Immunoglobulin E-binding and skin test reactivity to hydrophobin HCh-1 from Cladosporium herbarum, the first allergenic cell wall component of fungi

BACKGROUND: For many years, fungal spores have been recognized as potential causes of respiratory allergies. All fungal allergens cloned so far represent either secreted or cytoplasmatic proteins, but nothing is known about the involvement of fungal surface proteins in allergic diseases. METHODS: A phage surface displayed cDNA-library from the mould Cladosporium herbarum was constructed and phage displaying IgE-binding proteins were selectively enriched with immobilized serum IgE from C. herbarum-sensitized individuals. Inserts encoding putative allergens were sequenced, subcloned and used to produce recombinant proteins. Allergenicity of the proteins was evaluated by IgE binding in Western blots, enzyme-linked immunosorbent assay (ELISA) and skin prick test in a total of 84 patients sensitized to either C. herbarum or Aspergillus fumigatus and three healthy controls. RESULTS: After four rounds of affinity selection, the cDNA-library was enriched for clones displaying IgE-binding molecules. Sequencing of inserts showed that one clone contained an open reading frame predicting a protein of 105 amino acids and a calculated molecular weight of 10.5 kDa showing the classical signature of members of the hydrophobin family. The recombinant protein, termed HCh-1, was able to bind IgE from six patients sensitized to fungi in vitro. Two of those patients were also included in a skin prick test survey and showed strong type I skin reactions to HCh-1, demonstrating the allergenic nature of C. herbarum hydrophobin and indicating a prevalence of sensitization in the range of 8-9%. In contrast, the hydrophobin HYP1 from Aspergillus fumigatus was not recognized by the sera of the same patients and controls investigated with HCh-1. CONCLUSION: C. herbarum hydrophobin represents the first component of the cell wall of fungi demonstrated to act as a rare but clinically relevant allergen in vitro and in vivo.     

Fungi and allergic lower respiratory tract diseases

Asthma is a common disorder that in 2009 afflicted 8.2% of adults and children, 24.6 million persons, in the United States. In patients with moderate and severe persistent asthma, there is significantly increased morbidity, use of health care support, and health care costs. Epidemiologic studies in the United States and Europe have associated mold sensitivity, particularly to Alternaria alternata and Cladosporium herbarum, with the development, persistence, and severity of asthma. In addition, sensitivity to Aspergillus fumigatus has been associated with severe persistent asthma in adults. Allergic bronchopulmonary aspergillosis (ABPA) is caused by A fumigatus and is characterized by exacerbations of asthma, recurrent transient chest radiographic infiltrates, coughing up thick mucus plugs, peripheral and pulmonary eosinophilia, and increased total serum IgE and fungus-specific IgE levels, especially during exacerbation. The airways appear to be chronically or intermittently colonized by A fumigatus in patients with ABPA. ABPA is the most common form of allergic bronchopulmonary mycosis (ABPM); other fungi, including Candida, Penicillium, and Curvularia species, are implicated. The characteristics of ABPM include severe asthma, eosinophilia, markedly increased total IgE and specific IgE levels, bronchiectasis, and mold colonization of the airways. The term severe asthma associated with fungal sensitization (SAFS) has been coined to illustrate the high rate of fungal sensitivity in patients with persistent severe asthma and improvement with antifungal treatment. The immunopathology of ABPA, ABPM, and SAFS is incompletely understood. Genetic risks identified in patients with ABPA include HLA association and certain T(H)2-prominent and cystic fibrosis variants, but these have not been studied in patients with ABPM and SAFS. Oral corticosteroid and antifungal therapies appear to be partially successful in patients with ABPA. However, the role of antifungal and immunomodulating therapies in patients with ABPA, ABPM, and SAFS requires additional larger studies.     

