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Constitutive delivery of bovine beta-lactoglobulin to the digestive tracts of gnotobiotic mice by engineered Lactobacillus casei

机译:工程化干酪乳杆菌将牛β-乳球蛋白组成性地递送至生生物小鼠的消化道

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摘要

The gut microbiota is critical for maturation of the immune system. Recent evidence suggests that early establishment of lactobacilli in the intestinal microbiota, during neonatal colonization or by probiotic supplementation, could prevent the development of allergic disorders. Postnatal maturation of the gut immune system with allergen-producing lactobacilli colonizing the digestive tract could then affect the development of further allergic sensitization. In this paper, we describe construction of a recombinant Lactobacillus casei strain that can constitutively deliver bovine beta-lactoglobulin (BLG), a major cow's milk allergen, to the guts of gnotobiotic mice. The blg gene was inserted into the L. casei chromosome downstream of an endogenous promoter. BLG production was improved by fusing the propeptide LEISSTCDA (LEISS) to the BLG mature moiety. This led to a 10-fold increase in LEISS-BLG production compared to the production obtained without the propeptide and also led to enhanced secretion corresponding to 5% of the total production. After inoculation into germfree C3H/HeN mice, the genetic stability of the recombinant strain and in vivo BLG production were confirmed for at least 10 weeks. BLG stimulation of spleen cells from mice monoassociated with the BLG-producing lactobacilli induced secretion of the Th1 cytokine gamma interferon and, to a lesser extent, the Th2 cytokine interleukin-5. No BLG-specific immunoglobulin G1 (IgG1), IgG2a, or IgA was detected in sera or in fecal samples. These results suggest that gut colonization with allergen-producing lactobacilli could provide a useful model for studying the modulation of allergic disorders.
机译:肠道菌群对于免疫系统的成熟至关重要。最近的证据表明,在新生儿定植期间或通过补充益生菌,在肠道菌群中早期建立乳酸杆菌可以预防过敏性疾病的发展。产后消化道定殖的产生变应原的乳酸菌对肠道免疫系统的产后成熟可能会影响进一步变态反应致敏的发展。在本文中,我们描述了一种重组干酪乳杆菌菌株的构建,该菌株可组成型地将牛β-乳球蛋白(BLG)(一种主要的牛奶过敏原)输送至生理性小鼠的肠道。将blg基因插入内源启动子下游的干酪乳杆菌染色体中。通过将前肽LEISSTCDA(LEISS)融合到BLG成熟部分来改善BLG的生产。与不使用前肽获得的产量相比,这导致LEISS-BLG产量增加了10倍,并且导致分泌增加,相当于总产量的5%。接种到无菌C3H / HeN小鼠中后,确认重组菌株的遗传稳定性和体内BLG产生至少10周。来自小鼠的脾脏细胞的BLG刺激与产生BLG的乳杆菌单联会诱导Th1细胞因子γ干扰素的分泌,并在较小程度上诱导Th2细胞因子白细胞介素5的分泌。在血清或粪便样本中未检测到BLG特异性免疫球蛋白G1(IgG1),IgG2a或IgA。这些结果表明,用产生变应原的乳杆菌对肠道进行定殖可以为研究变应性疾病的调节提供有用的模型。

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