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[Immunocontraceptive effect of DNA vaccines of Lagurus lagurus zona pellucida 3 fusioned with C-terminal of HSP70].
Xi Bao Yu Fen Zi Mian Yi Xue za Zhi = Chinese Journal of Cellular and Molecular Immunology 2008 April
AIM: 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).
CONCLUSION: 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.
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).
CONCLUSION: 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.
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