Requena J M, Soto M, Guzman F, Maekelt A, Noya O, Patarroyo M E, Alonso C
Centro de Biología Molecular Severo Ochoa (C.S.I.C.-U.A.M.), Madrid, Spain.
Mol Immunol. 1993 Aug;30(12):1115-21. doi: 10.1016/0161-5890(93)90158-8.
In the present paper we describe the analysis of the immunological recognition by sera of healthy individuals and chagasic patients of the Trypanosoma cruzi heat shock 70 kDa protein. By a Falcon Assay Screening Test, using as antigen an ATP-agarose purified T. cruzi hsp70, it has been found that the sera of infected patients as well as of that of healthy individuals show reactivity against the hsp70 protein but that the reactivity of the sera of patients is in general significantly higher than that of healthy individuals. The analysis of the reactivity of the chagasic sera against a collection of peptides covering 92% of the protein has shown that more than 50% of the peptides gave a positive response but only against a few peptides did we observe high reactivity in a wide spectrum of sera. Only four peptides (numbers 9, 12, 14 and 47) were recognized by all sera tested with high reactivity values. The sera of healthy individuals also showed reactivity against a large percentage of peptides but with lower values. It was observed that particular peptides showing high reactivity against the sera of healthy donors also show high reactivity against patients' sera. However, the general pattern of reactivity against the peptides is different in chagasic and healthy sera. The immunodominant peptides map in the highly conserved as well as in the less conserved part of the hsp70 molecule. The 1/3 C-terminal, being the least conserved part of the molecule, seems to be the least immunogenic. Mapping of the epitopes led to the identification of particular immunogenic motifs within individual peptides.
在本论文中,我们描述了对健康个体和恰加斯病患者血清对克氏锥虫热休克70 kDa蛋白的免疫识别分析。通过福尔肯检测筛选试验,使用ATP-琼脂糖纯化的克氏锥虫hsp70作为抗原,发现感染患者的血清以及健康个体的血清均显示出对hsp70蛋白的反应性,但患者血清的反应性总体上显著高于健康个体。对恰加斯病血清针对覆盖该蛋白92%的一系列肽段的反应性分析表明,超过50%的肽段呈阳性反应,但仅在少数肽段上,我们在广泛的血清中观察到高反应性。所有测试血清均以高反应性值识别出仅四个肽段(编号9、12、14和47)。健康个体的血清对很大比例的肽段也显示出反应性,但反应值较低。观察到对健康供体血清显示高反应性的特定肽段对患者血清也显示出高反应性。然而,恰加斯病血清和健康血清对肽段的总体反应模式不同。免疫显性肽段定位于hsp70分子高度保守以及较不保守的部分。分子中最不保守的1/3 C末端似乎免疫原性最低。表位定位导致在各个肽段中鉴定出特定的免疫原基序。