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[澳洲群钩端螺旋体属特异性单克隆抗体的制备、鉴定及其抗原检测研究]

[Study on the production, identification of serogroup-specific monoclonal antibodies against leptospires of Australis serogroup and detection of its antigen].

作者信息

Liang J, Dai B

出版信息

Hua Xi Yi Ke Da Xue Xue Bao. 1990 Sep;21(3):234-8.

PMID:1709413
Abstract

Spleen cells of BALB/c mice immunized with whole Leptospira interrogans serogroup Australis serovar australis strain 620 were fused with myeloma cells line SP2/0. Specificities of four McAbs determined by MAT. 2E1 McAb (IgG3) reacted with 11 serovars, of the Australis serogroup, but did not react with 22 representative serovars of L. interrogans in 20 serogroups, L. biflexa strain patoc I and Leptonema illini strain 3055. 2E1 McAb showed serogroup specificity for Australis by agglutination and the other 3 McAbs showed partial serogroup specificity. We compared the outer envelope (OE) protein profiles of serovar australis strain 620 with those of two pathogenic L. interrogans serovar lai strain 601 and serovar hebdomadis strain 156 by SDS-PAGE. 63kd protein profile was only found in the OE of strain 620, and the quantity of 42kd protein of strain 620 was greater than that of strain 601 and 156. The immunoblotting revealed that 2E1 McAb reacted with a 34kd band in the OE preparation of serovar australis strain 620, but did not react with that of other two L. interrogans. 2E1 McAb also did not react with OE of non-pathogenic leptospires. It was suggested that 34kd protein might contain the antigenic determinants which were shared by leptospires of Australis serogroup.

摘要

用澳洲钩端螺旋体血清群澳洲型菌株620免疫的BALB/c小鼠的脾细胞与骨髓瘤细胞系SP2/0进行融合。通过显微镜凝集试验(MAT)确定了四种单克隆抗体(McAb)的特异性。2E1 McAb(IgG3)与澳洲钩端螺旋体血清群的11个血清型发生反应,但不与问号钩端螺旋体20个血清群中的22个代表性血清型、双曲钩端螺旋体菌株帕托c I及伊利诺伊钩端螺旋体菌株3055发生反应。2E1 McAb通过凝集反应显示出对澳洲钩端螺旋体血清群的血清型特异性,其他3种McAb显示出部分血清群特异性。我们通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)比较了澳洲型菌株620与两种致病性问号钩端螺旋体赖型菌株601和七日热型菌株156的外膜(OE)蛋白图谱。63kd蛋白图谱仅在菌株620的外膜中发现,菌株620的42kd蛋白量大于菌株601和156。免疫印迹显示,2E1 McAb与澳洲型菌株620外膜制剂中的一条34kd条带发生反应,但不与其他两种问号钩端螺旋体的外膜制剂发生反应。2E1 McAb也不与非致病性钩端螺旋体的外膜发生反应。提示34kd蛋白可能含有澳洲钩端螺旋体血清群钩端螺旋体共有的抗原决定簇。

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