Jutel M, Wyss-Coray T, Carballido J M, Blaser K, Müller U R, Pichler W J
Zieglerspital, Bern, Switzerland.
Clin Exp Allergy. 1995 Nov;25(11):1108-17. doi: 10.1111/j.1365-2222.1995.tb03258.x.
The synthesis of IgE is regulated by cytokines secreted from T-helper cells. The studies on cytokine secretion by peripheral blood mononuclear cells (PBMC) upon stimulation with antigen or allergen are difficult due to low levels of cytokines, especially of interleukin-4 (IL-4).
In this study we tried to establish a culture system, which could enable the measurement of the cytokine profiles in specifically activated cultures.
Three methods to potentiate cytokine secretion were evaluated: PBMC from bee venom or house dust mite (Dermatophagoides pteronyssinus) allergic patients as well as normal subjects were stimulated either with the major bee venom allergen phospholipase A2 (PLA) or with the major D. pteronyssinus allergen (Der p 1) or with the control antigens tetanus toxoid (TT) and purified protein derivate (PPD). After 7 days of culture the cells were restimulated either with plastic bound OKT3 F(ab)2 monoclonal antibodies (MoAbs), with the appropriate antigen + antigen presenting cells or with IL-2. The secretion of cytokines (IL-4, IFN gamma) was measured after restimulation of the cultures (day 8).
While OKT3 F(ab)2 was unable to activate resting T cells, it could restimulate preactivated cells. Restimulation with OKT3 F(ab)2 induced higher IL-4 and IFN gamma secretion than restimulation with IL-2 or antigen. TT and PLA stimulated a similar cytokine secretion profile in normal and PLA allergic subjects with substantial levels of both IL-4 and IFN gamma. In contrast, PPD induced virtually only IFN gamma secretion. Der p 1 stimulated mainly IL-4 secretion but also IFN gamma production in some mite allergic patients.
We have established a cell culture system, which combines antigen specificity with a strong cytokine inducing signal provided by anti-CD3 MoAbs. TH-1 and TH-2 characteristic cytokine patterns can be observed in short-term PBMC cultures already after 8 days of culture.
IgE的合成受T辅助细胞分泌的细胞因子调控。由于细胞因子水平较低，尤其是白细胞介素-4（IL-4），因此研究外周血单个核细胞（PBMC）在抗原或变应原刺激下的细胞因子分泌较为困难。
在本研究中，我们试图建立一种培养系统，该系统能够在特异性激活的培养物中测量细胞因子谱。
评估了三种增强细胞因子分泌的方法：用主要的蜂毒变应原磷脂酶A2（PLA）、主要的屋尘螨变应原（Der p 1）或对照抗原破伤风类毒素（TT）和纯化蛋白衍生物（PPD）刺激来自蜂毒或屋尘螨（粉尘螨）过敏患者以及正常受试者的PBMC。培养7天后，用塑料包被的OKT3 F(ab)2单克隆抗体（MoAbs）、适当的抗原+抗原呈递细胞或IL-2对细胞进行再刺激。在培养物再刺激后（第8天）测量细胞因子（IL-4、IFNγ）的分泌。
虽然OKT3 F(ab)2不能激活静息T细胞，但它可以再刺激预激活的细胞。用OKT3 F(ab)2再刺激诱导的IL-4和IFNγ分泌高于用IL-2或抗原再刺激。TT和PLA在正常和PLA过敏受试者中刺激了相似的细胞因子分泌谱，IL-4和IFNγ水平都很高。相比之下，PPD实际上只诱导IFNγ分泌。Der p 1主要刺激一些螨过敏患者的IL-4分泌，但也刺激IFNγ产生。
我们建立了一种细胞培养系统，该系统将抗原特异性与抗CD3 MoAbs提供的强大细胞因子诱导信号相结合。在培养8天后的短期PBMC培养物中已经可以观察到TH-1和TH-2特征性细胞因子模式。
