Marquez A S, Vicentini A P, Ono M A, Watanabe M A E, de Camargo Z P, Itano E N
Biological and Health Center, UNOPAR and Microbiology Postgraduation Program, UEL, Londrina, Brazil.
J Clin Lab Anal. 2005;19(5):199-204. doi: 10.1002/jcla.20078.
Yeast forms of Paracoccidioides brasiliensis produce polydispersed high molecular mass (h-MM) antigens. We investigated the antibodies to an h-MM antigen from P. brasiliensis by immunoblotting and ELISA in sera from paracoccidioidomycosis (PCM) patients. IgG from the sera of chronic PCM patients was able to recognize the h-MM antigen at a higher frequency in the cell-free antigen (CFA) (8/13) than in the somatic antigen (SA) (2/13), as assessed by immunoblotting. The CFA was fractionated by Sephadex G-200 chromatography, and fraction 17 (F17) with the h-MM antigen of approximately 366 kDa was used in ELISA to analyze specific levels of IgG and IgE. Patients with the chronic form showed significantly higher levels of IgG (P<0.05) but not IgE (P>0.05) to F17 by ELISA, compared to patients with the acute form or to healthy donors. In conclusion, CFA is better than SA as a source of the P. brasiliensis h-MM antigen. This study reveals a new characteristic to differentiate between the acute and chronic forms of PCM, by demonstrating a higher level of seric IgG to h-MM antigen in chronic compared to acute PCM patients.
巴西副球孢子菌的酵母形式可产生多分散的高分子量(h-MM)抗原。我们通过免疫印迹法和酶联免疫吸附测定(ELISA),对来自副球孢子菌病(PCM)患者血清中针对巴西副球孢子菌h-MM抗原的抗体进行了研究。通过免疫印迹法评估,慢性PCM患者血清中的IgG能够识别无细胞抗原(CFA)中的h-MM抗原的频率更高(8/13),高于体细胞抗原(SA)中的频率(2/13)。CFA通过葡聚糖凝胶G-200柱层析进行分级分离,含有约366 kDa h-MM抗原的第17级分(F17)用于ELISA,以分析IgG和IgE的特异性水平。与急性形式的患者或健康供体相比,通过ELISA检测,慢性形式的患者对F17的IgG水平显著更高(P<0.05),但IgE水平无显著差异(P>0.05)。总之,作为巴西副球孢子菌h-MM抗原的来源,CFA优于SA。本研究通过证明慢性PCM患者血清中针对h-MM抗原的IgG水平高于急性PCM患者,揭示了区分PCM急性和慢性形式的一个新特征。