Lustigman S, Brotman B, Huima T, Prince A M
Lindsley F. Kimball Research Institute, New York Blood Center, New York 10021.
Mol Biochem Parasitol. 1991 Mar;45(1):65-75. doi: 10.1016/0166-6851(91)90028-5.
The isolation and characterization of a recombinant cDNA clone (OV7) expressing an antigen present in Onchocerca volvulus infective larvae and adult stages is described. Using chimpanzee antiserum generated against irradiated infective larvae, we isolated a cDNA clone from a lambda gt11 cDNA expression library derived from adult O. volvulus mRNA. The open reading frame encodes 131 amino acids corresponding to a 15.2-kDa protein. Affinity purified antibodies which bound specifically to OV7 fusion polypeptide recognized a single antigen with an apparent molecular weight of 17,000 in extracts of L3, L4 and adult worms. Immunoelectron microscopy established that the antigen encoded by this clone is present in the hypodermis and the basal layer of the cuticle of L3 and female adult worm, and in the egg shell around developing microfilariae. Since the OV7 fusion polypeptide is onchocerca-specific and is recognized specifically by sera from onchocerciasis patients, and sera from non-patent but infected chimpanzees, and not by sera from patients with other filarial parasites, it may have potential as an antigenic component in a test for detection of non-patent and patent infections of O. volvulus. The OV7 amino acid sequence contains residues that have a probable homology with the cysteine proteinase inhibitor superfamily.
本文描述了一个重组cDNA克隆(OV7)的分离和特性,该克隆表达盘尾丝虫感染性幼虫和成虫阶段存在的一种抗原。利用针对经辐照的感染性幼虫产生的黑猩猩抗血清,我们从一个源自成年盘尾丝虫mRNA的λgt11 cDNA表达文库中分离出一个cDNA克隆。开放阅读框编码131个氨基酸,对应一个15.2 kDa的蛋白质。与OV7融合多肽特异性结合的亲和纯化抗体在L3、L4和成虫提取物中识别出一种表观分子量为17,000的单一抗原。免疫电子显微镜证实,该克隆编码的抗原存在于L3和成年雌虫的皮下组织和角质层的基底层,以及发育中的微丝蚴周围的卵壳中。由于OV7融合多肽具有盘尾丝虫特异性,能被盘尾丝虫病患者血清以及未出现症状但已感染的黑猩猩血清特异性识别,而不能被其他丝虫寄生虫患者血清识别,因此它可能有潜力作为一种抗原成分用于检测盘尾丝虫的未出现症状和已出现症状的感染。OV7氨基酸序列包含与半胱氨酸蛋白酶抑制剂超家族可能具有同源性的残基。