Batanero Eva, Barral Patricia, Villalba Mayte, Rodríguez Rosalía
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas, Universidad Complutense, E-28040 Madrid, Spain.
Int Arch Allergy Immunol. 2002 Apr;127(4):269-75. doi: 10.1159/000057743.
Olive pollen is one of the main causes of allergy in Mediterranean countries, where it is widely distributed. One inconvenience in studying new immunotherapies for olive pollen allergy is the lack of suitable animal models. The aim of this study was to develop a murine model of IgE sensitization to Ole e 1, the major allergen of olive pollen, which mimics the immunological features of olive pollinosis in humans and to investigate the in vivo antigenicity of the recombinant form of the allergen.
BALB/c mice were sensitized by intraperitoneal administration of natural Ole e 1 (nOle e 1) and recombinant Ole e 1 (rOle e 1) in Al(OH)(3), respectively. Serum levels of specific IgE, IgG1 and IgG2a, cytokine production and the proliferative response of splenocytes after in vitro stimulation with nOle e 1 were analyzed by ELISA and flow cytometry. The binding capacity of rOle e 1-specific IgG1 was examined by ELISA and immunoblotting.
Sensitization with nOle e 1 or rOle e 1 induced high levels of specific IgE and IgG1 versus low IgG2a antibody levels. Splenocytes from sensitized mice exhibited a proliferative response to nOle e 1. In vitro stimulated splenic cells from nOle e 1-primed mice produced IL-4 and low or nondetectable levels of IFN-gamma. Specific IgE and IgG1 antibodies of immunized mice bound to the same Ole e 1 isoforms and showed a similar degree of cross-reactivity as observed for human IgE. Mouse specific nOle e 1 IgG1 was strongly inhibited by IgE from allergic patients. The IgG1 antibodies elicited by rOle e 1 reacted with both the recombinant and natural forms of the allergen.
A murine model of Ole e 1 sensitization has been established. rOle e 1 shows similar allergenicity and antigenicity to its natural form. This model should provide a useful tool for evaluating antigenic molecules and exploring new therapeutic approaches in order to treat IgE-mediated olive pollinosis.
橄榄花粉是地中海国家过敏的主要原因之一，在这些国家广泛分布。研究橄榄花粉过敏新免疫疗法的一个不便之处是缺乏合适的动物模型。本研究的目的是建立一种对橄榄花粉主要过敏原Ole e 1产生IgE致敏的小鼠模型，该模型模拟人类橄榄花粉症的免疫特征，并研究过敏原重组形式的体内抗原性。
分别通过腹腔注射天然Ole e 1（nOle e 1）和重组Ole e 1（rOle e 1）与氢氧化铝（Al(OH)(3)）致敏BALB/c小鼠。通过ELISA和流式细胞术分析用nOle e 1体外刺激后血清中特异性IgE、IgG1和IgG2a水平、细胞因子产生以及脾细胞的增殖反应。通过ELISA和免疫印迹检测rOle e 1特异性IgG1的结合能力。
用nOle e 1或rOle e 1致敏诱导产生高水平的特异性IgE和IgG1，而IgG2a抗体水平较低。致敏小鼠的脾细胞对nOle e 1表现出增殖反应。用nOle e 1预致敏小鼠的体外刺激脾细胞产生IL-4，IFN-γ水平低或检测不到。免疫小鼠的特异性IgE和IgG1抗体与相同的Ole e 1亚型结合，并且显示出与人IgE观察到的相似程度的交叉反应性。小鼠特异性nOle e 1 IgG1被过敏患者的IgE强烈抑制。rOle e 1引发的IgG1抗体与过敏原的重组形式和天然形式均发生反应。
已建立Ole e 1致敏的小鼠模型。rOle e 1与其天然形式显示出相似的致敏性和抗原性。该模型应为评估抗原分子和探索治疗IgE介导的橄榄花粉症的新治疗方法提供有用的工具。
