Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Mycobacteriology and Pulmonary Research, Microbiology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Probiotics Antimicrob Proteins. 2024 Apr;16(2):367-382. doi: 10.1007/s12602-023-10058-6. Epub 2023 Mar 8.
Probiotics play a crucial role in immunomodulation by regulating dendritic cell (DC) maturation and inducing tolerogenic DCs. Akkermansia muciniphila affects inflammatory response by elevating inhibitory cytokines. We aimed to evaluate whether Akkermansia muciniphila and its outer membrane vesicles (OMVs) affect microRNA-155, microRNA-146a, microRNA-34a, and let-7i expression of inflammatory and anti-inflammatory pathways. Peripheral blood mononuclear cells (PBMCs) were isolated from the healthy volunteers. To produce DCs, monocytes were cultivated with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). DCs were allocated into six subgroups: DC + Lipopolysaccharide (LPS), DC + dexamethasone, DC + A. muciniphila (MOI 100, 50), DC + OMVs (50 µg/ml), and DC + PBS. The surface expression of human leukocyte antigen-antigen D related (HLA-DR), CD86, CD80, CD83, CD11c, and CD14 was examined using flow cytometry, and the expression of microRNAs was assessed using qRT-PCR, and the levels of IL-12 and IL-10 were measured using ELISA. A. muciniphila (MOIs 50, 100) could significantly decrease IL-12 levels relative to the LPS group. The IL-10 levels were decreased in the DC + LPS group than the DC + dexamethasone group. Treatment with A. muciniphila (MOI 100) and OMVs could elevate the concentrations of IL-10. DC treatment with LPS led to a significant increment in the expression of microRNA-155, microRNA-34a, and microRNA-146a. The expression of these microRNAs was reversed by A. muciniphilia and its OMVs treatment. Let-7i increased in treatment groups compared to the DC + LPS group. A. muciniphilia (MOI 50) had a substantial effect on the expression of HLA-DR, CD80, and CD83 on DCs. Therefore, DCs treatment with A. muciniphila led to induce tolerogenic DCs and the production of anti-inflammatory IL-10.
益生菌通过调节树突状细胞 (DC) 的成熟和诱导耐受性 DC 来发挥免疫调节作用。 Akkermansia muciniphila 通过提高抑制性细胞因子来影响炎症反应。我们旨在评估 Akkermansia muciniphila 及其外膜囊泡 (OMV) 是否影响炎症和抗炎途径的 microRNA-155、microRNA-146a、microRNA-34a 和 let-7i 的表达。从健康志愿者中分离外周血单核细胞 (PBMC)。为了产生 DC，单核细胞用粒细胞-巨噬细胞集落刺激因子 (GM-CSF) 和白细胞介素 4 (IL-4) 培养。将 DC 分为六组：DC+脂多糖 (LPS)、DC+地塞米松、DC+A. muciniphila (MOI 100、50)、DC+OMV (50µg/ml) 和 DC+PBS。使用流式细胞术检测人白细胞抗原-D 相关抗原 (HLA-DR)、CD86、CD80、CD83、CD11c 和 CD14 的表面表达，使用 qRT-PCR 评估 microRNA 的表达，并使用 ELISA 测量 IL-12 和 IL-10 的水平。与 LPS 组相比，A. muciniphila (MOIs 50、100) 可显著降低 IL-12 水平。与 DC+地塞米松组相比，DC+LPS 组的 IL-10 水平降低。用 A. muciniphila (MOI 100) 和 OMV 处理可提高 IL-10 的浓度。LPS 处理 DC 导致 microRNA-155、microRNA-34a 和 microRNA-146a 的表达显著增加。A. muciniphilia 和其 OMV 处理可逆转这些 microRNA 的表达。与 DC+LPS 组相比，治疗组中的 let-7i 增加。A. muciniphila (MOI 50) 对 DC 上 HLA-DR、CD80 和 CD83 的表达有显著影响。因此，A. muciniphila 处理 DC 可诱导耐受性 DC 并产生抗炎性 IL-10。
