虾制品致敏性诱发豚鼠相关细胞因子及Th1/Th2细胞平衡的变化
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摘要
以消减致敏性南美白对虾的虾仁、虾肉和虾蛋白为样品,以pH 7.5的磷酸盐缓冲液为阴性对照,未处理的虾蛋白为阳性对照,建立豚鼠过敏模型,研究过敏豚鼠血清中相关细胞因子与食物过敏的相关性,并推断食物过敏对辅助性T(type 1/type 2 T-helper, Th1/Th2)细胞平衡的影响。收集过敏豚鼠的血清,采用酶联免疫吸附测定法检测血清中免疫球蛋白E(immunoglobulin E,IgE)、组胺(histamine,HIS)、相关细胞因子(白细胞介素(interleukin,IL)-1、IL-2、IL-3、IL-4、IL-6、IL-10、肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)、干扰素-γ(interferon-γ,IFN-γ)质量浓度。结果表明,用未处理的虾蛋白致敏豚鼠,用致敏性消减程度不同的虾制品(虾蛋白、虾肉、虾仁)提取的蛋白激发,消减致敏性的虾蛋白、虾肉和虾仁提取蛋白激发后豚鼠血清中IgE含量分别为(3.905±0.120)、(4.813±0.188)、(5.199±0.327)U/mL,HIS质量浓度分别为(16.437±1.120)、(19.656±1.080)、(21.071±1.732)μg/mL,激发后血清中IgE和HIS质量浓度变化与致敏性程度呈正相关,致敏性越低,血清中IgE和HIS质量浓度越低。过敏豚鼠血清中IL-1、IL-2、IL-3、IL-4、IL-6、TNF-α质量浓度的变化与虾制品致敏程度呈正相关;同时,与IgE和HIS质量浓度变化规律具有一致性;血清中IL-10质量浓度变化与致敏性程度呈负相关;因此,这些细胞因子与食物过敏具有很好的相关性。过敏血清中IFN-γ没有呈现规律性变化,但IFN-γ/IL-4随着致敏性的增强而减小。因此,推测虾制品激发过敏豚鼠的Th1/Th2细胞平衡向Th2细胞偏移。
The objective of this work is to explore the correlation of serum cytokines in guinea pigs with food allergy and to deduce the impact of food allergy on the type 1/type 2 T-helper(Th1/Th2) cell balance. Mouse models of food allergy were induced by Pacific white shrimp(Litopenaeus vannamei) products with reduced allergenicity, shelled shrimp, shrimp meat and shrimp protein. Phosphate buffer(pH 7.5) was used as the negative control, and untreated shrimp protein was used as the positive control. Serum was collected from the allergic guinea pigs for the detection of immunoglobulin E(IgE), histamine(HIS) and cytokines(interleukin(IL)-1, IL-2, IL-3, IL-4, IL-6, IL-10, tumor necrosis factor α(TNF-α) and interferon-γ(IFN-γ)) by enzyme-linked immunosorbent assay(ELISA). Results showed that serum IgE levels in the guinea pigs sensitized by untreated shrimp protein and stimulated by shrimp protein with reduced allergenicity and the proteins extracted from shrimp meat and shelled shrimp with reduced allergenicity were(3.905 ± 0.120),(4.813 ± 0.188) and(5.199 ± 0.327) U/m L and HIS levels were(16.437 ± 1.120),(19.656 ± 1.080) and(21.071 ± 1.732) μg/mL, respectively. There was a positive correlation between the changes in serum IgE and HIS and allergenicity in the challenged animals. The changes in the serum levels of IL-1, IL-2, IL-3, IL-4, IL-6 and TNF-α were positively correlated with the allergenicity of shrimp products. These changes in cytokines were consistent with the changes in IgE and HIS contents. On the other hand, the change in IL-10 was negatively correlated with the allergenicity of shrimp products. Therefore, these seven cytokines had a good correlation with food allergy. Serum IFN-γ level did not show any correlation with the allergenicity of shrimp products, while the ratio of IFN-γ to IL-4 was decreased with the increase of allergenicity. Therefore, it is speculated that the Th1/Th2 cell balance in guinea pigs with food allergy is shifted towards Th2 cells.
The objective of this work is to explore the correlation of serum cytokines in guinea pigs with food allergy and to deduce the impact of food allergy on the type 1/type 2 T-helper(Th1/Th2) cell balance. Mouse models of food allergy were induced by Pacific white shrimp(Litopenaeus vannamei) products with reduced allergenicity, shelled shrimp, shrimp meat and shrimp protein. Phosphate buffer(pH 7.5) was used as the negative control, and untreated shrimp protein was used as the positive control. Serum was collected from the allergic guinea pigs for the detection of immunoglobulin E(IgE), histamine(HIS) and cytokines(interleukin(IL)-1, IL-2, IL-3, IL-4, IL-6, IL-10, tumor necrosis factor α(TNF-α) and interferon-γ(IFN-γ)) by enzyme-linked immunosorbent assay(ELISA). Results showed that serum IgE levels in the guinea pigs sensitized by untreated shrimp protein and stimulated by shrimp protein with reduced allergenicity and the proteins extracted from shrimp meat and shelled shrimp with reduced allergenicity were(3.905 ± 0.120),(4.813 ± 0.188) and(5.199 ± 0.327) U/m L and HIS levels were(16.437 ± 1.120),(19.656 ± 1.080) and(21.071 ± 1.732) μg/mL, respectively. There was a positive correlation between the changes in serum IgE and HIS and allergenicity in the challenged animals. The changes in the serum levels of IL-1, IL-2, IL-3, IL-4, IL-6 and TNF-α were positively correlated with the allergenicity of shrimp products. These changes in cytokines were consistent with the changes in IgE and HIS contents. On the other hand, the change in IL-10 was negatively correlated with the allergenicity of shrimp products. Therefore, these seven cytokines had a good correlation with food allergy. Serum IFN-γ level did not show any correlation with the allergenicity of shrimp products, while the ratio of IFN-γ to IL-4 was decreased with the increase of allergenicity. Therefore, it is speculated that the Th1/Th2 cell balance in guinea pigs with food allergy is shifted towards Th2 cells.
引文
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