Division of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
Clin Exp Allergy. 2022 Jan;52(1):137-148. doi: 10.1111/cea.13967. Epub 2021 Jul 1.
Two to four percentage of infants are affected by cow's milk allergy (CMA), which persists in 20% of cases. Intervention approaches using early oral exposure to cow's milk protein or hydrolysed cow's milk formula are being studied for CMA prevention. Yet, concerns regarding safety and/or efficacy remain to be tackled in particular for high-risk non-exclusively breastfed infants. Therefore, safe and effective strategies to improve early life oral tolerance induction may be considered.
We aim to investigate the efficacy of CMA prevention using oral pre-exposure of two selected 18-AA β-lactoglobulin-derived peptides loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) in a whey-protein induced CMA murine model.
The peptides were loaded in PLGA NPs via a double emulsion solvent evaporation technique. In vivo, 3-week-old female C3H/HeOuJ mice received 6 daily gavages with PBS, whey, Peptide-mix, a high- or low-dose Peptide-NPs or empty-NP plus Peptide-mix, prior to 5 weekly oral sensitizations with cholera toxin plus whey or PBS (sham). One week after the last sensitization, the challenge induced acute allergic skin response, anaphylactic shock score, allergen-specific serum immunoglobulins and ex vivo whey-stimulated cytokine release by splenocytes was measured.
Mice pre-treated with high-dose Peptide-NPs but not low-dose or empty-NP plus Peptide-mix, were protected from anaphylaxis and showed a significantly lower acute allergic skin response upon intradermal whey challenge compared to whey-sensitized mice. Compared with the Peptide-mix or empty-NP plus Peptide-mix pre-treatment, the high-dose Peptide-NPs-pre-treatment inhibited ex vivo whey-stimulated pro-inflammatory cytokine TNF-α release by splenocytes.
CONCLUSION & CLINICAL RELEVANCE: Oral pre-exposure of mice to two β-lactoglobulin-derived peptides loaded PLGA NPs induced a dose-related partial prevention of CMA symptoms upon challenge to whole whey protein and silenced whey-specific systemic immune response. These findings encourage further development of the concept of peptide-loaded PLGA NPs for CMA prevention towards clinical application.
有 2%至 4%的婴儿受到牛奶过敏（CMA）的影响，其中 20%的病例持续存在。目前正在研究通过早期口服接触牛奶蛋白或水解牛奶配方来干预 CMA 预防。然而，对于非纯母乳喂养的高风险婴儿，安全性和/或疗效方面的担忧仍有待解决。因此，可能需要考虑改善早期口服耐受诱导的安全有效的策略。
我们旨在通过口服预先暴露于两种选定的 18-AA β-乳球蛋白衍生肽负载聚（乳酸-共-乙醇酸）（PLGA）纳米颗粒（NPs），在乳清蛋白诱导的 CMA 小鼠模型中研究其预防 CMA 的效果。
通过双乳液溶剂蒸发技术将肽负载到 PLGA NPs 中。在体内，3 周龄雌性 C3H/HeOuJ 小鼠接受 6 次每日灌胃 PBS、乳清、肽混合物、高剂量或低剂量肽-NPs 或空-NP 加肽混合物，然后用霍乱毒素加乳清或 PBS（假对照）进行 5 周口服致敏。最后一次致敏后 1 周，通过测量过敏性皮肤反应、过敏休克评分、过敏原特异性血清免疫球蛋白和脾细胞中乳清刺激的细胞因子释放，评估过敏反应。
与乳清致敏小鼠相比，用高剂量肽-NPs 预处理而不是低剂量或空-NP 加肽混合物预处理的小鼠可免受过敏反应，并且在皮内乳清挑战时急性过敏皮肤反应明显降低。与肽混合物或空-NP 加肽混合物预处理相比，高剂量肽-NPs 预处理抑制了脾细胞中乳清刺激的促炎细胞因子 TNF-α的释放。
用负载有两种 β-乳球蛋白衍生肽的 PLGA NPs 对小鼠进行口服预暴露，可引起对全乳清蛋白的过敏症状呈剂量依赖性部分预防，并使乳清特异性全身免疫反应沉默。这些发现鼓励进一步开发负载肽的 PLGA NPs 用于 CMA 预防的概念，以推向临床应用。
