Umezawa E S, Shikanai-Yasuda M A, Stolf A M
Instituto de Medicina, Tropical de São Paulo, Faculdade de Medicina-Universidade de São Paulo, Brazil.
J Clin Lab Anal. 1996;10(6):407-13. doi: 10.1002/(SICI)1098-2825(1996)10:6<407::AID-JCLA16>3.0.CO;2-0.
This report describes differences in humoral immune response of acute and chronic phases of human Chagas disease. The reactivities of IgG, IgM, and IgA anti-Trypanosoma cruzi antibodies in serum samples from both groups of patients were compared by enzyme-linked immunosorbent assay (ELISA) employing either one of four antigenic fractions: mouse laminin (LAM), which reacts through Gal alpha 1-3Gal epitopes expressed on trypomastigote surface: whole intact trypomastigotes (TCT); trypomastigotes excreted/secreted antigens (TESA); and epimastigote alkaline extract (EAE). The selection of T. cruzi antigen preparations was based on their relative content of surface and internal antigens found in trypomastigote forms. The proportion of IgG reactive to carbohydrate epitopes was assessed through the decay of IgG reactivity from acute and chronic sera after m-periodate oxidation of solid-phase bound antigens. Trypomastigote and TESA antigens recognized by IgG from acute and chronic sera were also compared by immunoblotting. ELISA and immunoblotting data showed that: (1) the proportion of IgG directed to trypomastigote surface antigens was higher in acute than in chronic sera, whereas the opposite was found for internal antigens, (2) acute sera contained a higher percentage of IgG reactive to trypomastigote carbohydrate epitopes than chronic sera, and (3) anti-T. cruzi IgA was found exclusively in acute sera and led to 100% positivity when LAM, TCT, and TESA were employed as antigens. IgA ELISA with these antigens and IgG immunoblotting pattern with TESA could be useful as serological markers for the acute phase of human Chagas disease.
本报告描述了人类恰加斯病急性期和慢性期体液免疫反应的差异。通过酶联免疫吸附测定(ELISA),使用四种抗原成分之一,比较了两组患者血清样本中IgG、IgM和IgA抗克氏锥虫抗体的反应性:小鼠层粘连蛋白(LAM),其通过锥鞭毛体表面表达的Galα1-3Gal表位发生反应;完整的全锥鞭毛体(TCT);锥鞭毛体排泄/分泌抗原(TESA);以及前鞭毛体碱性提取物(EAE)。克氏锥虫抗原制剂的选择基于其在锥鞭毛体形式中发现的表面和内部抗原的相对含量。通过对固相结合抗原进行高碘酸盐氧化后,急性和慢性血清中IgG反应性的衰减,评估与碳水化合物表位反应的IgG比例。还通过免疫印迹比较了急性和慢性血清中IgG识别的锥鞭毛体和TESA抗原。ELISA和免疫印迹数据表明:(1)针对锥鞭毛体表面抗原的IgG比例在急性血清中高于慢性血清,而内部抗原的情况则相反;(2)急性血清中与锥鞭毛体碳水化合物表位反应的IgG百分比高于慢性血清;(3)抗克氏锥虫IgA仅在急性血清中发现,当使用LAM、TCT和TESA作为抗原时,阳性率达100%。使用这些抗原的IgA ELISA和使用TESA的IgG免疫印迹模式可作为人类恰加斯病急性期的血清学标志物。