HSP70C融合草原兔尾鼠卵透明带3对小鼠抗生育的研究  

Immunocontraceptive effect of DNA vaccines of Lagurus lagurus zona pellucida 3 fusioned with C-terminal of HSP70

作　　者：张钰[1] 李轶杰[1] 马正海[1] 陈璇[1] 潘晓燕[2] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046 [2]新疆畜牧科学院,新疆乌鲁木齐830000

出　　处：《细胞与分子免疫学杂志》2008年第4期354-357,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30360062);新疆教育厅创新群体资助项目(XJEDU2004G02)

摘　　要：目的:探讨分枝杆菌热休克蛋白(HSP70)对草原兔尾鼠卵透明带3(LZP3)免疫不育效果的增强作用。方法:将HSP70全长和C端分别与LZP3融合构建pcD-L-HSP70和pcD-L-HSP70C重组质粒,同时与具有免疫不育功能的pcD-L和pcD-ACLC一起分别免疫NIH小鼠,分别检测重组质粒在机体真核细胞中的表达情况、T细胞增殖及体液免疫水平、抗生育效果、卵巢的病理变化和免疫荧光定位。结果:LZP3构建的重组质粒均能在小鼠肝脏中表达;免疫后能刺激机体T细胞增殖,尤其是pcD-ACLC和pcD-L-HSP70C免疫组(P<0·01);同时激发机体产生特异性抗体(P<0·05);除pcD-L-HSP70免疫组外其他3种重组质粒均具有抗生育效果(P<0·05),且pcD-ACLC和pcD-L-HSP70C极明显地降低了小鼠平均窝仔数(P<0·01)且未引发机体产生卵巢炎;直接免疫荧光结果表明在绿色激发光下小鼠卵母细胞的卵透明带均能发出绿色荧光。结论:与pcD-L-HSP70免疫相比,pcD-L-HSP70C能明显地降低小鼠平均窝仔数,这表明分枝杆菌热休克蛋白HSP70C端对增强LZP3免疫不育效果具有明显的基因佐剂功效。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 hydrnamic 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 lymphccytes in all groups were proliferated by stimulating the recombinant LZP3 with no disruption of follicular in ovaties, 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 pcDACLC and pcD-L-HSP70C （ P 〈 0. 01 ）. CONCLUSION： LZP3 fused by the C-terminal of mycobacterium tuberculosis heat shock protein HSP70 had a good immunocontraceptiveeffect 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.

关 键 词：草原兔尾鼠卵透明带3 免疫不育 HSP70 融合佐剂 

分 类 号：Q78[生物学—分子生物学]