Comprehensive immune profiling and predictive modelling of paediatric acute hepatitis of unknown aetiology from a Spanish cohort

IntroductionPaediatric acute hepatitis of unknown aetiology (PAHUA) has emerged as a global health concern, yet its cause remains unidentified. This study characterises the clinical and immunological profiles of PAHUA to identify reliable immune biomarkers for accurate diagnosis.MethodsSamples from...

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Main Authors: Roberto Lozano-Rodríguez, Loreto Hierro, María José Quiles, Alejandro Pascual-Iglesias, Verónica Terrón-Arcos, Gema Muñoz-Bartolo, Esteban Frauca, Francisco J. Cueto, Cristina Calvo, Laura Córdoba-García, Jesús Fernández-Felipe, Laura Hurtado-Navarro, Julia del Prado-Montero, Gonzalo Sáenz de Santa María-Diez, Daniel Arvelo-Rosario, Paloma Jara, Carlos del Fresno, Eduardo López-Collazo
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1599982/full
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author Roberto Lozano-Rodríguez
Roberto Lozano-Rodríguez
Loreto Hierro
María José Quiles
Alejandro Pascual-Iglesias
Alejandro Pascual-Iglesias
Verónica Terrón-Arcos
Verónica Terrón-Arcos
Gema Muñoz-Bartolo
Esteban Frauca
Francisco J. Cueto
Francisco J. Cueto
Cristina Calvo
Cristina Calvo
Laura Córdoba-García
Laura Córdoba-García
Jesús Fernández-Felipe
Jesús Fernández-Felipe
Laura Hurtado-Navarro
Laura Hurtado-Navarro
Julia del Prado-Montero
Julia del Prado-Montero
Gonzalo Sáenz de Santa María-Diez
Gonzalo Sáenz de Santa María-Diez
Daniel Arvelo-Rosario
Daniel Arvelo-Rosario
Paloma Jara
Carlos del Fresno
Eduardo López-Collazo
Eduardo López-Collazo
Eduardo López-Collazo
Eduardo López-Collazo
Eduardo López-Collazo
author_facet Roberto Lozano-Rodríguez
Roberto Lozano-Rodríguez
Loreto Hierro
María José Quiles
Alejandro Pascual-Iglesias
Alejandro Pascual-Iglesias
Verónica Terrón-Arcos
Verónica Terrón-Arcos
Gema Muñoz-Bartolo
Esteban Frauca
Francisco J. Cueto
Francisco J. Cueto
Cristina Calvo
Cristina Calvo
Laura Córdoba-García
Laura Córdoba-García
Jesús Fernández-Felipe
Jesús Fernández-Felipe
Laura Hurtado-Navarro
Laura Hurtado-Navarro
Julia del Prado-Montero
Julia del Prado-Montero
Gonzalo Sáenz de Santa María-Diez
Gonzalo Sáenz de Santa María-Diez
Daniel Arvelo-Rosario
Daniel Arvelo-Rosario
Paloma Jara
Carlos del Fresno
Eduardo López-Collazo
Eduardo López-Collazo
Eduardo López-Collazo
Eduardo López-Collazo
Eduardo López-Collazo
author_sort Roberto Lozano-Rodríguez
collection DOAJ
description IntroductionPaediatric acute hepatitis of unknown aetiology (PAHUA) has emerged as a global health concern, yet its cause remains unidentified. This study characterises the clinical and immunological profiles of PAHUA to identify reliable immune biomarkers for accurate diagnosis.MethodsSamples from 24 PAHUA patients, 6 children with autoimmune hepatitis (AIH), and 13 healthy paediatric volunteers (HVs) were analysed. Immunophenotyping, soluble immune checkpoints (ICs) and cytokine profiling, and ex vivo immune responses were assessed using spectral flow cytometry. Clustering and logistic regression modelling were used to identify predictive biomarkers.ResultsPAHUA cases frequently presented with gastrointestinal symptoms and liver damage preceding jaundice, with 59% progressing to paediatric acute liver failure (pALF). Adenovirus was detected in only 17.6% of PAHUA patients, suggesting it is unlikely to be the primary causative agent. Antibodies against the SARS-CoV-2 Spike protein were identified in 88.2% of PAHUA patients, as well as in AIH and HV groups, indicating prior exposure. Immunophenotyping, ICs and cytokine profiling, and ex vivo immune revealed distinct immune profiles between PAHUA and non-PAHUA individuals. Furthermore, clustering and logistic regression modelling identified potential predictive biomarkers, including the plasmatic ICs Gal-9 and sTim-3, alongside specific immune cell populations. Notably, a combined Gal-9 and sTim-3 model achieved an AUC of 1.000 in differentiating PAHUA patients from non-PAHUA individuals.ConclusionsDespite the limited cohort analysed, owing to the rarity of cases worldwide, our data provide valuable insights for an accurate, early, and minimally invasive diagnosis of PAHUA. These patients exhibit a distinct immunological profile, with ICs, particularly Gal-9 and sTim-3, showing strong potential as reliable biomarkers.
