国际医学寄生虫病杂志
國際醫學寄生蟲病雜誌
국제의학기생충병잡지
INTERNATIONAL JOURNAL OF MEDICAL PARASITIC DISEASES
2011年
3期
135-139
,共5页
程娜%许学年%张鸿满%周岩%谭裕光%包意芳%张陆娟%徐斌%江河%黎学铭%冯正
程娜%許學年%張鴻滿%週巖%譚裕光%包意芳%張陸娟%徐斌%江河%黎學銘%馮正
정나%허학년%장홍만%주암%담유광%포의방%장륙연%서빈%강하%려학명%풍정
华支睾吸虫%富甘氨酸2a抗原%抗原性%组织定位%分泌蛋白
華支睪吸蟲%富甘氨痠2a抗原%抗原性%組織定位%分泌蛋白
화지고흡충%부감안산2a항원%항원성%조직정위%분비단백
Clonorchis sinensis%Glycine rich antigen 2a%Antigenicity%Histolocalization%Secretory protein
目的 对华支睾吸虫富甘氨酸2a(glycine rich antigen 2a,GRA2a)类抗原Cs4重组蛋白(rCs4)进行免疫学特性分析,并对GKA2a类抗原进行组织定位.方法 使用蛋白质印迹(Western印迹)分析rCs4与华支睾吸虫病患者混合血清的免疫反应性,并分析GRA2a类抗原的分泌性.应用ELISA法检测rCs4免疫小鼠血清中抗rCs4特异性IgG水平,并动态检测华支睾吸虫感染兔0～44周的血清中抗rCs4特异性IgG水平.通过免疫荧光实验(immunofluorescence assay,IFA)对华支睾吸虫GRA2a类抗原进行组织定位.结果 纯化的rCsd可被华支睾吸虫病患者混合血清识别.rCs4单抗(mAb Cs4-31)与华支睾吸虫排泄分泌抗原和可溶性抗原在相对分子质量(Mr)26 000与55 000之间分别有13、15个反应条带,均呈阶梯状分布.ELISA检测rCs4免疫小鼠抗rCs4和ESA的IgG效价均达1:64 000,感染兔的抗rCs4特异性抗体平均水平自感染第4周开始上升,于第6周达到高峰,此后逐步递减,至第20周已呈较低水平.IFA检测发现GRA2a类抗原定位于华支睾吸虫成虫的表皮.结论 Cs4蛋白是华支睾吸虫在感染早期的分泌型蛋白,具较好的抗原性.GRA2a类抗原定位于华支睾吸虫成虫表皮.
目的 對華支睪吸蟲富甘氨痠2a(glycine rich antigen 2a,GRA2a)類抗原Cs4重組蛋白(rCs4)進行免疫學特性分析,併對GKA2a類抗原進行組織定位.方法 使用蛋白質印跡(Western印跡)分析rCs4與華支睪吸蟲病患者混閤血清的免疫反應性,併分析GRA2a類抗原的分泌性.應用ELISA法檢測rCs4免疫小鼠血清中抗rCs4特異性IgG水平,併動態檢測華支睪吸蟲感染兔0～44週的血清中抗rCs4特異性IgG水平.通過免疫熒光實驗(immunofluorescence assay,IFA)對華支睪吸蟲GRA2a類抗原進行組織定位.結果 純化的rCsd可被華支睪吸蟲病患者混閤血清識彆.rCs4單抗(mAb Cs4-31)與華支睪吸蟲排洩分泌抗原和可溶性抗原在相對分子質量(Mr)26 000與55 000之間分彆有13、15箇反應條帶,均呈階梯狀分佈.ELISA檢測rCs4免疫小鼠抗rCs4和ESA的IgG效價均達1:64 000,感染兔的抗rCs4特異性抗體平均水平自感染第4週開始上升,于第6週達到高峰,此後逐步遞減,至第20週已呈較低水平.IFA檢測髮現GRA2a類抗原定位于華支睪吸蟲成蟲的錶皮.結論 Cs4蛋白是華支睪吸蟲在感染早期的分泌型蛋白,具較好的抗原性.GRA2a類抗原定位于華支睪吸蟲成蟲錶皮.
목적 대화지고흡충부감안산2a(glycine rich antigen 2a,GRA2a)류항원Cs4중조단백(rCs4)진행면역학특성분석,병대GKA2a류항원진행조직정위.방법 사용단백질인적(Western인적)분석rCs4여화지고흡충병환자혼합혈청적면역반응성,병분석GRA2a류항원적분비성.응용ELISA법검측rCs4면역소서혈청중항rCs4특이성IgG수평,병동태검측화지고흡충감염토0～44주적혈청중항rCs4특이성IgG수평.통과면역형광실험(immunofluorescence assay,IFA)대화지고흡충GRA2a류항원진행조직정위.결과 순화적rCsd가피화지고흡충병환자혼합혈청식별.rCs4단항(mAb Cs4-31)여화지고흡충배설분비항원화가용성항원재상대분자질량(Mr)26 000여55 000지간분별유13、15개반응조대,균정계제상분포.ELISA검측rCs4면역소서항rCs4화ESA적IgG효개균체1:64 000,감염토적항rCs4특이성항체평균수평자감염제4주개시상승,우제6주체도고봉,차후축보체감,지제20주이정교저수평.IFA검측발현GRA2a류항원정위우화지고흡충성충적표피.결론 Cs4단백시화지고흡충재감염조기적분비형단백,구교호적항원성.GRA2a류항원정위우화지고흡충성충표피.
Objective To study immunological characteristics of Cs4 recombinant protein(rCs4)of Clonorchis sinensis,one of the glycine rich antigen 2a(GRA2a)gene family member,and the histolocalization of GRA2a antigens. Methods The immunological reactivity of Clonorcihs sinensis Cs4 protein to pooled sera from clonorchiasis patients was investigated by using purified rCs4 in Western blotting.The secretory property of GRA2a antigens was also analyzed by Western blotting.The level of specific antibodies against rCs4 in the sera from mice immunized with rCs4,and the dynamic change of antibodies against rCs4 in the sera of C. sinensisinfected New Zealand rabbits(O-44 w post infection)were examined by ELISA.Immunofluorescence assay (IFA)was performed using the anti-rCs4 monoclonal antibody Cs4-31(mAb Cs4-31)to determine the localization of GRA2a antigens in C. sinensis adult worm.Results The purified rCs4 was recognized by pooled sera from clonorchiasis patients.13 and 15 bands appeared at the sites between M,26 000 and 55 000 after mAb Cs4-31 reacting with excretory-secretory antigen(ESA)and soluble adult worm antigen(AWA),respectively.The anti-rCs4 and anti-ESA serum antibody titers in mice immunized with rCs4 were 1:64 000.In C. sinensis infected rabbits,antibody level began to elevate at 4 w after infection,peaked at 6 w,and declined rapidly to a lower level at 20 w.IFA revealed that GRA2a antigens were localized in the tegument of C.sinensis adult worm.Conclusion The Cs4 protein was a secretory antigen appeared in the early stage of infection,and exhibited high antigenieity.GRA2a antigens were localized in tIle tegument of C. sinensis adult worm.
