assay showed that the serum of mouse immunized with CZP3 could react with mouse and dog zona pellucida. Therefore, these results suggested that CZP3 DNA vaccine could reduce the fertility rate and litter size significantly in mouse model without influencing the development of ovaries.
2020,39(3): 266-273 收稿日期:2019-10-16
作者簡介:王穎(1980—),女,講師,E-mail:[email protected]
*通信作者:張富春,E-mail:[email protected]
樊姝彤, 尤新國, 滿怡, 等. 2017. 免疫避孕疫苗的研究進展[J]. 中國男科學雜志, 31(3): 69-72.
李斌, 李瑩瑩, 王瑞, 等. 2016. 呼和浩特市流浪犬狂犬病病毒血清學調查[J]. 黑龍江畜牧獸醫, (22): 208-210.
汪天平, 操治國. 2018. 中國棘球蚴病防控進展及其存在的問題[J]. 中國寄生蟲學與寄生蟲病雜志, 36(3): 291-296.
王金霞. 2016. 犬貓管理與流浪犬貓治理[J]. 中國畜牧業, (20): 64-65.
王穎, 張貝貝, 張冀, 等. 2016. 犬透明帶3(CZP3)DNA疫苗免疫有效降低小鼠生育能力[J]. 細胞與分子免疫學雜志, 32(10): 1331-1335.
張富春, 李江偉, 蘭海燕, 等. 2008. 分子生物學實驗技術[M]. 烏魯木齊: 新疆大學出版社.
莊心依, 周雙雙, 王夢倩, 等. 2017. 多主體合作視角下的城市流浪犬管理研究——基于LA市的實證分析[J]. 畜牧與飼料科學, 38(9): 68-72.
Alarcon JB, Waine GW, McManus DP. 1999. DNA vaccines: technology and application as anti-parasite and antimicrobial agents[J]. Advances in Parasitology, 42: 343-410.
Almeida RR, Raposo RA, Coirada FC, et al. 2015. Modulating APOBEC expression enhances DNA vaccine immunogenicity[J]. Immunology and Cell Biology, 93(10): 868-876.
Bagavant H, Fusi FM, Baisch J, et al. 1997. Immunogenicity and contraceptive potential of human zona pellucida 3 peptide vaccine[J]. Biology of Reproduction, 56(3): 764-770.
Bergamaschi C, Kulkarni V, Rosati M, et al. 2015. Intramuscular delivery of heterodimeric IL-15 DNA in macaques produces systemic levels of bioactive cytokine inducing proliferation of NK and T cells[J]. Gene Therapy, 22(1): 76-86.
Broderick KE,Humeau LM. 2015. Electroporation-enhanced delivery of nucleic acid vaccines[J]. Expert Review of Vaccines, 14(2): 195-204.
Cervantes-Villagrana AR, Hernandez-Pando R, Biragyn A, et al. 2013. Prime-boost BCG vaccination with DNA vaccines based in bata-defensin-2 and mycobacterial antigens ESAT6 or AG85B improve protection in a tuberculosis experimental model[J]. Vaccine, 31(4): 676-684.
Chuang I, Sedegah M, Cicatelli S, et al. 2013. DNA prime/adenovirus boost malaria vaccine encoding P. falciparum CSP and AMA1 induces sterile protection associated with cell-mediated immunity[J/OL]. PLoS ONE, 8(2): e55571 [2019-07-31].
Fossum E, Grodeland G, Terhorst D, et al. 2015. Vaccine molecules targeting Xcr1 on cross-presenting DCs induce protective CD8 T-cell responses against influenza virus[J]. European Journal of Immunology, 45(2): 624-635.
Fournillier A, Frelin L, Jacquier E, et al. 2013. A heterologous prime/boost vaccination strategy enhances the immunogenicity of therapeutic vaccines for hepatitis C virus[J]. The Journal of Infectious Diseases, 208(6): 1008-1019.
Fowler V, Robinson L,Bankowski B, et al. 2012. A DNA vaccination regime including protein boost and electroporation protects cattle against foot-and-mouth disease[J]. Antiviral Research, 94(1): 25-34.
Gupta N, Chakrabarti K, Prakash K, et al. 2013. Immunogenicity and contraceptive efficacy of Escherichia coli-expressed recombinant porcine zona pellucida proteins[J]. American Journal of Reproductive Immunology, 70(2): 139-152.
Gupta SK, Malik A, Arukha AP, et al. 2015. Ovarian and oocyte targets for development of female contraceptives [J]. Expert Opinion on Therapeutic Targets, 19(11): 1433-1446.
Hardy CM, ten Have JFM, Mobbs KJ, et al. 2002. Assessment of the immunocontraceptive effect of a zona pellucida 3 peptide antigen in wild mice[J]. Reproduction Fertility and Development, 14(3): 151-155.
Jackson RJ, Maguire DJ, Hinds LA, et al. 1998. Infertility in mice induced by a recombinant ectromelia virus expressing mouse zona pellucida glycoprotein 3[J]. Biology of Reproduction, 58(1): 152-159.
Kim D, Hung CF, Wu TC, et al. 2010. DNA vaccine with α-galactosyleceramide at prime phase enhances anti-tumor immunity after boosting with antigen-expressing dendritic cells[J]. Vaccine, 28(45): 7297-7305.
Kitchener AL, Harman A, Kay DJ, et al. 2009a. Immunocontraception of eastern grey kangaroos (Macropus giganteus) with recombinant brushtail possum (Trichosurus vulpecula) ZP3 protein[J]. Journal of Reproductive Immunology, 79(2): 156-162.