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spelling doaj-art-551b7aebc94540e2961fc552a4ce2c7e2025-08-20T03:23:22ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-06-011610.3389/fimmu.2025.15999821599982Comprehensive immune profiling and predictive modelling of paediatric acute hepatitis of unknown aetiology from a Spanish cohortRoberto Lozano-Rodríguez0Roberto Lozano-Rodríguez1Loreto Hierro2María José Quiles3Alejandro Pascual-Iglesias4Alejandro Pascual-Iglesias5Verónica Terrón-Arcos6Verónica Terrón-Arcos7Gema Muñoz-Bartolo8Esteban Frauca9Francisco J. Cueto10Francisco J. Cueto11Cristina Calvo12Cristina Calvo13Laura Córdoba-García14Laura Córdoba-García15Jesús Fernández-Felipe16Jesús Fernández-Felipe17Laura Hurtado-Navarro18Laura Hurtado-Navarro19Julia del Prado-Montero20Julia del Prado-Montero21Gonzalo Sáenz de Santa María-Diez22Gonzalo Sáenz de Santa María-Diez23Daniel Arvelo-Rosario24Daniel Arvelo-Rosario25Paloma Jara26Carlos del Fresno27Eduardo López-Collazo28Eduardo López-Collazo29Eduardo López-Collazo30Eduardo López-Collazo31Eduardo López-Collazo32The Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainDepartment of Paediatric Hepatology, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz Children’s University Hospital, Madrid, SpainDepartment of Paediatric Hepatology, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz Children’s University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainDepartment of Paediatric Hepatology, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz Children’s University Hospital, Madrid, SpainDepartment of Paediatric Hepatology, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz Children’s University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainDepartment of Paediatrics, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz Children’s University Hospital, Madrid, SpainCentre for Biomedical Research Network, Centro de Investigación Biomédica en Red (CIBER) of Infectious Diseases (CIBERINFEC), Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainDepartment of Paediatric Hepatology, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz Children’s University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainThe Innate Immune Response Group, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainTumour Immunology Laboratory, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainBiobank Platform, Instituto de Investigación Hospital Universitario La Paz (IdiPAZ), La Paz University Hospital, Madrid, SpainCentre for Biomedical Research Network, Centro de Investigación Biomédica en Red (CIBER) of Respiratory Diseases (CIBERES), Madrid, SpainBiomedical Department, UNIE Universidad, Madrid, SpainIntroductionPaediatric acute hepatitis of unknown aetiology (PAHUA) has emerged as a global health concern, yet its cause remains unidentified. This study characterises the clinical and immunological profiles of PAHUA to identify reliable immune biomarkers for accurate diagnosis.MethodsSamples from 24 PAHUA patients, 6 children with autoimmune hepatitis (AIH), and 13 healthy paediatric volunteers (HVs) were analysed. Immunophenotyping, soluble immune checkpoints (ICs) and cytokine profiling, and ex vivo immune responses were assessed using spectral flow cytometry. Clustering and logistic regression modelling were used to identify predictive biomarkers.ResultsPAHUA cases frequently presented with gastrointestinal symptoms and liver damage preceding jaundice, with 59% progressing to paediatric acute liver failure (pALF). Adenovirus was detected in only 17.6% of PAHUA patients, suggesting it is unlikely to be the primary causative agent. Antibodies against the SARS-CoV-2 Spike protein were identified in 88.2% of PAHUA patients, as well as in AIH and HV groups, indicating prior exposure. Immunophenotyping, ICs and cytokine profiling, and ex vivo immune revealed distinct immune profiles between PAHUA and non-PAHUA individuals. Furthermore, clustering and logistic regression modelling identified potential predictive biomarkers, including the plasmatic ICs Gal-9 and sTim-3, alongside specific immune cell populations. Notably, a combined Gal-9 and sTim-3 model achieved an AUC of 1.000 in differentiating PAHUA patients from non-PAHUA individuals.ConclusionsDespite the limited cohort analysed, owing to the rarity of cases worldwide, our data provide valuable insights for an accurate, early, and minimally invasive diagnosis of PAHUA. These patients exhibit a distinct immunological profile, with ICs, particularly Gal-9 and sTim-3, showing strong potential as reliable biomarkers.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1599982/fullhepatitisunknown aetiologybiomarkerGal-9sTim-3
spellingShingle Roberto Lozano-Rodríguez
Roberto Lozano-Rodríguez
Loreto Hierro
María José Quiles
Alejandro Pascual-Iglesias
Alejandro Pascual-Iglesias
Verónica Terrón-Arcos
Verónica Terrón-Arcos
Gema Muñoz-Bartolo
Esteban Frauca
Francisco J. Cueto
Francisco J. Cueto
Cristina Calvo
Cristina Calvo
Laura Córdoba-García
Laura Córdoba-García
Jesús Fernández-Felipe
Jesús Fernández-Felipe
Laura Hurtado-Navarro
Laura Hurtado-Navarro
Julia del Prado-Montero
Julia del Prado-Montero
Gonzalo Sáenz de Santa María-Diez
Gonzalo Sáenz de Santa María-Diez
Daniel Arvelo-Rosario
Daniel Arvelo-Rosario
Paloma Jara
Carlos del Fresno
Eduardo López-Collazo
Eduardo López-Collazo
Eduardo López-Collazo
Eduardo López-Collazo
Eduardo López-Collazo
Comprehensive immune profiling and predictive modelling of paediatric acute hepatitis of unknown aetiology from a Spanish cohort
Frontiers in Immunology
hepatitis
unknown aetiology
biomarker
Gal-9
sTim-3
title Comprehensive immune profiling and predictive modelling of paediatric acute hepatitis of unknown aetiology from a Spanish cohort
title_full Comprehensive immune profiling and predictive modelling of paediatric acute hepatitis of unknown aetiology from a Spanish cohort
title_fullStr Comprehensive immune profiling and predictive modelling of paediatric acute hepatitis of unknown aetiology from a Spanish cohort
title_full_unstemmed Comprehensive immune profiling and predictive modelling of paediatric acute hepatitis of unknown aetiology from a Spanish cohort
title_short Comprehensive immune profiling and predictive modelling of paediatric acute hepatitis of unknown aetiology from a Spanish cohort
title_sort comprehensive immune profiling and predictive modelling of paediatric acute hepatitis of unknown aetiology from a spanish cohort
topic hepatitis
unknown aetiology
biomarker
Gal-9
sTim-3
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1599982/full
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