Kobayashi Y, Tamai T, Oyama T, Hasegawa H, Sada E, Kusaba T, Hamaji M
Microbiol Immunol. 1984;28(3):359-70. doi: 10.1111/j.1348-0421.1984.tb00687.x.
Monoclonal antibodies against etiological agents of Weil's disease were produced by cell fusion technology. Twenty hybridomas were produced through the fusion of P3X63Ag8 .653 cells with spleen cells from BALB/c mice immunized against Leptospira interrogans serovar icterohaemorrhagiae RGA strain and serovar copenhageni Shiromizu and M20 strains. Reactivities of the antibodies produced by the hybridomas were determined by the microscopic agglutination test. Among the five hybridoma antibodies to the RGA strain, two reacted specifically to serovar icterohaemorrhagiae, two reacted to serovar icterohaemorrhagiae at a high titer and serovar copenhageni at a low titer, and one reacted to serovars icterohaemorrhagiae, copenhageni, pyrogens, and canicola. Of the ten hybridoma antibodies to the Shiromizu strain, one reacted specifically to serovar copenhageni, seven reacted to both serovars copenhageni and icterohaemorrhagiae at almost the same titer, and two exhibited intermediate properties. Of the five hybridoma antibodies to the M20 strain, three reacted to both serovars copenhageni and icterohaemorrhagiae at almost the same titer, one reacted to serovar copenhageni at a low titer and serovar icterohaemorrhagiae at a high titer, and one reacted to serovars copenhageni, icterohaemorrhagiae, and pyrogens. The results revealed that each serovar has its own antigen(s) and their common antigens. In addition, 20 strains of leptospires were recently isolated and tested with three monoclonal antibodies characterized by different reactivities. Twenty strains were clearly identified by their antibodies, i.e., 16 strains were identified as serovar icterohaemorrhagiae and three strains were identified as serovar copenhageni. The remaining strain, which was not agglutinated by three antibodies, was identified as serovar autumnalis by an agglutination test with immune rabbit sera.
通过细胞融合技术制备了针对魏氏病病原体的单克隆抗体。将P3X63Ag8.653细胞与经黄疸出血型钩端螺旋体RGA菌株、哥本哈根型钩端螺旋体Shiromizu和M20菌株免疫的BALB/c小鼠的脾细胞融合,产生了20个杂交瘤。通过显微镜凝集试验测定杂交瘤产生的抗体的反应性。在针对RGA菌株的5种杂交瘤抗体中,2种特异性地与黄疸出血型血清型反应,2种以高滴度与黄疸出血型血清型反应、以低滴度与哥本哈根型血清型反应,1种与黄疸出血型、哥本哈根型、致热型和犬型血清型反应。在针对Shiromizu菌株的10种杂交瘤抗体中,1种特异性地与哥本哈根型血清型反应,7种以几乎相同的滴度与哥本哈根型和黄疸出血型血清型反应,2种表现出中间特性。在针对M20菌株的5种杂交瘤抗体中,3种以几乎相同的滴度与哥本哈根型和黄疸出血型血清型反应,1种以低滴度与哥本哈根型血清型反应、以高滴度与黄疸出血型血清型反应,1种与哥本哈根型、黄疸出血型和致热型血清型反应。结果表明,每种血清型都有其自身的抗原和它们的共同抗原。此外,最近分离了20株钩端螺旋体并用三种具有不同反应性特征的单克隆抗体进行检测。通过它们的抗体清楚地鉴定出了20株,即16株被鉴定为黄疸出血型血清型,3株被鉴定为哥本哈根型血清型。其余一株未被三种抗体凝集,通过与免疫兔血清的凝集试验被鉴定为秋季型血清型。