Madan Taruna, Priyadarsiny Priyanka, Vaid Mudit, Kamal Neel, Shah Ashok, Haq Wahajul, Katti Seturam Bandacharya, Sarma P Usha
Molecular Biochemistry and Diagnostics, Institute of Genomics and Integrative Biology, Mall Rd., Delhi, India.
Clin Diagn Lab Immunol. 2004 May;11(3):552-8. doi: 10.1128/CDLI.11.3.552-558.2004.
Allergic bronchopulmonary aspergillosis (ABPA) is an immunologically complex allergic disorder caused by the fungal pathogen Aspergillus fumigatus. Elevated levels of total immunoglobulin E (IgE), specific IgE, and IgG antibodies in sera are important immunodiagnostic criteria for ABPA. International reference standards or standardized immunodiagnostic assays are not available due to a lack of well-defined diagnostic antigens. The present study was carried out to identify and evaluate the immunodiagnostic relevance of synthetic epitopic peptides of Asp f 1, a major allergen, antigen, or cytotoxin of A. fumigatus. Five overlapping peptides were synthesized from the N terminus of Asp f 1, one of the potential immunodominant regions predicted by algorithmic programs. The 11-amino-acid synthetic peptide (P1) significantly inhibited both IgG binding (89.10% +/- 4.45%) and IgE binding (77.32% +/- 3.38%) of the standardized diagnostic antigen (SDA) (a well-defined pool of diagnostically relevant allergens and antigens of A. fumigatus). With a panel of sera of ABPA patients, allergic patients with skin test negativity to A. fumigatus, and healthy individuals, P1 showed a higher diagnostic efficiency than SDA (specific IgG, 100%; specific IgE, 98.3%). The diagnostic efficiency of P1 could be attributed to the presence of homologous epitopes in various immunodominant allergens or antigens of A. fumigatus. The ability of P1 to induce histamine release from sensitized mast cells and a Th2 type of cytokine profile in peripheral blood mononuclear cells of ABPA patients suggests its potential for use in intradermal testing. P1 could be further explored for development of a standardized, specific, and sensitive immunodiagnostic test for aspergillosis.
变应性支气管肺曲霉病(ABPA)是一种由真菌病原体烟曲霉引起的免疫复杂的变应性疾病。血清中总免疫球蛋白E(IgE)、特异性IgE和IgG抗体水平升高是ABPA重要的免疫诊断标准。由于缺乏明确的诊断抗原,尚无国际参考标准或标准化免疫诊断检测方法。本研究旨在鉴定和评估烟曲霉主要变应原、抗原或细胞毒素Asp f 1的合成表位肽的免疫诊断相关性。从Asp f 1的N端合成了5个重叠肽,Asp f 1是算法程序预测的潜在免疫优势区域之一。11个氨基酸的合成肽(P1)显著抑制标准化诊断抗原(SDA)(一组明确的烟曲霉诊断相关变应原和抗原)的IgG结合(89.10%±4.45%)和IgE结合(77.32%±3.38%)。对于一组ABPA患者、烟曲霉皮肤试验阴性的变应性患者和健康个体的血清,P1显示出比SDA更高的诊断效率(特异性IgG为100%;特异性IgE为98.3%)。P1的诊断效率可归因于烟曲霉各种免疫优势变应原或抗原中存在同源表位。P1诱导致敏肥大细胞释放组胺以及ABPA患者外周血单个核细胞中Th2型细胞因子谱的能力表明其在皮内试验中的应用潜力。P1可进一步用于开发标准化、特异性和敏感的曲霉病免疫诊断检测方法。