---
title: "Helicobacter pylori immune priming depends on type I interferon signaling"
id: "frontiers-in-immunology-12-helicobacter-pylori-induced-immune-priming-requires-type-i-interferon-signaling"
canonical_url: "https://medichelpline.com/clinical-feed/frontiers-in-immunology-12-helicobacter-pylori-induced-immune-priming-requires-type-i-interferon-signaling"
content_type: "clinical_feed_article"
specialty: "Infectious Disease"
source_name: "Frontiers in Immunology"
source_url: "https://www.frontiersin.org/articles/10.3389/fimmu.2026.1942802"
published_at: "2026-10-02T00:00:00.000Z"
evidence_level: "Journal Feed"
license: "CC-BY-NC-4.0 / Informational Use"
---
# Helicobacter pylori immune priming depends on type I interferon signaling
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/frontiers-in-immunology-12-helicobacter-pylori-induced-immune-priming-requires-type-i-interferon-signaling
- **Specialty:** [Infectious Disease](https://medichelpline.com/clinical-feed/infectious-disease.md)
- **Primary Source:** Frontiers in Immunology
- **Source URL:** [Original Journal Publication](https://www.frontiersin.org/articles/10.3389/fimmu.2026.1942802)
- **Published At:** 2026-10-02T00:00:00.000Z
- **Evidence Rating:** Journal Feed
## Executive GIST (TL;DR)
- The source is an original research article titled “Helicobacter pylori-induced immune priming requires type I interferon signaling” published in Frontiers in Immunology, as indicated by the provided page metadata and URL. - The article title states the central finding: **Helicobacter pylori**-induced **immune priming** requires **type I interferon** signaling. This is the only explicit scientific claim available in the source content provided. - The provided source material contains website navigation, journal sections, and links but does not include the article abstract, methods, results, figures, author names, institutions, or conclusions beyond the title. - The DOI-like path in the URL includes 10.3389/fimmu.2026.1942802, implying a Frontiers in Immunology article entry; no additional numeric results or dates are reported within the article body text supplied here. - Because the supplied content lacks study details, information on experimental design, sample sizes, models (human, animal, or in vitro), statistical analyses, effect sizes, or specific immunological mechanisms beyond the title claim was not reported and cannot be inferred. - Implications and context suggested by the title (for example, that **type I interferon signaling** is mechanistically important for H. pylori-driven changes in host immunity) are plausible but the source did not provide supporting data, methods, or discussion text to substantiate or contextualize those implications. - Any clinical or translational relevance, therapeutic considerations, or guideline impacts were not reported in the supplied source material and therefore cannot be stated here.
## Clinical Analysis & Structured Key Points
Frontiers | Helicobacter pylori-induced immune priming requires type I interferon signaling ORIGINAL RESEARCH article Front. Immunol. , 02 October 2026 Sec. Immunological Memory Volume 17 - 2026 | https://doi.org/10.3389/fimmu.2026.1942802 Published in Frontiers in Immunology Immunological Memory 7 impact factor 11.3 citescore Editor & Reviewers Edited by K K Karl Kai McKinstry Reviewed by C G Cuncai Guo J S Julia Sanchez-Garrido S G Somayeh Ghotloo Outline Figures and Tables Figure 1 View in article Figure 2 View in article Figure 3 View in article Figure 4 View in article Figure 5 View in article Table 1 List of primers. View in article ORIGINAL RESEARCH article Front. Immunol. , 02 October 2026 Sec. Immunological Memory Volume 17 - 2026 | https://doi.org/10.3389/fimmu.2026.1942802 Helicobacter pylori -induced immune priming requires type