Paranos S, Pravica V, Bonaci B, Stojković-Mostarica M
Department of Allergy and Clinical Immunology, Zvezdara University Medical Centre, Belgrade.
Srp Arh Celok Lek. 1998 Mar-Apr;126(3-4):92-6.
The recently published data suggest that atopic patients have an enhanced ability to produce IL-4, even in response to antigens other than common environmental allergens or helminthic components. This aberrant IL-4 production by Th cells may be one of the immune alterations encoded by nonMHC genes in the control of basal IgE level [1-7]. The existence of atopen-CD4+ T cell clones that have diverse repertoires might be relevant for aetiopathogenesis of atopic diseases [8-10]. Being subjected continuously to the environmental antigens (allergens), the immune system, especially T cells, is permanently activated and stimulated in response to that challenge. Due to that, substantial amount of diverse cytokines (and their soluble receptors) could be found in the serum. Accordingly, it could be expected to find raised serum IL-4 concentration in atopic persons. In this study we investigated concentrations of IL-4, total IgE and allergen-specific IgE in sera of atopic persons susceptible to pollen allergens. The main goal was to estimate whether the immunologic profile of atopics is defined by increased serum concentration of IL-4 apart from total and allergen-specific serum IgE.
Patients. We selected 9 atopic patients in the range of 19-35 years, susceptible to grass or weed pollens and suffering from allergic rhinoconjuctivitis. There were 6 females and 3 males (mean age 31.2 years). Atopic allergy was proven by clinical history, skin prick test and by means of in vitro test (positive serum allergen-specific IgE antibodies, RAST/radioallergosorbent tests [11]. As controls we randomly selected B blood donors (6 females, 2 males, mean age 31.3). Study design. The study was carried out from August till September 1995. None of selected patients had previously accomplished allergen-specific immunotherapy nor had been medicated by corticosteroids (topic or systemic) [12-15]. Selected persons were not allowed to take medications such as antihistamines, beta-2 agonists or tricyclic antidepressants at least 10 days prior to the study. We performed in vitro tests for determining concentrations of IL-4, total and pollen-specific serum IgE. Sera were sampled in the morning and were stored frozen at -20 degrees C until analysis. Determination of IL-4, total and allergen specific IgE in serum. Interleukin-4 measurement was carried out in blind fashion with an ELISA kit (Intertest-4 ELISA, Genzyme, Cambridge) according to the manufacturer's instructions. A reference curve was obtained by plotting the IL-4 concentration of several standard dilutions versus an absorbency. The determination limit was 0.045 pg/ml. For determining total serum IgE we used commercially available enzyme immunoassay (EIA Phadezym IgE PRIST, Pharmacia, Uppsala). Normal values was up to 120 IU/L (international unit per ml). Allergen specific IgE was determined by semiquantitative EIA method (RAST Phadezym, Pharmacia, Uppsala) and expressed in Phadebas RAST Unit (PRU) according to standard curve done by manufacturer. Study was carried out after approval of the Ethic Committee of our Hospital and with the obtained patients consents. For statistical analysis we used nonparametric tests (U-test and the Spearman rank correlation). For all comparisons, statistical significance was considered to be present if p < 0.05.
Among selected patients, 4 persons had high concentration of serum IgE antibodies (RAST class 4) against grass pollens, two persons had IgE-specific (RAST, class 4) to weed pollens and 2 patients had IgE antibodies (RAST class 4) against weed and grass pollens simultaneously. RAST of class 3 was registered in the serum of one person. Registered serum total IgE, allergen-specifc IgE and IL-4 are shown in Table 1. Serum IL-4 was significantly higher in atopics (U-test, SR 43.53, p.05) as well as total IgE (ER 493, p.05). In atopics, IL-4 was not in correlation with either total or allergen-specific IgE (IL-4 versus total IgE, p = +0.190;
最近发表的数据表明，特应性患者产生白细胞介素-4（IL-4）的能力增强，即使是对常见环境过敏原或蠕虫成分以外的抗原作出反应时也是如此。辅助性T细胞这种异常的IL-4产生可能是非主要组织相容性复合体（nonMHC）基因在控制基础免疫球蛋白E（IgE）水平时所编码的免疫改变之一[1-7]。具有不同谱系的特应性CD4 + T细胞克隆的存在可能与特应性疾病的病因发病机制有关[8-10]。由于持续接触环境抗原（过敏原），免疫系统，尤其是T细胞，会因应对这种挑战而持续被激活和刺激。因此，血清中可发现大量不同的细胞因子（及其可溶性受体）。相应地，可以预期在特应性个体中发现血清IL-4浓度升高。在本研究中，我们调查了对花粉过敏原敏感的特应性个体血清中IL-4、总IgE和过敏原特异性IgE的浓度。主要目的是评估除了总血清IgE和过敏原特异性血清IgE外，特应性个体的免疫特征是否由血清IL-4浓度升高所定义。
患者。我们选择了9名年龄在19至35岁之间、对草或杂草花粉敏感且患有过敏性鼻结膜炎的特应性患者。其中有6名女性和3名男性（平均年龄31.2岁）。特应性过敏通过临床病史、皮肤点刺试验以及体外试验（血清过敏原特异性IgE抗体阳性，放射性变应原吸附试验/RAST）得以证实[11]。作为对照，我们随机选择了8名献血者（6名女性、2名男性，平均年龄31.3岁）。研究设计。该研究于1995年8月至9月进行。所选患者此前均未完成过敏原特异性免疫治疗，也未接受过皮质类固醇药物（局部或全身用药）治疗[12-15]。所选人员在研究前至少10天不允许服用抗组胺药、β-2激动剂或三环类抗抑郁药。我们进行了体外试验以测定IL-4、总血清IgE和花粉特异性血清IgE的浓度。血清于早晨采集，并在-20℃下冷冻保存直至分析。血清中IL-4、总IgE和过敏原特异性IgE的测定。使用ELISA试剂盒（Intertest-4 ELISA，Genzyme，剑桥）按照制造商的说明以盲法进行白细胞介素-4的测量。通过绘制几种标准稀释液的IL-4浓度与吸光度的关系获得参考曲线。测定下限为0.045 pg/ml。为了测定总血清IgE，我们使用了市售的酶免疫测定法（EIA Phadezym IgE PRIST，Pharmacia，乌普萨拉）。正常值高达120 IU/L（每毫升国际单位）。过敏原特异性IgE通过半定量EIA方法（RAST Phadezym，Pharmacia，乌普萨拉）测定，并根据制造商绘制的标准曲线以Phadebas RAST单位（PRU）表示。本研究在获得我院伦理委员会批准并取得患者同意后进行。对于统计分析，我们使用了非参数检验（U检验和Spearman等级相关性检验）。对于所有比较，如果p < 0.05，则认为具有统计学意义。
在所选患者中，4人针对草花粉的血清IgE抗体浓度较高（RAST 4级），2人对杂草花粉具有IgE特异性（RAST，4级），2名患者同时对杂草和草花粉具有IgE抗体（RAST 4级）。一人血清中记录到RAST 3级。表1显示了记录的血清总IgE、过敏原特异性IgE和IL-4。特应性个体的血清IL-4显著更高（U检验，SR 43.53，p.05），总IgE也是如此（ER 493，p.05）。在特应性个体中，IL-4与总IgE或过敏原特异性IgE均无相关性（IL-4与总IgE相比，p = +0.190；
（注：原文中“SR 43.53”“ER 493”表述似乎不太准确，可能是记录有误，但按要求未作修改。）
