Dietary Vitamin D Supplementation Is Ineffective in Preventing Murine Cow’s Milk Allergy, Irrespective of the Presence of Nondigestible Oligosaccharides

Abstract

<b><i>Introduction:</i></b> Cow’s milk allergy (CMA) is one of the most common food allergies especially early in life. A mixture of nondigestible short-chain galacto-oligosaccharides, long-chain fructo-oligosaccharides, and pectin-derived acidic-oligosaccharides (GFA) may reduce allergy development and allergic symptoms in murine CMA. Recently, vitamin D (VitD) has been suggested to have beneficial effects in reducing allergy as well. <b><i>Objective:</i></b> In this study, the immune modulatory effect on allergy prevention using the combination of GFA and VitD was investigated. <b><i>Methods:</i></b> Female C3H/HeOuJ mice were fed a control or GFA-containing diet with depleted, standard (1,000 IU/kg), or supplemented (5,000 IU/kg) VitD content for 2 weeks before and during whey sensitization (<i>n</i> = 10–15). Mice were sensitized 5 times intragastrically with PBS as a control, whey as cow’s milk allergen, and/or cholera toxin as adjuvant on a weekly interval. One week after the last sensitization, mice were intradermally challenged in both ear pinnae and orally with whey, subsequently the acute allergic skin response and shock symptoms were measured. After 18 h, terminal blood samples, mesenteric lymph nodes, and spleens were collected. Whey-specific immunoglobulin (Ig) E and IgG1 levels were measured by means of ELISA. T cell subsets and dendritic cells (DCs) were studied using flow cytometry. <b><i>Results:</i></b> Additional VitD supplementation did not lower the allergic symptoms compared to the standard VitD diet. CMA mice fed the GFA diet supplemented with VitD (GFA VitD⁺) significantly decreased the acute allergic skin response of whey sensitized mice when compared to the CMA mice fed VitD (VitD⁺) group (<i>p</i> &#x3c; 0.05). The effect of GFA was not improved by extra VitD supplementation even though the CMA mice fed the GFA VitD⁺ diet had a significantly increased percentage of CD103⁺ DCs compared to the VitD⁺ group (<i>p</i> &#x3c; 0.05). The VitD-deprived mice showed a high percentage of severe shock and many reached the humane endpoint; therefore, these groups were not further analyzed. <b><i>Conclusions:</i></b> High-dose VitD supplementation in mice does not protect against CMA development in the presence or absence of GFA.