I interferon signaling Z R Zhigang Rao 1,2 C R Christof Regl 1,2 T N Theresa Neuper 1,2,3 H D Hieu-Hoa Dang 1,2 G U Gunda Ueblagger 1,2 A V Andreas Völkerer 3,4 C G Christian G. Huber 1,2 J H Jutta Horejs-Hoeck 1,2,3 * 1. Department of Biosciences and Medical Biology, Paris-Lodron University Salzburg, Salzburg, Austria 2. Center for Tumor Biology and Immunology (CTBI), Paris-Lodron University Salzburg, Salzburg, Austria 3. Life Sciences Center Salzburg (LSCS), Salzburg, Austria 4. General Hospital Oberndorf, Teaching Hospital of the Paracelsus Medical University, Salzburg, Austria See more Article metrics View details Abstract Helicobacter pylori ( H. pylori ) infection induces persistent colonization and chronic inflammation, yet the mechanisms by which bacterial exposure establishes sustained inflammatory responses remain incompletely understood. We previously demonstrated that priming human monocytes with viable, but not heat-killed H. pylori , establishes immune priming, characterized by a heightened response to subsequent LPS challenge. Here, we delineate the signaling pathways that establish this inflammatory adaptation. Unbiased whole-cell proteomics identified type I and type II interferon (IFN) signaling as the dominant pathways distinguishing monocytes exposed to viable versus heat-killed bacteria. Consistently, viable H. pylori , but not heat-killed or PFA-inactivated bacteria, activated IFN signaling and imprinted LPS hyper-responsiveness. Manipulating IFN signaling at the time of LPS challenge did not modify this hypersensitivity, whereas blocking type I IFN signaling during priming markedly attenuated the hyperresponsiveness. Accordingly, supplementation with type I, but not type II IFN, during priming restored hypersensitivity in monocytes exposed to PFA-inactivated H. pylori . Interestingly, type I IFN only partially restored LPS-hypersensitivity in monocytes exposed to heat-killed H. pylori , indicating that heat inactivation interferes with additional signaling cues derived from intact bacteria, which are required to drive the full hyper-responsiveness to LPS. Collectively, our findings identify type I IFN as a central regulator of H. pylori -induced immune priming and reveal a previously unrecognized mechanism by which H. pylori infection programs persistent inflammatory adaptation in host immune cells through type I IFN signaling. Introduction Although initially considered a hallmark of antiviral immunity, activation of type I interferon (type I IFN) signaling has been recognized as a common feature of many bacterial infections, in which type I IFNs are induced upon sensing of pathogen-associated molecular patterns (PAMP) by pattern-recognition receptors (PRR) ( 1 ). Type I IFNs bind to and activate the canonical Janus kinase (JAK)- signal transducers and activators of transcription (STAT) signaling, resulting in phosphorylation of STAT1 and STAT2 and formation of the interferon-stimulated gene factor 3 (ISGF3) complex with IRF9, thereby driving the transcription of interferon-stimulated genes (ISGs) ( 2 ). Given the ubiquitous expression of interferon-α/β receptor (IFNAR), type I IFN signaling exerts broad immunomodulatory effects ( 3 ), while type II interferon (type II IFN) signaling likewise serves as a major regulator of antimicrobial immunity and inflammatory responses ( 4 ). Despite signaling through distinct receptors and activating different STAT complexes, type I IFN and type II IFN signaling share substantial overlap in downstream gene expression and frequently cooperate to shape innate immune functions ( 5 ). While certain intracellular pathogens exploit type I IFN signaling to promote their survival ( 6 – 8 ), accumulating evidence identifies its prominent roles in immunomodulatory ( 9 ) and immune priming and memory ( 10 ). For