Kitchener AL, Kay DJ, Walters B, et al. 2009b. The immune response and fertility of koalas (Phascolarctos cinereus) immunised with porcine zonae pellucidae or recombinant brushtail possum ZP3 protein[J]. Journal of Reproductive Immunology, 82(1): 40-47.
Kutzler MA, Weiner DB. 2008. DNA vaccines: ready for prime time?[J]. Nature Reviews Genetics, 9(10): 776-788.
Lee SH, Danishmalik SN, Sin JI. 2015. DNA vaccines, electroporation and their applications in cancer treatment[J]. Human Vaccines & Immunotherapeutics, 11(8): 1889-1900.
Lloyd ML, Shellam GR, Papadimitriou JM, et al. 2003. Immunocontraception is induced in BALB/c mice inoculated with murine cytomegalovirus expressing mouse zona pellucida 3[J]. Biology of Reproduction, 68(6): 2024-2032.
Luke JM, Vincent JM, Du SX, et al. 2011. Improved antibiotic-free plasmid vector design by incorporation of transient expression enhancers[J]. Gene Therapy, 18(4): 334-343.
Ma JZ, Wang HL, Zheng XX, et al. 2014. CpG/Poly (I∶C) mixed adjuvant priming enhances the immunogenicity of a DNA vaccine against eastern equine encephalitis virus in mice[J]. International Immunopharmacology, 19(1): 74-80.
Ma XL, Kadir Z, Li JY, et al. 2012a. The effects of GM-CSF and IL-5 as molecular adjuvants on immune responses and contraception induced by mZP3 DNA vaccination [J]. American Journal of Reproductive Immunology, 68(6): 476-485.
Ma XL, Li JY, Zhang FC. 2012b. Intranasal co-delivery with the mouse zona pellucida 3 and GM-CSF expressing constructs enhances humoral immune responses and contraception in mice [J]. Scandinavian Journal of Immunology, 76(6): 521-527.
Mackenzie SM, McLaughlin EA, Perkins HD, et al. 2006. Immunocontraceptive effects on female rabbits infected with recombinant myxoma virus expressing rabbit ZP2 or ZP3[J]. Biology of Reproduction, 74(3): 511-521.
Nixon B, Aitken RJ, McLaughlin EA. 2007. New insights into the molecular mechanisms of sperm-egg interaction[J]. Cellular and Molecular Life Sciences, 64(14): 1805-1823.
Peng J, Shi S, Yang Z, et al. 2014. Short noncoding DNA fragments improve the immune potency of electroporation-mediated HBV DNA vaccination[J]. Gene Therapy, 21(7): 703-708.
Robinson HL, Pertmer TM. 2000. DNA vaccines for viral infections: basic studies and applications[J]. Advances in Virus Research, 55: 1-74.
Saiga H,Nieuwenhuizen N, Gengenbacher M, et al. 2015. The recombinant BCG ΔureC∷ hly vaccine targets the AIM2 inflammasome to induce autophagy and inflammation[J]. The Journal of Infectious Diseases, 211(11): 1831-1841.
Sasaki S, Takeshita F, Xin KQ, et al. 2003. Adjuvant formulations and delivery systems for DNA vaccines[J]. Methods, 31(3): 243-254.
Scheerlinck JPY, Karlis J, Tjelle TE, et al. 2004. In vivo electroporation improves immune response to DNA vaccination in sheep[J]. Vaccine, 22(13-14): 1820-1825.
Shahbazi M, Zahedifard F, Saljoughian N, et al. 2015. Immunological comparison of DNA vaccineation using two delivery systems against canine leishmaniasis[J]. Veterinary Parasitology, 212(3-4): 130-139.
Sheng ZY, Gao N, Cui XY, et al. 2016. Electroporation enhances protective immune response of a DNA vaccine against Japanese encephalitis in mice and pigs[J]. Vaccine, 34(47): 5751-5757.
Shrestha A, Srithandan S, Minhas V, et al. 2015. Canine zona pellucida glycoprotein-3: up-scaled production, immunization strategy and its outcome on fertility[J]. Vaccine, 33(1): 133-140.
Shrestha A, Wadhwa N, Gupta SK. 2014. Evaluation of recombinant fusion protein comprising dog zona pellucida glycoprotein-3 and izumo and individual fragments as immunogens for contraception[J]. Vaccine, 32(5): 564-571.
Spargo SC, Hope RM. 2003. Evolution and nomenclature of the zona pellucida gene family[J]. Biology of Reproduction, 68(2): 358-362.
Srivastave N, Santhanam R, Sheela P, et al. 2002. Evaluation of the immunocontraceptive potential of Escherichia coli-expressed recombinant dog ZP2 and ZP3 in a homologous animal model[J]. Reproduction, 123(6): 847-857.
Sun Y, Hu YH, Liu CS, et al. 2010. Construction and analysis of an experimental Streptococcus iniae DNA vaccine[J]. Vaccine, 28(23): 3905-3912.
Talbot P, Shur BD, Myles DG. 2003. Cell adhesion and fertilization: steps in oocyte transport, sperm-zona pellucida interactions, and sperm-egg fusion[J]. Biology of Reproduction, 68(1): 1-9.
Tu YX, Li XP, Kadir Z, et al. 2013. Molecular adjuvant interleukin-33 enhances the antifertility effect of Lagurus lagurus zona pellucida 3 DNA vaccine administered by the mucosal route [J]. Brazilian Journal of Medical and Biological Research, 46(12): 1064-1073.
Ye C, Choi JG, Abraham S, et al. 2015. Targeting DNA vaccines to myeloid cells using a small peptide[J]. European Journal of Immunology, 45(1): 82-88.
CopyRight©2020 Editorial Office of Sichuan Journal of Zoology