[Immunocontraceptive effect of DNA vaccines of Lagurus lagurus zona pellucida 3 fusioned with C-terminal of HSP70].
OBJECTIVE
To investigate the immunocontraceptive effect of Lagurus lagurus zona pellucida 3 gene (LZP3) fused by Mycobacterium tuberculosis HSP70 and C-terminal of HSP70.


METHODS
Two immunocontractive recombinant plasmids pcD-L-HSP70 and pcD-L-HSP70C were constructed using LZP3. Meanwhile, pcD-L and pcD-ACLC which were effective immunocontraceptive vaccines were studied as control.


RESULTS
After the LZP3 genes were expressed in the livers of mice with the introduction of hydronamic transfection instead of traditional HeLa cell transfection by RT-PCR, NIH mice were immunized with ZP3 DNA vaccines. The results of ELISA showed that all of the recombinants in mice elicited specific anti-ZP3 antibodies which were combined with the ZP3 of oocytes from the immunized mice in immunofluorescence assay. MTS assay showed that the lymphocytes in all groups were proliferated by stimulating the recombinant LZP3 with no disruption of follicular in ovaries, especially those in groups of pcD-ACLC and pcD-L-HSP70C (P<0.01). Antifertility experiments showed that three groups (besides pcD-L-HSP70) enhanced the sterile effects (P<0.05), especially those in the groups of pcD-ACLC and pcD-L-HSP70C (P<0.01).


CONCLUSIONS
LZP3 fused by the C-terminal of mycobacterium tuberculosis heat shock protein HSP70 had a good immunocontraceptive effect while LZP3 fused by Mycobacterium tuberculosis heat shock protein HSP70 had a poor effect on birth control. So the C-terminal of Mycobacterium tuberculosis heat shock protein HSP70 might be a better candidate to deliver the adjuvant function in LZP3 DNA immunocontraceptive vaccination than the complete HSP70 molecule.
Zhang
Yu
Y
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China. zhangyu_xj@yahoo.com.cn
Li
Yi-jie
YJ
Ma
Zheng-hai
ZH
Chen
Xuan
X
Pan
Xiao-yan
XY
Zhang
Fu-chun
FC
D000276:Adjuvants, Immunologic / Q000235:genetics / Q000378:metabolism; D000818:Animals; D003411:Arvicolinae / Q000378:metabolism; D004527:Egg Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005260:Female; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011956:Receptors, Cell Surface / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000072216:Zona Pellucida Glycoproteins
D000276:Adjuvants, Immunologic; D004527:Egg Proteins; D018840:HSP70 Heat-Shock Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D000072216:Zona Pellucida Glycoproteins; C000606521:Zp3 protein, mouse