Valenta R, Breiteneder H, Petternburger K, Breitenbach M, Rumpold H, Kraft D, Scheiner O
Institute of General and Experimental Pathology, University of Vienna, Austria.
J Allergy Clin Immunol. 1991 Mar;87(3):677-82. doi: 10.1016/0091-6749(91)90388-5.
To investigate the relationship of the major allergens of birch (Bet v I), alder (Aln g I), hazel (Cor a I), and hornbeam (Car b I) at the nucleic acid level, a cDNA clone coding for the complete Bet v I protein was used for Northern and Southern blot experiments. RNAs were isolated from pollen of birch (Betula verrucosa), alder (Alnus glutinosa), hazel (Corylus avellana), and hornbeam (Carpinus betulus). Hybridization was performed at different stringencies. At high stringency, comparable binding of the complete Bet v I cDNA probe to pollen RNAs from birch, alder, and hazel could be observed, indicating high homology of the mRNAs coding for these allergens. With the 3' and 5' half fragments of the Bet v I cDNA, both probes bound to transcripts of all four tree pollens, but most strongly to birch RNA. In Southern blots, distinct binding patterns of genomic DNA digests of birch, alder, hazel, and hornbeam were observed. Most bands were observed with birch DNA digests and less with alder, whereas in genomic DNA digests of hornbeam and hazel, only one band was observed. The result of these cross-hybridization experiments indicate a high homology at the nucleic acid level of the four major allergens of trees belonging to the order Fagales. The sequence similarity presented here further corroborates earlier observations of immunologic cross-reactivity at the protein level. Therefore, in the case of the major allergens of the Betulaceae, an extract with only one major allergen, preferentially Bet v I, instead of all four major allergens, should be sufficient for diagnostic and therapeutic purposes.
为了在核酸水平上研究桦树(Bet v I)、桤木(Aln g I)、榛树(Cor a I)和鹅耳枥(Car b I)主要过敏原之间的关系,使用编码完整Bet v I蛋白的cDNA克隆进行Northern和Southern印迹实验。从桦树(疣枝桦)、桤木(欧洲桤木)、榛树(欧洲榛)和鹅耳枥(欧洲鹅耳枥)的花粉中分离RNA。在不同的严谨度下进行杂交。在高严谨度下,可以观察到完整的Bet v I cDNA探针与桦树、桤木和榛树花粉RNA有相当的结合,表明编码这些过敏原的mRNA具有高度同源性。使用Bet v I cDNA的3'和5'半片段,两种探针都与所有四种树木花粉的转录本结合,但与桦树RNA的结合最强。在Southern印迹中,观察到桦树、桤木、榛树和鹅耳枥基因组DNA消化物的不同结合模式。桦树DNA消化物观察到的条带最多,桤木较少,而在鹅耳枥和榛树的基因组DNA消化物中,只观察到一条带。这些交叉杂交实验的结果表明,属于壳斗目树木的四种主要过敏原在核酸水平上具有高度同源性。这里呈现的序列相似性进一步证实了早期在蛋白质水平上免疫交叉反应性的观察结果。因此,对于桦木科的主要过敏原,仅含一种主要过敏原(优先选择Bet v I)而非所有四种主要过敏原的提取物,就足以用于诊断和治疗目的。