Sensitization to Alternaria and Cladosporium by the age of 4 years

Of 1218 children born on the Isle of Wight in 1989/90, and followed for atopy at age 4 years, 981 were skin-prick tested witha battery of allergens. Of these 61 (6%) reacted positively to Alternaria alternata and Cladosporium herbarum (47 to Alternaria, 21 to Cladosporium and seven to both). Twenty-four (39%) were asymptomatic (latent atopy) of which 12 had a single positive reaction either to Alternaria or Cladosporium. Asthma was the most common disease in children sensitized to moulds, Alternaria sensitization correlated positively with clinical diagnosis of asthma (P < 0.01), eczema (P <0.001) and rhinitis (P <0.05), Likewise, Cladosporium sensitivity correlated with a diagnoses of asthma, eczema and rhinitis (all P <0.05). Age of the house correlated with reported damp and lack of central heating (both P <0.001), but not with sensitization to moulds. An association between the presence of damp or age of the house and mould allergy was confounded by 21 children moving house in the first 4 years. Exposure to pets, passive tobacco smoking and season of birth had no bearing on mould sensitivity. At 4 years of age Alternaria and Cladosporium were the third most common causes of sensitization, i.e. after house dust mite and grass pollen.     

卡氏枝孢霉与阿耶罗枝孢瓶霉的随机扩增DNA多态性研究

研究卡氏枝孢霉与阿耶罗枝孢瓶霉的ＤＮＡ多态性，了解两菌在ＤＮＡ型别与形态学及菌种来源地域的相互关系。采用溴苯提取法提取ＤＮＡ，以随机扩增ＤＮＡ多态性方法对来自五个国家的２１株分离株（１９株卡氏枝孢霉，２株阿耶罗枝孢瓶霉）进行研究。结果：①１０个随机引物筛选出３个具有较好ＤＮＡ扩增的引物。②１９株卡氏枝孢霉的ＤＮＡ带型具有较大的遗传相似性，阿耶罗枝孢瓶霉的ＤＮＡ带型与卡氏枝孢霉略有不同，但与中国株具有较大相似性（８５％的遗传相似性）。③卡氏枝孢霉的遗传相似性与菌株的来源地域关系密切，同一国家的菌株具有相似的ＤＮＡ带型，位于树状图的同一树组中。结论：随机扩增ＤＮＡ多态性研究发现卡氏枝孢霉具有一定的种内差异，其ＤＮＡ带型与菌株的来源地域关系密切，阿耶罗枝孢瓶霉的ＤＮＡ带型与某些株卡氏枝孢霉更相似。本研究方法简便、快速，可用于流行病学及基因学特征的研究     

Histologic und Elektronenmikroskopie der Tinea nigra/Light and Electron Microscopic Investigation of Tinea Nigra

Zusammenfassung: Bei einem häufig beruflich in Südamerika beschäftigten 50jährigen Mann wurde eine Tinea nigra plantaris klinisch und histologisch diagnostiziert. Der Herd hatte sich seit 3 Jahren langsam peripherwärts ausgebreitet. Klinisch ähnelte er einem beginnenden akrolentiginösen Melanom. Subjektive Symptome bestanden nicht. Der Patient konnte den schmutzig braunen Fleck mit einem Bimsstein nahezu vollständig abreiben. Durch die histologische Untersuchung wurde die klinische Verdachtsdiagnose der Tinea nigra gesichert. Unter lokaler antimykotischer Behandlung mit Clotrimazol heilte die Tinea nigra ab.
Summary: Tinea nigra plantaris was diagnosed in a 50 year old man who had frequently been in South America. The asymptomatic lesion had slowly enlarged for 3 years. It was clinically similar to early acral lentiginous melanoma. The patient had been able to remove the pigmentation nearly completely by abrasive measures. Histology proved the suspected clinical diagnosis of tinea nigra. Topical clotrimazole treatment cleared the lesion.     