instance, type I IFN-dependent reprogramming has been implicated in the regulation of hematopoietic stem and progenitor cell responses during Mycobacterium tuberculosis infection ( 11 ), in β-glucan-induced hematopoietic expansion ( 12 ), and in shaping the magnitude of peripheral blood mononuclear cells (PBMC) or macrophage responses to secondary inflammatory stimulation ( 13 – 16 ). These observations indicate that type I IFN signaling can induce long-lasting functional adaption of innate immune cells beyond its established role in antimicrobial defense. Therefore, it is plausible to speculate that type I IFN contributes to inflammatory adaptation during chronic bacterial infections, such as Helicobacter pylori ( H. pylori ), the infection of which usually establishes life-long colonization of the gastric mucosa and drives persistent inflammation that contributes to chronic disease progression ( 17 ). Accordingly, our previous work demonstrated that H. pylori priming induces immune priming in human monocytes, characterized by exaggerated TNF-α production following subsequent lipopolysaccharide (LPS) challenge ( 18 , 19 ). Unlike many gram-negative pathogens, H. pylori LPS is structurally modified and exhibits low or absent toll-like receptor (TLR)4 agonist activity, allowing the bacterium to evade robust TLR4-driven inflammation ( 20 ). Instead, H. pylori engages alternative sensing pathways, including TLR2 during bacteria uptake ( 21 ) and TLR8 for bacterial RNA ( 22 ). Indeed, H. pylori has been shown to activate type I IFN responses in monocytes through TLR8 ( 22 ), highlighting interferon signaling as an integral component of the host response to H. pylori infection. Although H. pylori -induced type I IFN responses and immune priming have each been described independently, whether these two phenomena are mechanistically linked remains unknown. In the present study, we show that type I IFN, but not type II IFN signaling during the priming phase with H. pylori is required for establishing enhanced responsiveness of monocytes to subsequent LPS challenge. These findings identify type I IFN signaling as a previously unrecognized regulator of H. pylori -induced immune priming and provide mechanistic insights into how persistent H. pylori infection may promote chronic inflammatory responses. Materials and methods Isolation of PBMC and monocytes Human PBMC were isolated from fresh buffy coats prepared from freshly collected peripheral blood of healthy anonymized volunteers, provided by the Blood Bank Salzburg. Briefly, PBMC were isolated via density gradient centrifugation (836 × g, 20 min, RT) using Histopaque-1077 (Sigma-Aldrich, Vienna, Austria, #10771). Monocytes were subsequently purified using CD14 MicroBeads (Miltenyi Biotec, Bergisch-Gladbach, Germany, #130-050-201) following the manufacturer´s instructions, with the exception that only 30% of the recommended amount of MicroBeads were used. The purity of CD14 + cells was > 98% and the viability was > 99%, as determined by flow cytometry. Cells were cultured in monocyte medium: RPMI-1640 medium (Sigma-Aldrich, #R0083) supplemented with 10% heat-inactivated fetal calf serum (Biowest, Nuaillé, France, #S1400) and 1% L-glutamine (Sigma-Aldrich, #G7513). Following isolation, monocytes were allowed to adhere for 1–2 h at 37 °C in a humidified atmosphere containing 5% CO 2 before further experimental use. Bacterial culture and infection H. pylori P12 wild type were cultivated under microaerophilic conditions using an Oxoid 3.5 L bacterial culture bucket (Thermo Fisher Scientific, Vienna, Austria, #AB0035A) containing Oxoid CampyGen 3.5 L sachets (Thermo Fisher Scientific, #CN0035A). Bacteria were grown on GC agar-plates supplemented with 10% horse serum (Biowest) at 37 °C. Frozen bacteria stocks were first cultured