纯化的天然和重组猫源Fel d 1及猫白蛋白在猫过敏体外诊断中的应用
Purified natural and recombinant Fel d 1 and cat albumin in in vitro diagnostics for cat allergy.
作者信息
van Ree R, van Leeuwen W A, Bulder I, Bond J, Aalberse R C
机构信息
Central Laboratory of the Netherlands Red Cross Blood Transfusion Service and the Laboratory for Experimental and Clinical Immunology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands.
出版信息
J Allergy Clin Immunol. 1999 Dec;104(6):1223-30. doi: 10.1016/s0091-6749(99)70017-5.
BACKGROUND
Current diagnostics and therapeutics for cat allergy are based on cat epithelial extracts originating from highly variable source materials. This gives rise to several problems: variability of allergen composition, contamination with house dust mite allergens, and potential transfer of pathogenic agents.
OBJECTIVE
The aim of this study was to investigate the feasibility of replacing cat epithelial extracts with purified natural or recombinant allergens.
METHODS
Sera (n = 509) were selected on the basis of a positive cat RAST result and tested in a RAST for IgE reactivity to purified Fel d 1, cat albumin (CA), or both. The analysis was performed with both natural and recombinant allergens. In addition, some sera were further analyzed by means of immunoblotting. A serum pool was used for cat RAST inhibition with purified natural and recombinant allergens as inhibitors.
RESULTS
Natural and recombinant Fel d 1 caused very similar results: 94.1% and 96.1% positive test results, respectively. In general, the negative sera were low responders to cat extract. The addition of CA (16.7% positive sera) resulted in a decrease in the number of discrepencies between purified allergens and whole extract to 2.8%. Only for 2% of all sera, sensitization to cat was largely explained by IgE reactivity to CA. IgE reactivity to Fel d 1 accounts for 88% of the total IgE response to cat allergens, as was demonstrated by RAST, with Fel d 1 concentrations nearing saturation. Recombinant Fel d 1 performed equally well in the RAST analysis. Recombinant CA was succesfully expressed in the yeast Pichia pastoris, and its immune reactivity closely resembled that of its natural counterpart.
CONCLUSION
Natural and recombinant Fel d 1 and CA are good candidates for replacing ill-defined cat dander extracts in diagnostics for cat allergy. Although CA is not essential for the vast majority of cat-sensitized patients, some subjects are selectively sensitized to this serum protein.
背景
目前猫过敏的诊断和治疗方法基于源自高度可变来源材料的猫上皮提取物。这引发了几个问题：过敏原成分的变异性、屋尘螨过敏原的污染以及病原体的潜在传播。
目的
本研究的目的是探讨用纯化的天然或重组过敏原替代猫上皮提取物的可行性。
方法
根据猫RAST结果为阳性选择血清（n = 509），并在RAST中检测其对纯化的猫过敏原1（Fel d 1）、猫白蛋白（CA）或两者的IgE反应性。使用天然和重组过敏原进行分析。此外，一些血清通过免疫印迹进一步分析。使用纯化的天然和重组过敏原作为抑制剂，用血清池进行猫RAST抑制试验。
结果
天然和重组的Fel d 1产生了非常相似的结果：阳性检测结果分别为94.1%和96.1%。一般来说，阴性血清对猫提取物的反应较低。添加CA（16.7%阳性血清）后，纯化过敏原与全提取物之间的差异数量减少至2.8%。仅2%的所有血清中，对猫的致敏主要由对CA的IgE反应性解释。如RAST所示，对Fel d 1的IgE反应性占对猫过敏原总IgE反应的88%，此时Fel d 1浓度接近饱和。重组Fel d 1在RAST分析中表现同样良好。重组CA在毕赤酵母中成功表达，其免疫反应性与其天然对应物非常相似。
结论
天然和重组的Fel d 1和CA是在猫过敏诊断中替代不明确的猫皮屑提取物的良好候选物。尽管CA对绝大多数猫致敏患者不是必需的，但一些受试者对这种血清蛋白有选择性致敏。
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