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Immunogenicity Study of Plasmid DNA Encoding Mouse Cysteine‐Rich Secretory Protein‐1 ( mCRISP 1) as a Contraceptive Vaccine
Author(s) -
Luo Jin,
Yang Jing,
Cheng Yan,
Li Wei,
Yin TaiLang,
Xu WangMing,
Zou YuJie
Publication year - 2012
Publication title -
american journal of reproductive immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.071
H-Index - 97
eISSN - 1600-0897
pISSN - 1046-7408
DOI - 10.1111/j.1600-0897.2012.01117.x
Subject(s) - immunogenicity , biology , dna vaccination , immune system , western blot , antibody , recombinant dna , immunofluorescence , plasmid , sperm , antiserum , microbiology and biotechnology , andrology , virology , immunology , immunization , dna , gene , medicine , biochemistry , botany , genetics
Problem To examine the immunocontraceptive properties of the plasmid pcDNA ‐ mCRISP 1 and compare them to the corresponding recombinant mCRISP 1 ( r‐mCRISP 1). Method of study RT‐PCR and indirect immunofluorescence were performed to observe the mCRISP 1 protein expression in COS ‐7 cells. Three groups of mice received three injections of r‐mCRISP 1, pcDNA ‐ mCRISP 1 or pcDNA vector, respectively. ELISA and Western blot were used to examine the immune responses and immunoreactivity of antisera. Sperm‐egg penetration assay was performed to examine the effect of anti‐ mCRISP 1 antibodies in vitro fertilization of mouse oocytes. Fertility and mean litter size were analysed by natural mating. Histological analysis was carried out to look for potential immunopathologic effects of the antibodies. Results COS ‐7 cells transfected with pcDNA ‐ mCRISP 1 present the expression of mCRISP1 . Both r‐mCRISP1 and pcDNA ‐ mCRISP 1 raised an immune response against r‐mCRISP 1 protein and native CRISP 1 in mouse sperm. The titres of anti‐ mCRISP 1 antibodies from DNA immunized mice were significantly lower than that of r‐mCRISP 1 immunized mice, but it lasted relatively longer. Male and female pcDNA ‐ mCRISP 1 injected animals presented a statistically significant reduction in their fertility with no signs of immunopathologic effects. Conclusion These studies demonstrated the feasibility of generating an immune response to mCRISP 1 protein by DNA vaccine and pcDNA ‐ mCRISP 1 plasmid causing significant anti‐fertility potential.