for 72 h, followed by subculturing onto fresh agar plates for another 24 h prior to use in monocyte infection experiments. Bacterial biomass was harvested from agar plates using sterile cotton buds (Paul Boettger, Bodenmais, Germany, #09129-0100) and suspended in 1 mL PBS (Sigma-Aldrich, #D8537). Optical density was determined at OD600 using a BioPhotometer Plus device (Eppendorf, Vienna, Austria, #6132) and bacteria concentrations were estimated based on an in-house OD600-to-cell number calibration curve. Unless otherwise stated, infection experiments were performed at a multiplicity of infection of 5. Heat-killed H. pylori ( Hp heat ) was generated by incubating bacterial suspensions in PBS at 95 °C for 5 min, followed by equilibration at RT for 3 min before use. Paraformaldehyde (PFA)-inactivated H. pylori ( Hp PFA ) was prepared by treating viable H. pylori suspensions with 4% paraformaldehyde (Thermo Fisher Scientific, #I28800) at RT for 15 min. Subsequently, bacteria were pelleted by centrifugation (10, 000 × g, RT, 5 min), washed twice with PBS, and resuspended in PBS before further use. For the generation of antibiotic-treated H. pylori ( Hp ab ), H. pylori was cultured in BHI medium (37.75 g/L) containing β-cyclodextrin (10 g/L; Thermo Fisher Scientific, #J63161), thrimetoprim (2.5 mg/L; pre-dissolved in DMF; Thermo Fisher Scientific, #T7883), vancomycin (10 mg/L; pre-dissolved in water; Roth, Karlsruhe, Germany, #0242.3), nystatin (1 mg/L; pre-dissolved in DMSO; Roth, #2411) and vitamin mix consisting of dextrose (1 g/L, #D-9434), L-glutamine (0.1 g/L, #G-3126), carboxylase (1 mg/L, #C8754), iron(III) nitrate nonahydrate (0.2 mg/L, #216828), thiamin hydrochloride (0.03 mg/L, #T-4625), 4-aminobenzoic acid sodium salt (0.13 mg/L, #A6928), β-nicotinamide adenine dinucleotide hydrate (2.5 mg/L, #N-7004), vitamin B12 (0.1 mg/L, #V-2876), L-cystine (11 mg/L, #C-6727), Adenine (10 mg/L, #A9795), Guanine (0.3 mg/L, #51030), Arginin (1.5 mg/L, A-6969), Uracil (5 mg/L, #U1128) (all from Sigma-Aldrich) and cysteine-HCl (0.26 g/L, Roth, #1694.2). Viable H. pylori (OD600 = 0.5 – 1) cultures were exposed to amoxicillin (0.05 mg/mL; pre-dissolved in DMSO; MedChemExpress, Monmouth Junction, NJ, #HY-B0467A) and clarithromycin (0.05 mg/mL; pre-dissolved in DMSO; MedChemExpress, #HY-17508) for 2 h. Following treatment, bacteria were collected by centrifugation (10, 000 × g, 5 min, RT), washed twice with PBS, and resuspended in PBS before use. To verify the loss of cultivability, Hp heat , Hp PFA and Hp ab preparations were replated onto GC agar-plates supplemented with 10% horse serum and incubated under microaerophilic conditions at 37 °C. No colony formation was detected within 72 h of incubation. Monocyte short-term immune priming model The human monocyte short-term immune priming model was established as previously described ( 18 , 19 ). In brief, monocytes were seeded at a density of 1 × 10 6 /mL (100 µL/well) in 96-well V-bottom cell culture plates (Greiner Bio-One, Kremsmünster, Austria, #651180) and rested for 1 h. Afterwards, cells were either left unprimed, or exposed to indicated H. pylori conditions at a multiplicity of infection of 5 for 24 h. Following the priming phase, plates were centrifugated (716 × g, RT, 5 min), and supernatants (200 µL) were collected for measurement of cytokines. Cells were then gently washed with 100 µL of challenge medium: RPMI-1640 medium (Sigma-Aldrich, #R0083) supplemented with 10% heat-inactivated fetal calf serum (Biowest, #S1400) and 1% L-glutamine (Sigma-Aldrich, #G7513) and 1% penicilin/streptomycin (Sigma-Aldrich, #P4333). Thereafter, cells were incubated in 200 µL challenge medium in the absence or presence of 10 ng/mL E. coli LPS (O55:B5, Sigma-Aldrich, #L2880) for another 24 h. At the end of the experiment, supernatants