Immunogenicity Study of Plasmid DNA Encoding Mouse Cysteine-Rich Secretory Protein-1 (mCRISP1) as a Contraceptive Vaccine
Article first published online: 19 MAR 2012
© 2012 John Wiley & Sons A/S
American Journal of Reproductive Immunology
Volume 68, Issue 1, pages 47–55, July 2012
How to Cite
- Issue published online: 5 JUN 2012
- Article first published online: 19 MAR 2012
- Manuscript Accepted: 3 FEB 2012
- Manuscript Received: 25 NOV 2011
- National Population and Family Planning Commission of Hubei Province Foundation Project in China. Grant Number: JS-2010002
- self-research programme for Doctoral Candidates (including Mphil-PhD) of Wuhan Unversity. Grant Number: 20103020201000187
- DNA vaccine;
To examine the immunocontraceptive properties of the plasmid pcDNA-mCRISP1 and compare them to the corresponding recombinant mCRISP1 (r-mCRISP1).
Method of study
RT-PCR and indirect immunofluorescence were performed to observe the mCRISP1 protein expression in COS-7 cells. Three groups of mice received three injections of r-mCRISP1, pcDNA-mCRISP1 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-mCRISP1 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.
COS-7 cells transfected with pcDNA-mCRISP1 present the expression of mCRISP1. Both r-mCRISP1 and pcDNA-mCRISP1 raised an immune response against r-mCRISP1 protein and native CRISP1 in mouse sperm. The titres of anti-mCRISP1 antibodies from DNA immunized mice were significantly lower than that of r-mCRISP1 immunized mice, but it lasted relatively longer. Male and female pcDNA-mCRISP1 injected animals presented a statistically significant reduction in their fertility with no signs of immunopathologic effects.
These studies demonstrated the feasibility of generating an immune response to mCRISP1 protein by DNA vaccine and pcDNA-mCRISP1 plasmid causing significant anti-fertility potential.