Monoclonal antibodies to parasite antigens found in the serum of Dirofilaria immitis-infected dogs.
Monoclonal antibodies to parasite antigens found in the serum of Dirofilaria immitis-infected dogs.
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在感染恶丝虫的狗的血清中发现了针对寄生虫抗原的单克隆抗体。
作者：
G. Weil;M. Malane;K. G. Powers;L. S. Blair
我们最近报道，寄生虫抗原检测犬心丝虫感染的犬血清中的对流免疫电泳（CIE）。通过用部分纯化的抗原制剂免疫小鼠，将脾细胞与SP-2骨髓瘤细胞融合，并通过ELISA和CIE抑制筛选细胞培养上清液中的抗体，获得产生对这些抗原特异性的单克隆抗体的杂交瘤细胞系。鉴定了两种主要循环寄生虫抗原共有的两个表位的特异性抗体。免疫过氧化物酶研究表明，单克隆抗体识别的抗原表位在D. immitis，但女性子宫和卵子特别强烈的标记。建立了一种基于单克隆抗体的ELISA检测犬血清中寄生虫抗原的方法。46只感染犬血清中有45只检测到寄生虫抗原，但未感染犬血清和感染隐匿性双瓣线虫犬血清中均未检测到寄生虫抗原。血清抗原含量与从感染犬中回收的雌蠕虫数量显著相关（r = 0.82，p <0.001）。感染后6个月首次检测到抗原血症，感染后9至21个月期间抗原水平保持相当稳定。寄生虫抗原检测与这种单克隆抗体为基础的ELISA似乎是上级的灵敏度，特异性和感染强度的关系，以前描述的诊断方法犬恶丝虫病。
We recently reported that parasite antigens are detectable in the serum of Dirofilaria immitis-infected dogs by counterimmunoelectrophoresis (CIE). Hybridoma cell lines that produce monoclonal antibodies specific for these antigens were obtained by immunizing mice with a partially purified antigen preparation, fusing spleen cells with SP-2 myeloma cells, and screening cell culture supernatants for antibody by ELISA and CIE inhibition. Antibodies specific for two epitopes shared by the two major circulating parasite antigens were identified. Immunoperoxidase studies showed that the epitopes recognized by the monoclonals were widely distributed in D. immitis, but the female uterus and eggs were particularly strongly labeled. A monoclonal antibody-based ELISA was developed to measure parasite antigens in dog sera. Parasite antigens were detected in 45 of 46 sera from infected dogs but were absent in sera from uninfected dogs and sera from dogs infected with Dipetalonema reconditum. Serum antigen content was significantly correlated with the number of female worms recovered from infected dogs (r = 0.82, p less than 0.001). Antigenemia was first detected 6 mo after infection, and antigen levels remained fairly stable between 9 and 21 mo after infection. Parasite antigen detection with this monoclonal antibody-based ELISA appears to be superior to previously described diagnostic methods for canine dirofilariasis in terms of sensitivity, specificity, and relation to infection intensity.