were collected for measurement of cytokines. To access the viability of H. pylori after priming and challenge, supernatants were collected 1) after priming and 2) after the subsequent challenging phase, and plated on GC agar plates supplemented with 10% horse serum under microaerophilic conditions at 37 °C for 6 days. Colony formation was monitored by photography. For pharmacological and cytokine intervention experiments, celecoxib (1 – 10 µM; MedChemExpress, #HY-14398), PGE 2 (1–10 nM; MedChemExpress, #HY-101952), IFN-α (10 ng/mL; Immuno Tools, Friesoythe, Germany, #11343504), IFN-β (10 ng/mL; Immuno Tools, #11343594), anifrolumab (20 ng/mL; MedChemExpress, #HY-P99168), IFN-γ (20 ng/mL; Immuno Tools, #11343534) were added either 15 min before the onset of the priming phase or shortly before the challenge phase, as indicated in the respective experiment. Determination of cytokine levels Cell culture supernatants were collected following monocyte stimulation and stored at -80 °C before analysis. Concentrations of tumor necrosis factor (TNF)-α and interferon gamma-induced protein 10 (IP-10) were quantified using commercially available Enzyme-linked Immunosorbent Assay kits (TNF-α: #900-K25K; IP-10: #900-K39; Peprotech, London, United Kingdom) according to the manufacturer´s instructions. Whole-cell proteomic analysis and bioinformatics Monocytes were infected with the corresponding Hp conditions for 24 h. Subsequently, cell pellets (1.5 × 10 6 ) were harvested and processed as previously described ( 21 ). Briefly, pellets were resuspended in 50 mM triethylammonium bicarbonate buffer (pH 7.55, Sigma-Aldrich, #T7408) containing 5% SDS (Sigma-Aldrich, #L3771) and 1 × cOmplete Mini EDTA-free protease inhibitor cocktail (Roche, #04693159001). Samples were then heated at 95 °C for 5 min, followed by sonication for 5 min using a Bioruptor device (Diagenode, Liège, Belgium). After centrifugation (14, 000 × g, 4 °C, 1 min), supernatants were harvested and protein concentrations were determined using the Pierce BCA Protein assay kit (Thermo Fisher Scientific, #23255) according to the manufacturer’s instructions. Protein lysates were then treated with tris-(2-carboxyethyl)-phosphine-hydrochloride (5 mM, Sigma-Aldrich, #75259) at 55 °C for 15 min to reduce disulfides, followed by alkylation of the cysteine residues with iodoacetamide (Sigma-Aldrich, #I6125) at a final concentration of 20 mM for 10 min at 22 °C in the dark. Samples were then acidified to pH ≤ 1 using 12% (v/v) orthophosphoric acid (Merck, Darmstadt, Germany, #100573), and proteins were precipitated by addition of 100 mM triethylammonium bicarbonate buffer (pH 7.5, in 90% methanol) (Merck, #106009) at a ratio of 7:1 (v/v). Protein purification was then performed by suspension trapping using S-Trap mini columns (Protifi, Huntington, NY, USA) according to the manufacturer’s instructions. Proteins were digested to peptides using sequencing grade modified porcine trypsin (Promega, Madison, WI, USA, #V5111) at a protease/protein ratio of 1:10 (w/w) at 37 °C for 16 h. Resulting peptides were dried in a vacuum centrifuge at 60 °C and resuspended in an aqueous solution containing 1% acetonitrile (VWR International, Vienna, Austria, #20060.320) and 0.1% formic acid (Sigma-Aldrich, #1002631000) (v/v) to a final concentration of 3.0 µg/µL. Chromatographic separation of 6.0 µg of peptides was performed by a reversed-phase nanoHPLC using a Vanquish Neo UHPLC System (Thermo Fisher Scientific), equipped with an Aurora Ultimate XT C18 column (250 × 0.075 mm i.d.) (ionopticks, #AUR3-25075C18-XT). Separation was conducted at a flow rate of 300 nL/min using 0.1% aqueous formic acid (solvent A) and 0.1% formic acid in acetonitrile (solvent B) with the following gradient: 1% B for 5 min, followed by a linear gradient of 1-35% B