钩端螺旋体澳洲群群特异性McAb的建立、鉴定及群特异性抗原检测

Study on the Production, Identification of Serogroup-Specific Monoclonal Antibodies against Leptospires of Australis Serogroup and Detection of its Antigen

作者成功地获得4株分泌抗澳洲群钩瑞螺旋体的单克隆抗体(McAb)杂交瘤。其中2E_1株能与所选澳洲群的11个型的钩体都发生凝集反应,而不与澳洲群外的20群22个型的钩体、非致病性Patot Ⅰ株钩体和伊利尼细螺旋体3055株发生凝集反应。由此表明,2E_1 McAb具有澳洲群钩体的群特异性。用SDS-PAGE、免疫印迹法对澳洲型620株、赖型601株、七日热型156株、Patoc Ⅰ株及3055株外膜蛋白中McAb识别的抗原成分进行检测,结果2E_1 McAb能特异地识别澳洲型620株34kd外膜蛋白区带。其余3株McAb具有澳洲群钩体部分群特异性。

Spleen cells of BALB/c mice immunizedwith whole Leptospira interrogans serogroupAustralis serovar australis strain 620 werefused with myeloma cells line SP_(2/0).Specificities ot four McAbs determinedby MAT. 2E_1 McAb (IgG_3) reacted with11 serovars. or the Australis serogroup,but did not react with 22 representativeserovars of L. interrogans in 20 serogroups,L. biflexa strain patoc I and Leptonemaillini strain 3055. 2E_1 McAb showed sero-group speciticity for Australis by aggluti-nation and the other 3 McAbs showedpartial serogroup specificity. We compared the puter envelope (OE)protein profiles ot serovar australis strain630 with those of two pathogenic L.inter-rogans serovar lai strain 601 and serovarhebdomadis strain 156 by SDS-PAGE. 63kdprotein profile was only found in the OEof strain 620, and the quantity of 42kdprotein of strain 62o was greater thanthat of strain 601 and 156. The immunoblotting revealed that 2E_1McAb reacted with a 34kd band in the OEpreparation of serovar australis strain 620.but did not react with that of other twoL. interrogans. 2E_1 McAb also did rot reactwith OE of non--pathogenic leptospires.It was suggested that 34kd protein mightcontain the antigenic determinants whichwere shared by leptospires of Australisserogroup.