对酪蛋白敏感或不敏感的牛奶过敏儿童中t细胞反应的变化

Miao Qing, Li Zhen, Li Yongge, Wang Yan, Reng Yi-xin, Guan Hui, Xu Wei, Jiang Nannan, Huang Huijie, Hou Xiao-ling, Xiang Li
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The expression level of T cell related transcription factor in PBMCs or total CD4+ cells were detected by quantitative PCR, and the release of T cell related cytokines in donor’s serum was determined by Luminex xMAP 200 System. Upon in-vitro stimulation with casein protein, the proliferation rates of PBMCs-derived T cells were assessed by flow cytometry, and T related cytokines production in culture supernatant was evaluated as well.Results: A significantly higher level of GATA3 expression transcripts in PBMCs or total CD4+ cells were detected in casein-sensitized CMA patients than those without casein sensitization. An increase in IL-4 (74.1±16.5pg/ml vs 64.9±12.8 pg/ml, P<0.05) and a decrease in IL-10 (3.1±2.4 pg/ml vs 4.9±2.8 pg/ml, P<0.05) production were found in casein-sensitized CMA patient’s plasma than patients without casein sensitization. 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引用次数: 0

摘要

目的:探讨酪蛋白致敏与非酪蛋白致敏奶牛牛奶过敏(CMA)患者过敏原特异性T免疫反应的潜在差异。方法:选取2017年1 - 8月在北京儿童医院门诊就诊的ige依赖性CMA患儿25例,其中对酪蛋白成分致敏阳性的CMA患儿14例,对牛奶致敏但酪蛋白成分sIgE阴性的CMA患儿11例,另外10例无CMA的特应性对照者也作为阴性对照组。采用定量PCR检测外周血淋巴细胞或总CD4+细胞中T细胞相关转录因子的表达水平,采用Luminex xMAP 200系统检测供体血清中T细胞相关细胞因子的释放。在体外酪蛋白刺激下,流式细胞术评估pbmcs来源的T细胞的增殖率,并评估培养上清中T相关细胞因子的产生。结果:酪蛋白致敏CMA患者PBMCs或总CD4+细胞中GATA3表达转录本水平明显高于未致敏CMA患者。酪蛋白致敏CMA患者血浆中IL-4含量(74.1±16.5pg/ml vs 64.9±12.8 pg/ml, P<0.05)升高,IL-10含量(3.1±2.4 pg/ml vs 4.9±2.8 pg/ml, P<0.05)降低。乳蛋白混合物体外刺激后,酪蛋白致敏CMA患者的酪蛋白特异性CD4+T细胞增殖率(30.0±5.8% vs 20.7±5.5%,P<0.05)显著高于非酪蛋白致敏CMA患者,IL-4(4.1±13.6 vs 64.7±12.3pg/ml, P<0.05)和IL- 13(48.7±10.3 vs 38.7±9.4pg/ml, P<0.05)的产量显著高于非酪蛋白致敏CMA患者。结论:我们的研究结果初步揭示了不同致敏成分的CMA患者具有不同的T细胞介导的免疫应答。
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Variations in T-Cell Response Among Cow’s Milk Allergy Children With Or Without Sensitization to Casein Protein
Objective: To explore the potential differences in allergen-specific T immunological responses among cow’s milk allergic(CMA) patients with or without casein sensitization.Methods: A total of 25 children with IgE-dependent CMA were recruited from outpatient clinic in Beijing Children’s Hospital from January to August of 2017, including 14 CMA patient with a positive sensitization to casein component, 11 CMA patient being sensitized to cow’s milk, but a negative sIgE for casein component, and another 10 atopic control subjects without CMA were also included as negative control group in this study. The expression level of T cell related transcription factor in PBMCs or total CD4+ cells were detected by quantitative PCR, and the release of T cell related cytokines in donor’s serum was determined by Luminex xMAP 200 System. Upon in-vitro stimulation with casein protein, the proliferation rates of PBMCs-derived T cells were assessed by flow cytometry, and T related cytokines production in culture supernatant was evaluated as well.Results: A significantly higher level of GATA3 expression transcripts in PBMCs or total CD4+ cells were detected in casein-sensitized CMA patients than those without casein sensitization. An increase in IL-4 (74.1±16.5pg/ml vs 64.9±12.8 pg/ml, P<0.05) and a decrease in IL-10 (3.1±2.4 pg/ml vs 4.9±2.8 pg/ml, P<0.05) production were found in casein-sensitized CMA patient’s plasma than patients without casein sensitization. Upon in-vitro stimulation with cow’s milk protein mixture, a significantly higher proliferation rate of casein-specific CD4+T cells (30.0±5.8% vs 20.7±5.5%, P<0.05) as well as increased productions of IL-4(4.1±13.6 vs 64.7±12.3pg/ml, P<0.05) and IL- 13(48.7±10.3 vs 38.7±9.4pg/ml, P<0.05) were found in casein-sensitized CMA patients compared those without casein sensitization.Conclusions: Our findings preliminarily revealed that a distinctive T cell mediated immune response among CMA patient with the different sensitized component.
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