Department of Microbiology and Biotechnology, Max Rubner-Institut, Kiel, Germany.
Clinic of Dermatology, Venerology und Allergology, University Hospital Schleswig-Holstein, Kiel, Germany.
Yale J Biol Med. 2024 Sep 30;97(3):309-324. eCollection 2024 Sep.
Bacterial and food allergens are associated with immune-mediated food allergies via the gut-skin axis. However, there has been no data on the potential use of phages to rescue this pathological process. A human triple cell co-culture model incorporating colonocytes (T84 cells), macrophages (THP-1 cells), and hepatocytes (Huh7 cells) was established and infected with PAO1 (P.a PAO1) in the absence or presence of its KPP22 phage in Dulbecco's Modified Eagle's Medium (DMEM), DMEM+ ovalbumin (OVA), or DMEM+β-casein media. The physiological health of cells was verified by assessing cell viability and Transepithelial electrical resistance (TEER) across the T84 monolayer. The immune response of cells was investigated by determining the secretions of IL-1β, IL-8, IL-22, and IL-25. The ability of P.a PAO1 to adhere to and invade T84 cells was evaluated. The addition of either OVA or β-casein potentiated the P.a PAO1-elicited secretion of cytokines. The viability and TEER of the T84 monolayer were lower in the P.a PAO1+OVA group compared to the P.a PAO1 alone and PAO1+β-casein groups. OVA and β-casein significantly increased the adherence and invasion of P.a PAO1 to T84 cells. In the presence of the KPP22 phage, these disruptive effects were abolished. These results imply that: (1) food allergens and bacterial toxic effector molecules exacerbate each other's disruptive effects; (2) food allergen and bacterial signaling at the gut-skin mucosal surface axis depend on a network of bacteria-phage-eukaryotic host interactions; and (3) phages are complementary for the evaluation of pathobiological processes that occur at the interface between bacteria, host cellular milieu, and food antigens because phages intervene in P.a PAO1-, OVA-, and β-casein-derived inflammation.
细菌和食物过敏原通过肠-皮轴与免疫介导的食物过敏有关。然而，目前还没有关于噬菌体潜在用途的数据可以用来挽救这种病理过程。建立了一个包含结肠细胞（T84 细胞）、巨噬细胞（THP-1 细胞）和肝细胞（Huh7 细胞）的人类三重细胞共培养模型，并在无或存在其 KPP22 噬菌体的情况下在 Dulbecco 的改良 Eagle 培养基（DMEM）、DMEM+卵清蛋白（OVA）或 DMEM+β-酪蛋白培养基中感染 PAO1（P.a PAO1）。通过评估 T84 单层细胞跨上皮电阻（TEER）和细胞活力来验证细胞的生理健康。通过测定 IL-1β、IL-8、IL-22 和 IL-25 的分泌来研究细胞的免疫反应。评估了 P.a PAO1 对 T84 细胞的黏附和侵袭能力。添加 OVA 或 β-酪蛋白增强了 P.a PAO1 引发的细胞因子分泌。与 P.a PAO1 单独组和 PAO1+β-酪蛋白组相比，P.a PAO1+OVA 组 T84 单层细胞的活力和 TEER 较低。OVA 和 β-酪蛋白显著增加了 P.a PAO1 对 T84 细胞的黏附和侵袭。在 KPP22 噬菌体存在的情况下，这些破坏作用被消除。这些结果表明：（1）食物过敏原和细菌毒性效应分子相互加剧彼此的破坏作用；（2）食物过敏原和细菌信号在肠-皮黏膜表面轴上取决于细菌-噬菌体-真核宿主相互作用网络；（3）噬菌体在评估发生在细菌、宿主细胞环境和食物抗原之间界面的病理生物学过程中是互补的，因为噬菌体干预了 P.a PAO1、OVA 和 β-酪蛋白衍生的炎症。