till 100 min. This was followed by a washing step with 80% B for 5 min and column re-equilibration at 1% B for 15 min. Column temperature was maintained at 50 °C, while the autosampler was kept at 7 °C. The nanoHPLC system was coupled to a Q Exactive™ Hybrid Quadrupole-Orbitrap™ mass spectrometer (Thermo Fisher Scientific) via a Nanospray Flex™ ion source (Thermo Fisher Scientific). Spray voltage was set to 1.5 kV, the S-lens RF level to 60.0 and the capillary temperature to 250 °C. Each acquisition cycle consisted of one full MS scan over a m/z range of 350–1, 200 at a resolution of 35, 000 at m/z 200, followed by 36 data-independent higher-energy collisional dissociation scans in 24.0 m/z isolation windows from m/z 350–1200 with 29% normalized collision energy and a resolution of 17, 500 at m/z 200. For all scans, the automatic gain control target was set to 1e6 charges with a maximum injection time of 60 ms. Data acquisition was performed using Thermo Scientific™ Chromeleon™ 7.2 CDS (Thermo Fisher Scientific). For data analysis, DIA-NN 2.2.0 ( 23 ) was used with the protease settings “Trypsin/P” with a maximum of three missed cleavages, carbamidomethylation of cysteine as fixed and oxidation of methionine as variable modification. Protein identification was performed against an in-silico generated spectral library of the reviewed Swiss-Prot UniProt consortium database ( 24 ) containing Homo sapiens (Human) entries downloaded on 22.07.2025, with a false discovery rate of 1%. The resulting protein groups were further processed in RStudio 2525.05.1 + 513.pro3 ( 25 ). First, the intensities were log2-transformed, followed by the removal of proteins that were not detected across at least 10% of all samples, and potential contaminants. Next, intensities were normalized by subtraction of the median intensity of each sample, followed by a principal component analysis. As principal component analysis revealed clustering associated not only with the treatment but also with donor availability, differential expression analysis was performed with limma in R using the donor as a blocking factor for paired analysis ( 26 ). The p -values were adjusted with the Benjamini-Hochberg procedure. Fast gene set enrichment analysis was performed using fgsea package ( 27 ) in R using the Reactome pathway database ( https://reactome.org/cite ). The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium ( http://proteomecentral.proteomexchange.org ) via the PRIDE partner repository ( 28 ) with the dataset identifier PXD080861. Sample preparation and analysis of PGE 2 Monocytes (1 × 10 6 in 0.5 mL culture medium) were pre-treated without or with 10 µM celecoxib for 15 min, and then infected without or with viable H. pylori , or Hp heat for 24 h. Cell supernatants (0.4 mL) were harvested in 0.5 mL of ice-cold methanol containing 800 pg of d 4 -PGE 2 (Fisher Scientific, 30427024) as an internal standard to facilitate metabolite quantitation and sample recovery. Solid phase extraction was conducted using adapted protocols as previously described ( 29 ). Briefly, samples were first kept at -20 °C for at least 2 h to allow protein sedimentation, followed by centrifugation (400 × g, 4 °C, 10 min). Supernatants were transferred to new tubes and mixed with 2 mL of acidified water (pH = 3.5) before loading onto C18 AR column cartridges (15 mg, Agilent Technology) pre-equilibrated with 1 mL methanol and 0.5 mL water. After sequential washing steps with 1 mL water and 1 mL hexane, metabolites were eluted with 1 mL methanol and brought to dryness using an Eppendorf concentrator (Thermo Fisher Scientific) using the program V-HV at 30 °C for 60–90 min. Dried lipid films were stored at -20 °C before dissolving in 100 µL methanol/water (50/50, v/
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