Quiralte J, González E, Arias De Saavedra J M, Villalba M, Florido J F, Sáenz De San Pedro B, Rodríguez R
Unidad de Alergia, Hospital Ciudad de Jaén, Jaén, España.
Int Arch Allergy Immunol. 2000 Jun;122(2):101-7. doi: 10.1159/000024365.
Recombinant allergens have potential advantages over conventional allergenic extracts. However, these recombinant allergens should be evaluated for their antigenic activity and compared with their natural counterparts before being used for clinical purposes.
We studied 33 patients with seasonal rhinitis and/or bronchial asthma and a positive skin prick test to Olea europaea pollen extract, 10 atopic patients with no history of pollinosis and a negative skin prick test to O. europaea extract and 10 healthy controls. Skin prick tests and determination by ELISA of specific IgE to natural Ole e 1 (nOle e 1) and recombinant Ole e 1 (rOle e 1) expressed in Pichia pastoris were performed in all patients and controls. Inhibition assays were performed between nOle e 1 and rOle e 1 by ELISA.
All patients with O. europaea pollinosis had positive skin test responses to both commercial O. europaea extract and nOle e 1 allergen, and all reacted to rOle e 1 on the skin prick test. The nonatopic and atopic control subjects with negative olive pollen skin test results did not react to rOle e 1 on the skin prick test, even at the highest concentrations, confirming the specificity of this test. We found a weak correlation between the wheal surface area produced by the prick test with nOle e 1 and the wheal surface area produced by rOle e 1 at 10 microgram/ml (r = 0.42, p < 0.05). Comparison of specific IgE against both nOle e 1 and rOle e 1 in the patients did not reveal any significant difference. There was a strong correlation between the amount of specific IgE against nOle e 1 and rOle e 1 (r = 0.99, p < 0.01). The two proteins displayed the same extent of binding inhibition to IgE antibodies in ELISA inhibition experiments.
These results confirm the immunological activity of rOle e 1 expressed in P. pastoris and indicate that Ole e 1 is one of the major allergens in O. europaea pollinosis as evaluated by skin prick test and serological methods. The correlation between rOle e 1 and nOle e 1 in skin test results and serologic data indicates the potential of recombinant allergens for clinical applications and diagnosis of O. europaea pollen allergy.
重组变应原相较于传统变应原提取物具有潜在优势。然而,在将这些重组变应原用于临床之前,应对其抗原活性进行评估,并与天然对应物进行比较。
我们研究了33例季节性鼻炎和/或支气管哮喘患者,他们对油橄榄花粉提取物皮肤点刺试验呈阳性,10例无花粉症病史且对油橄榄提取物皮肤点刺试验呈阴性的特应性患者,以及10名健康对照者。对所有患者和对照者进行了皮肤点刺试验,并通过酶联免疫吸附测定(ELISA)法测定了针对天然油橄榄1(nOle e 1)和在毕赤酵母中表达的重组油橄榄1(rOle e 1)的特异性IgE。通过ELISA法在nOle e 1和rOle e 1之间进行抑制试验。
所有油橄榄花粉症患者对市售油橄榄提取物和nOle e 1变应原的皮肤试验反应均为阳性,且在皮肤点刺试验中对rOle e 1均有反应。皮肤试验结果为阴性的非特应性和特应性对照受试者,即使在最高浓度下,对rOle e 1的皮肤点刺试验也无反应,证实了该试验的特异性。我们发现,在10微克/毫升时,nOle e 1点刺试验产生的风团表面积与rOle e 1产生的风团表面积之间存在弱相关性(r = 0.42,p < 0.05)。患者中针对nOle e 1和rOle e 1的特异性IgE比较未显示出任何显著差异。针对nOle e 1和rOle e 1的特异性IgE量之间存在强相关性(r = 0.99,p < 0.01)。在ELISA抑制试验中,这两种蛋白质对IgE抗体的结合抑制程度相同。
这些结果证实了在毕赤酵母中表达的rOle e 1的免疫活性,并表明通过皮肤点刺试验和血清学方法评估,油橄榄1是油橄榄花粉症的主要变应原之一。rOle e 1与nOle e 1在皮肤试验结果和血清学数据中的相关性表明重组变应原在油橄榄花粉过敏临床应用和诊断中的潜力。