- 作者: Ching-Yun Chang a, Hong Chou a, Ming F. Tam b, Ren-BinTang c, Hsiu-Yu Lai a, Horng-Der Shen a
- 作者服務機構: Departments of a Medical Research and Education and c Pediatrics, Veterans General Hospital-Taipei , b Institute of Molecular Biology , Academia Sinica , Taipei , Taiwan , ROC
- 中文摘要: --
- 英文摘要: Rhodotorula mucilaginosa (also known as R. rubra) isamong the most commonly found yeast strains in ourenvironment. However, allergens from R. mucilaginosahave not yet been characterized at the molecular level.The purpose of this study was to characterize the enolaseallergen from R. mucilaginosa and examine the aller-genic/antigenic cross-reactivity among fungal enolases.The full-length cDNA encoding the R. mucilaginosa eno-lase was isolated through the reverse transcriptase-poly-merase chain reaction in conjunction with the 5'-end and3'-end rapid amplification cDNA end reactions. The cor-responding natural enolase from R. mucilaginosa wasidentified using two-dimensional gel electrophoresisand N-terminal amino acid sequence analysis. The re-suits showed that the enolase from R. mucilaginosa is aprotein of 439 residues and is encoded by a cDNA of1, 497 by. It shares high sequence identity with enolaseallergens from Candida albicans (85%), Saccharomycescerevisiae (76%), Penicillium citrinum (76%). Aspergillusfumigatus (76%), Cladosporium herbarum (76.5%), andAltemaria alternata (74%). A 47-kD component in theR. mucilaginosa extracts was found to react with IgE orrabbit anti-enolase antiserum and has an N-terminalamino acid sequence identical to that deduced from theisolated enolase cDNA. Sera from three (21%) of 14 aller-gic patients sensitized to R. mucilaginosa showed lgEbinding to this 47-kD R. mucilaginosa component andthe His-tagged recombinant enolase. A rabbit antiserumagainst the P. citrinum enolase and a monoclonal anti-body (MoAb; Afueno 8) against the A. fumigatus enolasereacted with all 5 fungal enolases tested. However, anMoAb (E2a) generated by using the Saccharomyces eno-lase as antigen could only recognize the immunizingenolase. In addition, heterogeneity in immunoblot pro-files of IgE antibodies in serum samples from 9 allergicpatients against 5 different fungal enolases tested wasalso observed. The presence of IgE cross-reactivityamong enolase allergens from R. mucilaginosa, C. albi-cansand P. citrinumwas detected by immunoblot inhibi-tion. In conclusion, a new and cross-reactive enolaseallergen from R. mucilaginosa (Rho m 1) was identified.Although enolases are highly conserved allergensamong different fungal species, most of the allergicpatients examined in this study differed in their IgE reac-tivity to the 5 different fungal enolases tested. The resultsobtained will be of value in understanding the role ofenolase allergen in clinical mould allergy.
- 中文關鍵字: --
- 英文關鍵字: Rhodotorula mucilaginosa, Enolase, cDNA, Monoclonal antibody, IgE cross-reactivity