Matsumoto Tokiko K, Cotrim Paulo C, da Silveira José Franco, Stolf Anna Maria S, Umezawa Eufrosina S
Instituto Adolfo Lutz-São Paulo, Brazil.
Diagn Microbiol Infect Dis. 2002 Mar;42(3):187-92. doi: 10.1016/s0732-8893(01)00348-0.
Trypomastigote forms of Trypanosoma cruzi excrete-secrete several molecules, which are immunodominant during the human infection. This complex antigenic mixture termed TESA (Trypomastigote Excreted-Secreted Antigens) presents a 150-160 kDa band that shows excellent specificity and sensitivity in Chagas' disease diagnosis by immunoblotting. Here we describe the isolation and the antigenic characterization of a recombinant peptide (TESA-1) containing a 10 kDa T. cruzi peptide that belongs to the 150-160 kDa TESA fraction. The clone was isolated by screening a T. cruzi genomic expression library with chagasic antibodies reactive to the 150-160 kDa band of TESA immunoblots. After expression, the recombinant peptide TESA-1 was purified and used to immunize rabbits. Anti-TESA-1 immunesera specifically recognized the 150-160 kDa fraction of TESA-blots from eight different T. cruzi strains. The TESA-1 peptide reacted with 82.2% of chagasic patient sera by immunoblotting, showing that it harbors most of the antigenic epitopes that account for the high reactivity of the 150-160 kDa band of TESA.
克氏锥虫的锥鞭毛体形式分泌多种分子，这些分子在人类感染过程中具有免疫显性。这种称为TESA（锥鞭毛体分泌抗原）的复杂抗原混合物呈现出一条150 - 160 kDa的条带，在通过免疫印迹法诊断恰加斯病时显示出优异的特异性和敏感性。在此，我们描述了一种重组肽（TESA - 1）的分离及其抗原特性，该重组肽包含一个10 kDa的克氏锥虫肽，属于150 - 160 kDa的TESA组分。通过用对TESA免疫印迹的150 - 160 kDa条带具有反应性的恰加斯病抗体筛选克氏锥虫基因组表达文库来分离该克隆。表达后，纯化重组肽TESA - 1并用于免疫兔子。抗TESA - 1免疫血清特异性识别来自八个不同克氏锥虫菌株的TESA印迹的150 - 160 kDa组分。通过免疫印迹法，TESA - 1肽与82.2%的恰加斯病患者血清发生反应，表明它含有大部分抗原表位，这些表位导致了TESA的150 - 160 kDa条带的高反应性。
