hsp70c融合草原兔尾鼠卵透明带3对小鼠抗生育的研究

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1、HSP70C融合草原兔尾鼠卵透明带3对小鼠抗生育的研究作者：张钰,李轶杰,马正海,陈璇,潘晓燕,张富春【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

2、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

3、端对增强LZP3免疫不育效果具有明显的基因佐剂功效。【关键词】草原兔尾鼠卵透明带3免疫不育HSP70融合佐剂15　　［Abstract］AIM:ToinvestigatetheimmunocontraceptiveeffectofLaguruslaguruszonapellucida3gene(LZP3)fusedbyMycobacteriumtuberculosisHSP70andC-terminalofHSP70.METHODS:TwoimmunocontractiverecombinantplasmidspcD-L-HSP70andpcD-L-HSP

4、70CwereconstructedusingLZP3.Meanwhile,pcD-LandpcD-ACLCwhichwereeffectiveimmunocontraceptivevaccineswerestudiedascontrol.RESULTS:AftertheLZP3geneswereexpressedintheliversofmicewiththeintroductionofhydrnamictransfectioninsteadoftraditionalHeLacelltransfectionbyRT-PCR,NIHmicewereimmu

5、nizedwithZP3DNAvaccines.TheresultsofELISAshowedthatalloftherecombinantsinmiceelicitedspecificanti-ZP3antibodieswhichwerecombinedwiththeZP3ofoocytesfromtheimmunizedmiceinimmunofluorescenceassay.MTSassayshowedthatthelymphocytesinallgroupswereproliferatedbystimulatingtherecombinantLZ

6、P3withnodisruptionoffollicularinovaries,especiallythoseingroupsofpcD-ACLCandpcD-L-HSP70C(P<0.01).Antifertilityexperimentsshowedthatthreegroups(besidespcD-L-HSP70)enhancedthesterileeffects(P<0.05),especiallythoseinthegroupsofpcD-ACLCandpcD-L-HSP70C(P<0.01).CONCLUSION:LZP3f

7、usedbythe15C-terminalofmycobacteriumtuberculosisheatshockproteinHSP70hadagoodimmunocontraceptiveeffectwhileLZP3fusedbyMycobacteriumtuberculosisheatshockproteinHSP70hadapooreffectonbirthcontrol.SotheC-terminalofMycobacteriumtuberculosisheatshockproteinHSP70mightbeabettercandidateto

8、delivertheadjuvantfunctioninLZP3D