Gołebiowska-Wawrzyniak Maria, Markiewicz Katarzyna, Rytwiński Krzysztof, Buława Ewa, Czerwińska Iwona, Wnuk Aleksander, Derentowicz Piotr, Kozar Agata, Siwińska-Gołebiowska Henryka
Zakład Immunologii Klinicznej, Instytut Matki i Dziecka, Kasprzaka 17a, 01-211 Warszawa, Poland.
Med Wieku Rozwoj. 2003 Oct-Dec;7(4 Pt 2):565-75.
As the frequency of allergic disease and especially the food allergy in children is still increasing - we undertook studies to determine the specific sensitivity of lymphocytes T in order to improve the diagnostics and monitoring of treatment. The usefulness of evaluation CD69 and PCNA expression of T cells from children with cow's milk allergy (CMA) in these procedures was examined.
The study on usefulness of lymphocytes CD69 and PCNA detection for diagnosis of CMA in children and for monitoring of the treatment.
The subjects in this study included 26 children from 1.5 to 15 years old with CMA who were under clinical care of our Immunological Department. Patients were examined (medical history of the child, physical examination, specific food provocation) and laboratory investigations were made before therapy and after 6 months of elimination diet. Allergen-specific IgE antibody was determined. Children's T cell cultures were stimulated by beta-lactoglobulin, alpha-casein, whole cow's milk, phytohaemoglutinin PHA and without stimulation. Expression of T cells CD69 and PCNA from patients with CMA and 10 healthy children were analysed by use of flow cytometry. 10 children with CMA after 6 months of elimination diet and next 2 weeks with cow's milk provocation were examined also for CD69 and PCNA expression in lymphocytes from peripheral blood.
After 6 months of elimination diet it seems that the expression of CD69 receptor has tendency to fall, these differences, however, are not statistically significant. But after next 2 weeks with cow's milk diet the expression of CD69 was increased with statistically significance (p=0.04). T cells PCNA expression of children with CMA was lower than in healthy children in PHA stimulated cultures. Our study indicates that PCNA expression of T cells from patients with CMA increased even more after 6 months of elimination diet in all cultures but there was statistically significant differences only after stimulation with cow's milk and PHA (p=0.05 and p=0.04 - respectively). Subsequently through 2 weeks children were provoked with cow's milk and after that PCNA expression in vivo lowered (no statistically significant differences).
CD69 expression of T lymphocytes was significantly higher after specific stimulation of children with cow's milk allergy (p=0.04). The evaluation of CD69 expression of children with CMA may be also useful in monitoring of treatment. Estimation of PCNA expression suggests the efficiency of T cells connected with DNA synthesis related to the stage of allergy disease and demands further investigations.
由于儿童过敏性疾病尤其是食物过敏的发病率仍在上升，我们开展了研究以确定T淋巴细胞的特异性敏感性，从而改善诊断和治疗监测。研究了评估牛奶过敏（CMA）儿童T细胞CD69和PCNA表达在这些过程中的有用性。
研究淋巴细胞CD69和PCNA检测对儿童CMA诊断及治疗监测的有用性。
本研究的受试者包括26名年龄在1.5至15岁之间患有CMA且在我们免疫科接受临床护理的儿童。在治疗前和进行6个月的排除饮食后对患者进行检查（儿童病史、体格检查、特定食物激发试验）并进行实验室检查。测定过敏原特异性IgE抗体。用β-乳球蛋白、α-酪蛋白、全脂牛奶、植物血凝素PHA刺激儿童T细胞培养物，且有未刺激的培养物。通过流式细胞术分析CMA患者和10名健康儿童的T细胞CD69和PCNA表达。对10名经过6个月排除饮食且接下来2周进行牛奶激发试验的CMA儿童，也检测其外周血淋巴细胞中CD69和PCNA的表达。
经过6个月的排除饮食后，CD69受体的表达似乎有下降趋势，然而这些差异无统计学意义。但在接下来2周的牛奶饮食后，CD69的表达有统计学意义的增加（p = 0.04）。在PHA刺激的培养物中，CMA儿童的T细胞PCNA表达低于健康儿童。我们的研究表明，在所有培养物中，经过6个月的排除饮食后，CMA患者T细胞的PCNA表达增加更多，但仅在用牛奶和PHA刺激后才有统计学意义的差异（分别为p = 0.05和p = 0.04）。随后，对儿童进行2周的牛奶激发试验后，体内PCNA表达降低（无统计学意义的差异）。
牛奶过敏儿童经特异性刺激后，T淋巴细胞的CD69表达显著升高（p = 0.04）。评估CMA儿童CD69的表达在治疗监测中可能也有用。PCNA表达的评估提示与DNA合成相关的T细胞效率与过敏疾病阶段有关，需要进一步研究。
