Kulis Michael, Pons Laurent, Burks A Wesley
Department of Pediatrics, Division of Pediatric Allergy and Immunology, Duke University Medical Center, Durham, NC 27710, USA.
Int Arch Allergy Immunol. 2009;148(2):109-17. doi: 10.1159/000155741. Epub 2008 Sep 19.
Examination of IgE cross-reactivity among nuts has been limited to in vitro experiments. Cross-reactivity studies of nuts at the T cell level are difficult to interpret because of the inability to determine which cellular responses are from a true sensitization and which are due to cross-reactivity. Using a mouse model in which the sensitizing nuts are controlled may provide novel methods to investigate in vivo and T cell cross-reactivity.
C3H/HeJ mice were sensitized by intraperitoneal injection of cashew alone (monosensitized mice), or cashew plus walnut, utilizing alum as an adjuvant. Both groups underwent challenges to cashew, walnut and peanut, with subsequent monitoring of anaphylactic reactions. Anaphylactic antibodies were quantified by ELISA, and protein allergens were identified by Western blotting. Cellular responses were studied via splenocyte proliferation assay and measurement of secreted cytokines.
The monosensitized mice reacted to cashew and walnut during challenges, with significantly weaker reactions induced on challenge with peanut. Cross-reactive IgE to walnut and peanut were detected by ELISA, and the cross-reactive allergens were identified as vicilin proteins. In cellular assays, splenocytes from the monosensitized mice proliferated and produced IL-4 and IL-5 in response to cashew, walnut and peanut. The cashew- plus walnut-sensitized mice experienced stronger clinical reactions to walnut, recognized additional walnut allergens and secreted significantly more IL-4 and IL-5 in walnut-stimulated splenocyte assays compared to the monosensitized mice.
Cross-reactivity in vivo was found between cashew and walnut, while cross-reactivity among cashew, walnut and peanut was demonstrated at the T cell level.
对于坚果之间IgE交叉反应性的研究仅限于体外实验。由于无法确定哪些细胞反应来自真正的致敏作用,哪些是由交叉反应性引起的,因此在T细胞水平上对坚果的交叉反应性研究难以解释。使用一种致敏坚果得到控制的小鼠模型可能会为研究体内和T细胞交叉反应性提供新方法。
通过腹腔注射单独的腰果(单致敏小鼠)或腰果加核桃,利用明矾作为佐剂使C3H/HeJ小鼠致敏。两组小鼠均接受对腰果、核桃和花生的激发,随后监测过敏反应。通过酶联免疫吸附测定法(ELISA)对过敏抗体进行定量,并通过蛋白质印迹法鉴定蛋白质过敏原。通过脾细胞增殖试验和分泌细胞因子的测定来研究细胞反应。
单致敏小鼠在激发过程中对腰果和核桃有反应,而对花生激发产生的反应明显较弱。通过ELISA检测到对核桃和花生的交叉反应性IgE,并将交叉反应性过敏原鉴定为豌豆球蛋白。在细胞试验中,单致敏小鼠的脾细胞在接触腰果、核桃和花生后增殖并产生白细胞介素-4(IL-4)和白细胞介素-5(IL-5)。与单致敏小鼠相比,腰果加核桃致敏的小鼠对核桃有更强的临床反应,识别出更多的核桃过敏原,并且在核桃刺激的脾细胞试验中分泌的IL-4和IL-5明显更多。
发现腰果和核桃之间存在体内交叉反应性,而在T细胞水平上证明了腰果、核桃和花生之间存在交叉反应性。