中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
24期
1905-1908
,共4页
黄慧聪%姚丽丽%周子博%郑丽娜%潘长旺
黃慧聰%姚麗麗%週子博%鄭麗娜%潘長旺
황혜총%요려려%주자박%정려나%반장왕
管圆线虫,广州%表位%电脉,凝胶,双向%免疫印迹
管圓線蟲,廣州%錶位%電脈,凝膠,雙嚮%免疫印跡
관원선충,엄주%표위%전맥,응효,쌍향%면역인적
Angiostrongylus cantonensis%Epitopes%Electrophoresis,gel,two-dimensional%Western blot
目的 寻找广州管圆线虫Ⅴ期幼虫可溶性蛋白中具免疫反应性的抗原.方法 采用双向电泳结合免疫印迹技术,并将得到的蛋白点进行质谱分析,获得具免疫原性的肽段信息,合成肽段免疫小鼠并进一步观察其体外免疫结合性能.结果 Ⅴ期雌雄虫可见的印迹反应点数目分别为8个和4个,此12个点相对应的凝胶点pH值集中于5～7,相对分子质量范围为24 000～40 000.质谱鉴定分析肌动蛋白和半乳糖凝集素为Ⅴ期雌雄虫的共同抗原.进一步对半乳糖凝集素进行抗原表位预测,获得16肽(NH2-KNGDIALHFNPRFDEK)表位,Western印迹验证16肽的小鼠免疫血清能识别广州管圆线虫Ⅴ期幼虫32 000的抗原表位.结论 半乳糖凝集素和肌动蛋白为Ⅴ期雌雄虫具有免疫反应性的共同抗原,半乳糖凝集素16肽抗原表位可作为靶标用于免疫诊断技术的开发和免疫调节机制的研究.
目的 尋找廣州管圓線蟲Ⅴ期幼蟲可溶性蛋白中具免疫反應性的抗原.方法 採用雙嚮電泳結閤免疫印跡技術,併將得到的蛋白點進行質譜分析,穫得具免疫原性的肽段信息,閤成肽段免疫小鼠併進一步觀察其體外免疫結閤性能.結果 Ⅴ期雌雄蟲可見的印跡反應點數目分彆為8箇和4箇,此12箇點相對應的凝膠點pH值集中于5～7,相對分子質量範圍為24 000～40 000.質譜鑒定分析肌動蛋白和半乳糖凝集素為Ⅴ期雌雄蟲的共同抗原.進一步對半乳糖凝集素進行抗原錶位預測,穫得16肽(NH2-KNGDIALHFNPRFDEK)錶位,Western印跡驗證16肽的小鼠免疫血清能識彆廣州管圓線蟲Ⅴ期幼蟲32 000的抗原錶位.結論 半乳糖凝集素和肌動蛋白為Ⅴ期雌雄蟲具有免疫反應性的共同抗原,半乳糖凝集素16肽抗原錶位可作為靶標用于免疫診斷技術的開髮和免疫調節機製的研究.
목적 심조엄주관원선충Ⅴ기유충가용성단백중구면역반응성적항원.방법 채용쌍향전영결합면역인적기술,병장득도적단백점진행질보분석,획득구면역원성적태단신식,합성태단면역소서병진일보관찰기체외면역결합성능.결과 Ⅴ기자웅충가견적인적반응점수목분별위8개화4개,차12개점상대응적응효점pH치집중우5～7,상대분자질량범위위24 000～40 000.질보감정분석기동단백화반유당응집소위Ⅴ기자웅충적공동항원.진일보대반유당응집소진행항원표위예측,획득16태(NH2-KNGDIALHFNPRFDEK)표위,Western인적험증16태적소서면역혈청능식별엄주관원선충Ⅴ기유충32 000적항원표위.결론 반유당응집소화기동단백위Ⅴ기자웅충구유면역반응성적공동항원,반유당응집소16태항원표위가작위파표용우면역진단기술적개발화면역조절궤제적연구.
Objective To identify the specific antigens of the fifth stage larvae (larvae Ⅴ) of Angiostrongylus cantonensis in soluble proteins.Methods The techniques of two-dimensional electrophoresis (2-DE) and immunoblot were employed to analyze the soluble components of larvae Ⅴ antigens.The resulting protein spots were identified by mass spectrometry and the specific antigen and immunogenic peptide segment profiles determined.Larvae Ⅴ was identified by immunoblot of peptide immune murine sera.Results Immunoblot of 2-DE showed that there were 8 specific antigen spots in females and 4 in males of larvae Ⅴ.The 12 protein spots had a pH range of 5-7 and a molecular weight of 24-40 kD.Among 12 spot,actin-1 and galectin-1 were identified successfully.And a peptide(NH2-KNGDIALHFNPRFDEK) was predicted as epitope and synthesized.The binding capacity of murine immune sera could identify the 32 000 of larvae Ⅴ.Conclusions Galectin and actin are common antigens of female and male larvae Ⅴ of A.cantonensis.And a peptide from galectin-1 may become a potential target for immunologic diagnosis and immunological mediator.
