Upham J W, Holt B J, Baron-Hay M J, Yabuhara A, Hales B J, Thomas W R, Loh R K, O'Keeffe P T, Palmer L, Le Souef P N
Telethon Institute for Child Health Research, University of Western Australia, Perth.
Clin Exp Allergy. 1995 Jul;25(7):634-42. doi: 10.1111/j.1365-2222.1995.tb01111.x.
It is widely held that in vitro T cell responses to allergens are more prominent in atopic than in normal individuals, though this conclusion is based upon culture techniques which fail to detect proliferative responses in a significant minority of atopics and many normals.
Study allergen-specific proliferative responses of T cells cultured in serum-free medium (SFM). Examine associations between atopic status, age and T cell reactivity.
Initially, peripheral blood mononuclear cells were stimulated with allergens or antigens in SFM, and compared with cells cultured in RPMI + 10% fetal calf serum or human AB serum. Subsequently, T cell reactivity was studied in 34 adults (20-49 years), 27 children (2-13 years), and 19 infants (< or = 10 weeks) using SFM alone.
Compared with serum-supplemented medium, SFM enhanced net T cell proliferation, both in bulk culture and when cloning at limiting dilution. In many subjects, SFM unmasked T cell reactivity to allergens which was not otherwise evident, and lowered the threshold allergen levels required for in vitro T cell triggering. For most allergens, T cell proliferative responses did not differ between adults who had specific IgE, and those who did not. The most vigorous responses observed were to ubiquitous inhalant allergens, which stimulated T cells from close to 100% of adults and children, and over 60% of infants. In contrast, responses to the 'vaccine' antigen tetanus toxoid were completely absent in the latter age group, but present in the majority of adults and children.
These findings suggest that the extent of active T cell recognition of environmental allergens has been hitherto underestimated, and further that these responses may frequently be initiated in very early life. Additionally, these findings reinforce the notion that qualitative (as opposed to quantitative) variations in specific T cell reactivity ultimately determine allergen responder phenotype.
人们普遍认为,体外T细胞对变应原的反应在特应性个体中比正常个体更为显著,尽管这一结论是基于培养技术得出的,而这些技术未能检测到相当一部分特应性个体和许多正常个体中的增殖反应。
研究在无血清培养基(SFM)中培养的T细胞的变应原特异性增殖反应。研究特应性状态、年龄与T细胞反应性之间的关联。
首先,用变应原或抗原在SFM中刺激外周血单个核细胞,并与在RPMI + 10%胎牛血清或人AB血清中培养的细胞进行比较。随后,仅使用SFM研究了34名成年人(20 - 49岁)、27名儿童(2 - 13岁)和19名婴儿(≤10周)的T细胞反应性。
与补充血清的培养基相比,SFM在大量培养和有限稀释克隆时均增强了T细胞的净增殖。在许多受试者中,SFM揭示了T细胞对变应原的反应性,而这种反应性在其他情况下并不明显,并降低了体外T细胞触发所需的变应原阈值水平。对于大多数变应原,有特异性IgE的成年人和没有特异性IgE的成年人之间的T细胞增殖反应没有差异。观察到的最强烈反应是针对普遍存在的吸入性变应原,这些变应原刺激了近100%的成年人和儿童以及超过60%的婴儿的T细胞。相比之下,后一年龄组对“疫苗”抗原破伤风类毒素完全没有反应,但在大多数成年人和儿童中存在反应。
这些发现表明,迄今为止,环境变应原的活性T细胞识别程度被低估了,而且进一步表明这些反应可能经常在生命的很早阶段就开始了。此外,这些发现强化了这样一种观念,即特异性T细胞反应性的定性(而非定量)变化最终决定了变应原反应表型。