Morgan Marjorie S, Vyszenski-Moher DiAnn L, Arlian Larry G
Department of Biological Sciences, Wright State University, Dayton, Ohio, USA.
Int Arch Allergy Immunol. 2015;166(4):267-72. doi: 10.1159/000381854. Epub 2015 Apr 29.
The house dust mite, Euroglyphus maynei, occurs in homes worldwide and is an important source of many allergens. Many patients sensitive to Dermatophagoides farinae and D. pteronyssinus are also sensitive to E. maynei. Extracts to detect sensitivity to E. maynei and reagents to detect E. maynei allergens in the environment or in cultures are not readily available. Information for the culture of E. maynei and for the determination of allergen and endotoxin levels in cultures is limited.
We mass cultured E. maynei at 23 and 30°C and determined the population growth profiles from inoculation until cultures could be harvested for the production of extracts. We also developed an ELISA to measure Eur m 1 and Eur m 2 allergens using mouse monoclonal antibodies directed at cross-reacting epitopes of group 1 and group 2 allergens of D. farinae and D. pteronyssinus.
The E. maynei populations grew exponentially at both 23 and 30°C; however, the cultures matured more rapidly at 23°C. The Eur m 1 and Eur m 2 allergen concentrations in culture extracts changed independently as the cultures grew and matured. At both temperatures, the Eur m 1 concentrations increased as the cultures matured, while the Eur m 2 concentrations did not. The endotoxin levels in these cultures were low.
We report here that E. maynei can be cultured at 23 and 30°C. Monoclonal antibodies directed at cross-reacting epitopes on Dermatophagoides allergens can be used to measure the associated E. maynei allergen levels in these cultures.
屋尘螨,梅氏嗜霉螨,在世界各地的家庭中均有出现,是多种过敏原的重要来源。许多对粉尘螨和户尘螨敏感的患者也对梅氏嗜霉螨敏感。用于检测对梅氏嗜霉螨敏感性的提取物以及用于检测环境或培养物中梅氏嗜霉螨过敏原的试剂并不容易获得。关于梅氏嗜霉螨培养以及培养物中过敏原和内毒素水平测定的信息有限。
我们在23℃和30℃下对梅氏嗜霉螨进行大规模培养,并确定了从接种到培养物可收获用于提取物生产期间的种群生长曲线。我们还开发了一种酶联免疫吸附测定法(ELISA),使用针对粉尘螨和户尘螨第1组和第2组过敏原交叉反应表位的小鼠单克隆抗体来测量Eur m 1和Eur m 2过敏原。
梅氏嗜霉螨种群在23℃和30℃下均呈指数增长;然而,培养物在23℃下成熟得更快。培养物提取物中Eur m 1和Eur m 2过敏原浓度随着培养物的生长和成熟而独立变化。在两个温度下,随着培养物成熟,Eur m 1浓度增加,而Eur m 2浓度未增加。这些培养物中的内毒素水平较低。
我们在此报告,梅氏嗜霉螨可在23℃和30℃下培养。针对粉尘螨过敏原交叉反应表位的单克隆抗体可用于测量这些培养物中相关的梅氏嗜霉螨过敏原水平。