Construction of sperm-specific lactate dehydrogenase DNA vaccine and experimental study of its immunocontraceptive effect on mice.
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.
Chen
Yong
Y
Research Laboratory of Medical Genetics, PLA Institute of Technology for Birth Control, Fetal Health and Child Growth, Fuzhou General Hospital of Nanjing Military Region, Fuzhou 350025, China. fhlan005@163.com
Zhang
Duo
D
Xin
Na
N
Xiong
YongZhong
Y
Chen
Ping
P
Li
Bo
B
Tu
XiangDong
X
Lan
FengHua
F
D000818:Animals; D003269:Contraception, Immunologic / Q000379:methods*; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D007114:Immunization; D007527:Isoenzymes / Q000235:genetics / Q000276:immunology; D007770:L-Lactate Dehydrogenase / Q000235:genetics / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012726:Sexual Behavior, Animal; D013073:Sperm Agglutination; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000166:cytology / Q000201:enzymology*; D019444:Vaccines, DNA
D007527:Isoenzymes; D019444:Vaccines, DNA; D007770:L-Lactate Dehydrogenase; C409673:lactate dehydrogenase C4