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目的比较人和BALB/c小鼠对梭子蟹过敏的组分差异,评价用小鼠构建过敏模型对研究人类过敏症的可靠性,并对主要过敏原进行分离纯化。方法制备梭子蟹蛋白浸液,经皮下注射建立小鼠过敏模型,获得特异的小鼠IgE(sIgE)血清,同时收集蟹过敏患者血清,采用Western blot法鉴定蟹的过敏组分,通过硫酸铵分段盐析和DEAE离子交换层析对主要过敏组分进行分离纯化。结果用梭子蟹蛋白浸液成功建立了BALB/c小鼠过敏模型,并获得了高效价的sIgE血清。梭子蟹中有一相对分子质量为89 000的蛋白组分为人和小鼠共有的主要过敏原,通过硫酸铵分段盐析法和DEAE离子交换层析法分离到了该组分。结论小鼠和人对梭子蟹过敏的组分基本一致,提示小鼠过敏模型可成为人类过敏症研究的可靠动物模型,纯化单一的过敏原组分有望提高过敏症诊断和治疗的特异性。
OBJECTIVE: To compare the component differences between human and BALB / c mice on the allergicus of Portunus trituberculatus, and to evaluate the reliability of the mouse allergy model in the study of human allergy and to isolate and purify the major allergens. Methods The mouse crab infusions were prepared and the mouse allergic model was established by subcutaneous injection. The specific mouse IgE (sIgE) serum was collected. At the same time, the serum of crab allergy sufferers was collected. The allergic components of crab were identified by Western blot. Salting out and DEAE ion exchange chromatography were used to separate and purify the main allergy components. Results BALB / c mouse model of allergy was established successfully by the shuttle of crab protein and sIgE serum with high titer was obtained. One of the major allergens shared by humans and mice is a protein component with a relative molecular mass of 89,000 in the shuttle pool, which was isolated by ammonium sulfate fractional salting-out and DEAE ion exchange chromatography. Conclusions The mice and humans are basically identical in allergy to B. crab, suggesting that the mouse allergy model may be a reliable animal model for human allergy research. Purifying a single allergen component is expected to improve the specificity of diagnosis and treatment for allergy.