Bush R K, Sanchez H
William S. Middleton Memorial Veterans Administration Hospital, Madison, Wisconsin, USA.
Ann Allergy Asthma Immunol. 1997 Mar;78(3):287-92. doi: 10.1016/S1081-1206(10)63183-3.
Allergic reactions to fungal allergens, including Alternaria, are common clinical problems. Reagents used for diagnosis and therapy of fungal allergy are complex and variable mixtures. Standardized reagents are difficult to achieve due to batch to batch variability in these biologic products. Although several Alternaria allergens have been isolated, their production by current methods is laborious. The introduction of molecular biology into allergy research has led to the molecular cloning of several allergens which may culminate in improved methods of treatment.
This report describes the development of techniques that will lead to the molecular cloning of Alternaria allergens.
We isolated the poly (A)(+)-messenger RNA from Alternaria, produced proteins in vitro and probed them for binding to murine monoclonal antibodies directed against Alternaria. In addition, we examined the ability of the translated proteins to bind IgE from the sera of Alternaria-sensitive individuals.
We synthesized at least 20 proteins ranging in molecular weight from 2 to 90 kD using in vitro techniques. The translated proteins were detected by both murine monoclonal and human IgE antibodies.
Alternaria allergens can be synthesized in vitro by molecular biology techniques. Such techniques will be used in the development of cDNA libraries for the production of Alternaria allergens by recombinant DNA methods.
对包括链格孢属在内的真菌过敏原的过敏反应是常见的临床问题。用于真菌过敏诊断和治疗的试剂是复杂且可变的混合物。由于这些生物制品批次间的变异性，难以实现标准化试剂。尽管已经分离出几种链格孢属过敏原，但通过现有方法生产它们很费力。将分子生物学引入过敏研究已导致几种过敏原的分子克隆，这可能最终带来改进的治疗方法。
本报告描述了将导致链格孢属过敏原分子克隆的技术的发展。
我们从链格孢属中分离出聚腺苷酸（+）信使核糖核酸，在体外产生蛋白质，并检测它们与针对链格孢属的鼠单克隆抗体的结合情况。此外，我们研究了翻译后的蛋白质与链格孢属敏感个体血清中IgE结合的能力。
我们使用体外技术合成了至少20种分子量从2到90千道尔顿不等的蛋白质。翻译后的蛋白质可被鼠单克隆抗体和人IgE抗体检测到。
可通过分子生物学技术在体外合成链格孢属过敏原。此类技术将用于构建cDNA文库，以便通过重组DNA方法生产链格孢属过敏原。
