Scott-Taylor Tim H, Axinia Stefan-Claudiu, Strobel Stephan
Faculty of Life Sciences and Computing, London Metropolitan University, London, United Kingdom.
Faculty of Life Sciences and Computing, London Metropolitan University, London, United Kingdom.
Ann Allergy Asthma Immunol. 2017 Sep;119(3):274-283. doi: 10.1016/j.anai.2017.06.015.
Peripheral blood mononuclear cells (PBMCs) proliferate inconsistently, rendering current lymphoproliferation assays unreliable in diagnosis.
To investigate the utility and nature of proliferation responses in allergy by comparison of the standard lymphoproliferation with a new dendritic cell (DC) stimulated assay.
Monocyte-derived DCs were pulsed with allergens and incubated with autologous T cells for 7 days. DC-stimulated and standard PBMC proliferation responses to 3 common dietary allergens in children with allergy and without atopy were measured by incorporation of tritiated thymidine and reduction of carboxyl fluorescein succinimidyl ester staining.
The DC presentation of sensitizing allergens induced significantly higher proliferative responses than PBMC stimulation (P = .04) and greater distinction between normal and allergic responses. DC-induced stimulation indices of children without sensitivity and those with allergy were significantly different with all 3 foods (P < .001). All children with allergy presented with peanut allergy and 12 of 14 (86%) β-lactoglobulin-pulsed DC preparations proliferated more than 3.3-fold above un-pulsed cells, but 8 of 18 children (44%) with ovalbumin egg allergy showed proliferation below this level. The stimulation index of DC tritiated thymidine incorporation correlated closely with carboxyl fluorescein succinimidyl ester reduction (P < .001). Sensitivity of detection of peanut, milk, or egg allergy was 100%, 85.7%, or 55.6% and specificity was 60%, 88.9%, or 86.7%, respectively. DC-stimulated T cells expressed increased levels of CD45 RO and CD25 and most produced interferon-γ. DC-stimulated proliferation correlated with total immunoglobulin E and peanut antigen-stimulated proliferation correlated with peanut specific immunoglobulin E (P = .03).
The DC-induced lymphoproliferation had higher sensitivity, specificity, and reproducibility than the standard assay and caused increased memory and activated T-cell proliferation in children with food allergy.
外周血单个核细胞（PBMCs）增殖不一致，使得当前的淋巴细胞增殖检测在诊断中不可靠。
通过将标准淋巴细胞增殖检测与一种新的树突状细胞（DC）刺激检测进行比较，研究过敏中增殖反应的效用和性质。
用过敏原刺激单核细胞衍生的DCs，并与自体T细胞孵育7天。通过掺入氚标记的胸腺嘧啶核苷和羧基荧光素琥珀酰亚胺酯染色的减少，测量过敏儿童和非特应性儿童对3种常见食物过敏原的DC刺激和标准PBMC增殖反应。
致敏过敏原的DC呈递诱导的增殖反应明显高于PBMC刺激（P = 0.04），并且正常反应和过敏反应之间的差异更大。对于所有3种食物，无敏感性儿童和过敏儿童的DC诱导刺激指数有显著差异（P < 0.001）。所有过敏儿童均表现为花生过敏，14份（86%）经β-乳球蛋白刺激的DC制剂中有12份增殖超过未刺激细胞的3.3倍，但18名卵清蛋白过敏儿童中有8名（44%）的增殖低于该水平。DC氚标记胸腺嘧啶核苷掺入的刺激指数与羧基荧光素琥珀酰亚胺酯减少密切相关（P < 0.001）。花生、牛奶或鸡蛋过敏检测的敏感性分别为100%、85.7%或55.6%，特异性分别为60%、88.9%或86.