Diagnosis and immunotherapy of mould allergy

The IgE-response was evaluated by skin prick test, bronchial provocation test and RAST in a 1-year placebo-controlled double-blind immunotherapy study. Eleven adult asthmatics were treated with a Cladosporium allergen preparation and 11 comparable patients received histamine placebo. The bronchial sensitivity (PC20) decrease greater than 0.5 log step in 8/11 (73%) Cladosporium-treated versus 3/11 (27%) in the placebo group. Corresponding figures for skin prick test sensitivity was 10/11 (91%) and 1/11 (9%) respectively. Circulating IgE showed a temporary boost in the Cladosporium group and then values approaching the pretreatment value. Only minimal and insignificant changes were found in the placebo-treated patients. Changes in IgE-reactivity were not related to allergen dose, clinical efficacy or to the occurrence of side effects. Some interrelation between changes in skin prick test, bronchial provocation test and RAST was found indicating a differentiated effect of immunotherapy on various IgE compartments. In spite of the pathogenetic role of IgE in allergic diseases, changes in IgE-reactivity do not seem directly involved in the mechanisms underlying the clinical efficacy of immunotherapy but might be of importance in a complex interaction with other immunological parameters.     

Successful treatment of chromoblastomycosis with itraconazole

Summary. An unusual severe case of chromoblastomycosis due to Cladosporium carrionii unresponsive to 5-FC and some azoles is reported. With oral itraconazole at a dosage of 100 mg d^-1 for 15 months (total dose 45.5 g) the patient had a complete clinical and mycological recovery without any side-effects.
Zusammenfassung. Es wird von einem ungewöhnlich schweren Fall einer Chromoblastomykose aufgrund einer Infektion mit Cladosporium carrionii berichtet, die nicht auf 5-FC und einige Azol-Antimykotika ansprach. Daraufhin wurde 15 Monate lang 100 mg/d Itraconazol gegeben. Die Gesamtdosis betrug 45,5 g. Beim Patienten zeigten sich keine Nebenwirkungen, und er heilte klinisch und mykologisch vollständig aus.     

小鼠皮肤卡氏支孢霉感染动物模型的构建

目的 构建卡氏支孢霉感染小鼠模型.方法 ICR小鼠分为4组,A组为健康小鼠,足垫皮下接种1 × 108孢子数/mL卡氏支孢霉悬液组;B组为免疫抑制小鼠,足垫皮下接种1 × 108孢子数/mL卡氏支孢霉悬液组;C组为免疫抑制小鼠足垫皮下接种1 × 106孢子数/mL卡氏支孢霉悬液组;D组为对照组,健康小鼠足垫皮下接种生理氯化钠溶液.接种后第7、30、60天时测量各组小鼠足垫厚度,处死并进行足垫病理及真菌检查.结果 A、B、c组小鼠接种处均出现肿胀、发黑、溃疡、结痂,感染发病率均为100%.A组足垫厚度在接种后第7天、30天分别为(2.85±0.47)mm、(2.40±0.45)mm,后者较前者显著下降(P<0.05);第60天为(1.64±0.13)mm,较第30天显著下降(P<0.05).B组在接种后第7天、30天分别为(1.80±0.21)mm、(2.19±0.27)mm,后者较前者显著增厚(P<0.05);第60天为(1.86±0.22)mm,较第30天显著下降(P<0.05).C组在接种后第7天、30天分别为(1.51±0.11)mm、(1.98±0.06)mm,后者较前者显著增厚(P<0.05),第60天为(1.82±0.09)mm,较第30天显著下降(P<0.05). D组则无显著改变(P>0.05).实验组基本病理改变为坏死、脓肿、慢性肉芽肿形成.HE、PAS染色可见硬壳小体,皮损处脓液直接镜检可见硬壳小体,培养为卡氏支孢霉生长.对照组未感染卡氏支孢霉.结论 卡氏支孢霉鼠足垫皮下接种免疫抑制和健康小鼠均可成功建立皮下着色真菌病模型.