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目的探讨质粒转染技术人工合成鸡蛋组分蛋白的免疫原性;鸡蛋组分蛋白与过敏患者鸡蛋特异性免疫球蛋白(immunoglobulin E,Ig E)含量及皮肤试验的相关性。方法通过质粒转染技术人工合成鸡蛋组分蛋白,并用Western blot方法验证目的蛋白免疫原性。再将待检测组分蛋白和鸡蛋过敏患者血清中特异性Ig E结合,分析各蛋白组分对鸡蛋过敏的临床诊断价值。结果人工合成的鸡蛋组分蛋白(卵类黏蛋白、卵白蛋白、卵转铁蛋白、溶菌酶)检测后仍能表现出较好的免疫原性;鸡蛋特异性Ig E>10 k U/L组的患儿鸡蛋过敏原点刺阳性率明显高于<10 k U/L组(95.5%vs.57.8%,P=0.000 3);卵类黏蛋白光密度值(optical density,OD)450 nm=0.81和卵白蛋白OD450 nm=0.567为鸡蛋过敏原的主要成分;鸡蛋特异性Ig E(>10 k U/L)含量高的患儿血清中卵白蛋白抗体含量较高,且鸡蛋特异性Ig E含量(>10 k U/L)与血清卵白蛋白抗体含量呈明显正相关关系(p=0.630 9,P=0.002 2)。而鸡蛋特异性Ig E<10 k U/L的患儿血清中卵类黏蛋白抗体含量高。鸡蛋特异性Ig E含量(<10 k U/L)与血清卵类黏蛋白抗体含量也呈明显正相关关系(p=0.756 0,P<0.000 1)。结论通过人工合成质粒转染的鸡蛋组分蛋白具有较好的免疫原性,对今后食物过敏原的纯化有一定价值;鸡蛋组分蛋白对儿童鸡蛋过敏的诊断有重要价值。
OBJECTIVE: To investigate the immunogenicity of synthetic protein components of egg by plasmid transfection and the correlation between egg-specific immunoglobulin E (IgE) content and skin test in allergic patients. Methods The protein components of eggs were synthesized by plasmid transfection and the immunogenicity of the target protein was verified by Western blot. The protein to be tested and the specific IgE in the serum of egg allergic patients were combined to analyze the clinical diagnostic value of each protein component on egg allergy. Results The synthetic protein components (ovomucin, ovalbumin, egg transferrin and lysozyme) still showed good immunogenicity after being tested. The egg-specific Ig E> 10 kU / L group (95.5% vs.57.8%, P = 0.0003). The optical density (OD) of egg mucins 450 nm was 0.81 And ovalbumin (OD450 nm = 0.567) were the major components of egg allergen. The content of egg albumin antibody in serum of children with high egg-specific Ig E (> 10 kU / L) was high, and the egg-specific Ig E content > 10 kU / L) was positively correlated with serum albumin antibody (p = 0.6309, P = 0.002 2). The egg-specific Ig E <10 kU / L in children with serum levels of egg mucin antibodies. Egg-specific IgE levels (<10 kU / L) were also positively correlated with serum egg-like mucin antibody levels (p = 0.756 0, P <0.0001). Conclusion The protein of egg components transfected by artificial plasmids has good immunogenicity and has some value for the future purification of food allergens. The protein of egg components is of great value in the diagnosis of egg allergy in children.