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犬钩虫感染性幼虫和成虫的一种免疫显性16 kDa表面相关抗原——Ac-SAA-1。

Ac-SAA-1, an immunodominant 16 kDa surface-associated antigen of infective larvae and adults of Ancylostoma caninum.

作者信息

Zhan Bin, Wang Yan, Liu Yueyuan, Williamson Angela, Loukas Alex, Hawdon John M, Xue Hae-Chou, Xiao Shu-Hua, Hotez Peter J

机构信息

Department of Microbiology and Tropical Medicine, The George Washington University and Sabin Vaccine Institute, 2300 Eye Street NW, Washington, DC 20037, USA.

出版信息

Int J Parasitol. 2004 Aug;34(9):1037-45. doi: 10.1016/j.ijpara.2004.04.015.

Abstract

A cDNA encoding a surface-associated antigen was cloned from an Ancylostoma caninum infective larvae (L(3)) cDNA library by immunoscreening with pooled human immune sera. The sera were obtained from individuals living in an Ancylostoma duodenale hookworm-endemic region of China, who had light intensity infections and high antibody titers against A. caninum L(3). Ancylostoma caninum surface-associated antigen-1 is encoded by an 843 bp mRNA with a predicted open reading frame of 162 amino acids. Recombinant Ancylostoma caninum surface-associated antigen-1 was expressed in Escherichia coli and used to prepare a specific antiserum. A Western blot with anti-Ancylostoma caninum surface-associated antigen-1 specific antiserum showed that native Ancylostoma caninum surface-associated antigen-1 protein is expressed by both L(3) and adult hookworms; RT-PCR confirmed that the mRNA is transcribed in both stages. In adult hookworms, the protein localised to the basal layer of the cuticle and hypodermis of adult worms. Serological analysis determined that recombinant Ancylostoma caninum surface-associated antigen-1 protein is recognised by 61% of human sera from a Necator americanus hookworm endemic area in China, indicating the antigen is immunodominant. Anti-Ancylostoma caninum surface-associated antigen-1 antiserum partially inhibited (46.7%) invasion of hookworm L(3) into dog skin in vitro. Together these results suggest that Ancylostoma caninum surface-associated antigen-1 offers promise as a protective vaccine antigen.

摘要

通过用人混合免疫血清进行免疫筛选,从犬钩虫感染性幼虫(L(3))cDNA文库中克隆出一个编码表面相关抗原的cDNA。这些血清取自生活在中国十二指肠钩虫流行区、感染程度较轻且对犬钩虫L(3)抗体滴度较高的个体。犬钩虫表面相关抗原-1由一个843 bp的mRNA编码,预测的开放阅读框为162个氨基酸。重组犬钩虫表面相关抗原-1在大肠杆菌中表达,并用于制备特异性抗血清。用抗犬钩虫表面相关抗原-1特异性抗血清进行的蛋白质印迹分析表明,天然犬钩虫表面相关抗原-1蛋白在L(3)和成虫阶段均有表达;逆转录聚合酶链反应(RT-PCR)证实该mRNA在两个阶段均有转录。在成虫中,该蛋白定位于成虫角质层和皮下组织的基底层。血清学分析表明,来自中国美洲板口线虫流行区的61%的人血清能识别重组犬钩虫表面相关抗原-1蛋白,表明该抗原具有免疫优势。抗犬钩虫表面相关抗原-1抗血清在体外能部分抑制(46.7%)钩虫L(3)对犬皮肤的侵袭。这些结果共同表明,犬钩虫表面相关抗原-1有望成为一种保护性疫苗抗原。

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