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1、南京农业大学学报 2002,25(4):67～71JournalofNanjingAgriculturalUniversity猪链球菌2型溶菌酶释放蛋白的黏附作用1,213曾巧英,陆承平(11南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏南京210095;21甘肃农业大学动物医学院,甘肃兰州730070)摘要:为研究猪链球菌2型(SS2)的毒力因子溶菌酶释放蛋白(MRP)的黏附作用,进行了如下试验:11用黏附计数法+-+比较菌株HA9801(MRP)和SH006444(MRP)的黏附动力学,两菌株均能黏附于HEp2细胞,MRP株的最大黏附菌+数显著
2、高于MRP株(P<0105)。21菌体置56℃1h,或用胰酶或半乳糖预处理,细菌黏附力完全丧失;分别用高碘酸、溶菌酶、秋水仙素和DNaseⅠ处理HEp2细胞,前者可完全阻断HA9801对它的黏附,后三者使黏附菌数显著下降。+31用纯化的MRP处理HEp2细胞,或用抗MRP兔血清或MRP的单克隆抗体处理菌体,MRP株的黏附菌数显著降低-+(P<0105),但不能被完全抑制;用抗MRP株全菌兔血清处理菌体,也能降低MRP株的黏附菌数。41半乳糖或56℃预处理MRP,使其完全丧失对HA9801的黏附抑制功能,但胰酶、过氧化氢和巯基乙醇的预处理没有明显的影响。结论:
3、MRP是SS2的一种黏附素,半乳糖可阻断其黏附作用。关键词:猪链球菌2型;黏附特性;黏附素;溶菌酶释放蛋白中图分类号:S941142 文献标识码:A 文章编号:10002030(2002)04006705RoleasadhesinofmuramidasereleasedproteinofStreptococcussuistype21,213ZENGQiao2ying,LUCheng2ping(11KeyLaboratoryofAnimalDiseaseDiagnosticandImmunology,MinistryofAgriculture,Nanj
4、ingAgricUniv,Nanjing210095,China;21DeptofAnimalMedicine,GansuAgricUniv,Lanzhou730070,China)Abstract:Theadhesionofmuramidasereleasedprotein(MRP),avirulencefactorofStreptococcussuistype2(SS2)weretested.The+-adhesivekineticsofSS2isolateHA9801(MRP)andSH006444(MRP)werecomparedthroughadh
5、esivecounting.Bothstrainscould+-adheretoHEp22cells,buttheadhesivelevelsofMRPstrainweresignificantlyhigherthanthatofMRPstrain(P<0105).Thead2hesiveactivitiesofHA9801wereabolishedbypretreatmentofbacteriumcellwithtrypsinorgalactose,orat56℃1h,andbypretreat2mentofHEp22cellswithperiodicac
6、id.AfterpretreatmentofHEp22cellswithcolchicine,lysozymeorDNaseⅠ,theadhesivelevels+couldbereduceddramatically.TheadhesiveactivitiesofMRPstrainwereobviouslyreducedbutnotblockedbypretreatmentofbacterum-cellswithantiserumormonoclonalantibodiesagainstMRP,oragainstMRPstrain,aswellasbypre
7、treatmentofHEp22cellswithpuri2fiedMRP.MRPwasdestroyedcompletelyitsinhibitiveperformanceagainstadhesionofHA9801toHEp22cellsafterpretreatmentat56℃orwithgalactose,otherthanwithtrypsin,mercaptoethanolandhydrogenperoxide.Conclusion:MRPisanadhesinofSS2andgalactosecanblockitsadhesion.Keyw
8、ords:Streptococcussuistype
