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ADP-heptose attenuates H. pylori-induced type I IFN signaling in DCs and impairs subsequent T cell responses

ABSTRACT: Helicobacter pylori (H. pylori) colonizes the gastric mucosa of approximately 50% of the world’s population. While it is well established that H. pylori is a major cause of gastric cancer, it has only recently been reported that H. pylori can also have adverse effects on immunological processes, including autoimmune diseases and immunotherapies. Understanding the paradoxical interplay between H. pylori and the human immune system will therefore be beneficial to improve a variety of therapeutic approaches. Here we show that ADP-heptose, a metabolite originally reported to act as a bona fide PAMP, attenuates full-fledged activation of primary human dendritic cells in the context of H. pylori infection by impairing type I IFN signaling, which in turn results in reduced DC maturation and T cell activation. Thus, this study provides novel mechanistic insights into how H. pylori utilizes ADP-heptose to mitigate host immunity.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Dendritic Cell, Blood

SUBMITTER: Veronika Schäpertöns

LAB HEAD: Dr. Christof Regl, MSc

PROVIDER: PXD046603 | Pride | 2024-09-13

REPOSITORIES: Pride

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			Action	DRS
	20201103_CR_H11M9_PepMap500_DCs_H_pyl_31.raw	Raw
	20201103_CR_H11M9_PepMap500_DCs_H_pyl_32.raw	Raw
	20201103_CR_H11M9_PepMap500_DCs_H_pyl_33.raw	Raw
	20201103_CR_H11M9_PepMap500_DCs_H_pyl_61.raw	Raw
	20201103_CR_H11M9_PepMap500_DCs_H_pyl_62.raw	Raw

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Project description:We previously established long-term 3D organoid culture systems for several murine tissues (intestine, stomach, pancreas and liver) as well as human intestine and pancreas. Here, we describe culture conditions to generate long-term 3D culture from human gastric stem cells. The technology can be applied to study the epithelial response to infection with Helicobacter pylori. Human gastric cultures can expand indefinitely in 3D Matrigel. Cultures can be generated from normal tissue, from single sorted stem cells, or from tumor tissue. Organoids maintain many characteristics of the respective tissue in terms of histology, marker expression and euploidy. Organoids from normal tissue express markers of four lineages of the stomach and self-organize in gland and pit-domains. They can be directed to specifically express either lineages of the gastric gland, or the gastric pit by addition of Nicotinamide and withdrawal of Wnt. While gastric pit lineages react marginally to bacterial infection, gastric gland lineages mount a strong inflammatory response. The gastric culture system provides a unique tool to study gastric pathologies. We generated 2 sets of experiments. The first set contains organoids in 4 conditions: (1) organoids in expansion condition ENRWFGNiTi ("gland-type organoids") from 3 donors, (2) organoids as in 1 but differentiated for 4 days in differentiation condition ENR_FGNiTi ("pit'type organoids"), (3) organoids as in 1 but infected with Helicobacter pylori strain P12 MOI 50 for 2 h, (4) organoids as in 2 but infected as in 3. All 4 conditions were tested on the same organoid line in parallel. This experiment was conducted independently with cultures from 3 different donors. The second set of experiments compares freshly isolated glands with organoids. Samples from 2 patients were analyzed. Each patient received a total gastrectomy. From each patient, glands from corpus region or pyloric antrum were isolated. From each isolation, one aliquod was stored for microarray analysis and one aliquod used to generate organoids. Organoids and glands were subsequently lysed and analyzed in parallel.

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ADP-heptose attenuates H. pylori-induced type I IFN signaling in DCs and impairs subsequent T cell responses

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