Schmechel Detlef, Simpson Janet P, Beezhold Donald, Lewis Daniel M
National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services, Morgantown, WV 26505, USA.
J Immunol Methods. 2006 Feb 20;309(1-2):150-9. doi: 10.1016/j.jim.2005.12.001. Epub 2006 Jan 3.
Mold contamination and exposure to fungi in indoor environments has been associated with various adverse health effects but little is known about the significance of individual fungal species in the initiation or exacerbation of such effects. Using Stachybotrys chartarum as a model fungus we sought to demonstrate that monoclonal antibodies (mAbs) can provide species-specific diagnostic reagents and also be used to investigate immunological cross-reactivity patterns among fungi. Mice were immunized with S. chartarum spore walls and monoclonal antibodies were screened against 60 fungal species and 24 different isolates of S. chartarum using an indirect ELISA. One species-specific mAb (IgG(1)) reacted only with spore preparations but not mycelium of S. chartarum or propagules of any other fungus. Five cross-reactive mAbs (IgM) documented extensive cross-reactivity among nine related Stachybotrys species and several non-related genera including several species of Cladosporium, Memnoniella, Myrothecium and Trichoderma. We also found that the ELISA reactivity for cross-reactive antigens and different isolates of S. chartarum differed considerably for normalized total amounts of mycelial antigen. We demonstrate that mAbs and immunoassays have the potential to detect S. chartarum species-specifically. The observed reactivity patterns with cross-reactive mAbs suggest that several fungi may share common antigens and that the majority of antigens are expressed by spores and mycelia. The observed cross-reactivity patterns need to be considered for accurate interpretations of environmental and serological analyses.
室内环境中的霉菌污染和真菌暴露与多种不良健康影响相关，但对于单个真菌物种在引发或加剧此类影响中的重要性却知之甚少。我们以炭疽杆菌作为模式真菌，试图证明单克隆抗体（mAb）可提供物种特异性诊断试剂，还可用于研究真菌之间的免疫交叉反应模式。用炭疽杆菌孢子壁免疫小鼠，并使用间接ELISA针对60种真菌物种和24种不同的炭疽杆菌分离株筛选单克隆抗体。一种物种特异性单克隆抗体（IgG(1)）仅与炭疽杆菌的孢子制剂反应，而不与炭疽杆菌的菌丝体或任何其他真菌的繁殖体反应。五种交叉反应性单克隆抗体（IgM）记录了九种相关炭疽杆菌物种以及几个非相关属（包括几种枝孢属、单端孢霉属、漆斑菌属和木霉属物种）之间广泛的交叉反应。我们还发现，对于菌丝体抗原的标准化总量，交叉反应性抗原和不同炭疽杆菌分离株的ELISA反应性差异很大。我们证明单克隆抗体和免疫测定法有潜力特异性地检测炭疽杆菌。观察到的与交叉反应性单克隆抗体的反应模式表明，几种真菌可能共享共同抗原，并且大多数抗原由孢子和菌丝体表达。为了准确解释环境和血清学分析，需要考虑观察到的交叉反应模式。
