The HopQ-CEACAM Interaction Controls CagA Translocation, Phosphorylation, and Phagocytosis of Helicobacter pylori in Neutrophils
The HopQ-CEACAM Interaction Controls CagA Translocation, Phosphorylation, and Phagocytosis of Helicobacter pylori in Neutrophils
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HopQ-CEACAM相互作用控制中性粒细胞中幽门螺杆菌的CagA易位、磷酸化和吞噬作用
DOI：
10.1128/mbio.03256-19
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发表时间：
2020-01-01
期刊：
影响因子：
6.4
通讯作者：
Haas, Rainer
中科院分区：
文献类型：
--
作者：
Behrens, Ina-Kristin;Busch, Benjamin;Haas, Rainer
幽门螺杆菌的CagIV型分泌系统(CAG-T4SS)利用特定的细胞癌胚抗原相关细胞黏附分子(CEACAM)，如CEACAM1、-3、-5和-6，作为CagA转位到人胃上皮细胞的细胞受体。我们研究了幽门螺杆菌与人CEACAM1、CEACAM3和CEACAM6受体(HCEACAM)的相互作用。人和CEACAM人源化的小鼠多形核中性粒细胞(PMN)允许特定的依赖于HopQ的相互作用，强烈促进CagA易位。易位的CagA是酪氨酸磷酸化的，这在野生型(Wt)小鼠中性粒细胞中不存在。相比之下，人或小鼠骨髓来源的巨噬细胞和树突状细胞(DC)显示出低水平的hCEACAM表达和细菌结合。CAGA易位和酪氨酸-磷酸化水平较低，且不受HopQ-CEACAM相互作用的影响。CEACAM人源化小鼠的中性粒细胞，而不是巨噬细胞或DC，显著上调了促炎趋化因子MIP-1α。然而，在CEACAM人源化的巨噬细胞中，与wt细胞相比，巨噬细胞分泌的CXCL1(KC)和CCL2(MCP-1)显著减少。因此，幽门螺杆菌通过HopQ-CEACAM相互作用，以不同的方式控制中性粒细胞、巨噬细胞和DC中趋化因子的产生和分泌。我们进一步表明，与Hp接触后，中性粒细胞的氧化爆发和Hp的吞噬作用显著增强，但中性粒细胞上hCEACAM3/6的表达允许Hp在中性粒细胞内以HopQ依赖的方式延长存活时间。最后，我们证明在小鼠慢性感染期间，幽门螺杆菌能够系统地下调中性粒细胞上hCEACAM1和hCEACAM6受体的表达，可能限制CagA转位效率和很可能的胃病理学。然而，幽门螺杆菌的小鼠模型是针对幽门螺杆菌的人工模型，而且很少有适应菌株允许胃定植。在这里，我们证明了人或CEACAM人源化的中性粒细胞，而不是小鼠的中性粒细胞是由H.Pylori HopQ-CEACAM相互作用操纵的。人CEACAM负责中性粒细胞中CagA的磷酸化、激活和加工，而DC和巨噬细胞中的CagA转位和酪氨酸磷酸化不依赖于HopQ-CEACAM相互作用。幽门螺杆菌影响CEACAM人源化的中性粒细胞和巨噬细胞分泌不同的趋化因子。最重要的是，中性粒细胞上的人CEACAM增强了幽门螺杆菌的结合、氧化破裂和吞噬作用，并提高了细菌在吞噬小体中的存活率。幽门螺杆菌-CEACAM相互作用调节中性粒细胞，降低体内H.Pylori CagA的转位效率，并微调中性粒细胞上CEACAM受体的表达，以限制CagA的转位和胃病理。
The cag type IV secretion system (cag-T4SS) of Helicobacter pylori exploits specific cellular carcinoembryonic antigen-related cell adhesion molecules (CEACAMs), such as CEACAM1, -3, -5, and -6, as cellular receptors for CagA translocation into human gastric epithelial cells. We studied the interaction of H. pylori with human CEACAM1, CEACAM3, and CEACAM6 receptors (hCEACAMs) expressed on myeloid cells from CEACAM-humanized mice. Human and CEACAM-humanized mouse polymorphonuclear neutrophils (PMNs) allowed a specific HopQ-dependent interaction strongly enhancing CagA translocation. Translocated CagA was tyrosine phosphorylated, which was not seen in wild-type (wt) murine neutrophils. In contrast, human or murine bone marrow-derived macrophages and dendritic cells (DCs) revealed a low hCEACAM expression and bacterial binding. CagA translocation and tyrosine-phosphorylation was low and independent of the HopQ-CEACAM interaction. Neutrophils, but not macrophages or DCs, from CEACAM-humanized mice, significantly upregulated the proinflammatory chemokine MIP-1 alpha. However, macrophages showed a significantly reduced amount of CXCL1 (KC) and CCL2 (MCP-1) secretion in CEACAM-humanized versus wt cells. Thus, H. pylori, via the HopQ-CEACAM interaction, controls the production and secretion of chemokines differently in PMNs, macrophages, and DCs. We further show that upon H. pylori contact the oxidative burst of neutrophils and phagocytosis of H. pylori was strongly enhanced, but hCEACAM3/6 expression on neutrophils allowed the extended survival of H. pylori within neutrophils in a HopQ-dependent manner. Finally, we demonstrate that during a chronic mouse infection, H. pylori is able to systemically downregulate hCEACAM1 and hCEACAM6 receptor expression on neutrophils, probably to limit CagA translocation efficiency and most likely gastric pathology.IMPORTANCE Helicobacter pylori is highly adapted to humans and evades host immunity to allow its lifelong colonization. However, the H. pylori mouse model is artificial for H. pylori, and few adapted strains allow gastric colonization. Here, we show that human or CEACAM-humanized, but not mouse neutrophils are manipulated by the H. pylori HopQ-CEACAM interaction. Human CEACAMs are responsible for CagA phosphorylation, activation, and processing in neutrophils, whereas CagA translocation and tyrosine phosphorylation in DCs and macrophages is independent of the HopQ-CEACAM interaction. H. pylori affects the secretion of distinct chemokines in CEACAM-humanized neutrophils and macrophages. Most importantly, human CEACAMs on neutrophils enhance binding, oxidative burst, and phagocytosis of H. pylori and enhance bacterial survival in the phagosome. The H. pylori-CEACAM interaction modulates PMNs to reduce the H. pylori CagA translocation efficiency in vivo and to fine-tune the expression of CEACAM receptors on neutrophils to limit translocation of CagA and gastric pathology.