Ex vivo-stimulated cytokine expression in whole blood indicates swine influenza vaccine-induced immune memory

IF 1.8 3区农林科学 Q4 IMMUNOLOGY

Veterinary immunology and immunopathology Pub Date : 2026-01-01 Epub Date: 2025-11-19 DOI:10.1016/j.vetimm.2025.111024

Giulia Fantoni , Andrea Müllebner , Nimród Pálmai , Péter Trampus , Kathrin Lillie-Jaschniski , Han Smits , Navodita Maurice , Krishna Latha , J. Catharina Duvigneau , István Kiss

{"title":"Ex vivo-stimulated cytokine expression in whole blood indicates swine influenza vaccine-induced immune memory","authors":"Giulia Fantoni , Andrea Müllebner , Nimród Pálmai , Péter Trampus , Kathrin Lillie-Jaschniski , Han Smits , Navodita Maurice , Krishna Latha , J. Catharina Duvigneau , István Kiss","doi":"10.1016/j.vetimm.2025.111024","DOIUrl":null,"url":null,"abstract":"<div><div>Swine influenza viruses (SwIAV) pose a zoonotic threat and contribute to interspecies viral reassortment. Effective vaccination strategies are essential, yet accessible correlates of protection remain limited. We simplified a whole blood (WB) assay to assess immune memory ex vivo in pigs vaccinated with Respiporc FLU3® (n = 6) versus placebo controls (n = 6), followed by homologous challenge with SwIAV (H1avN1). Vaccinated pigs exhibited no lung lesions and minimal viral loads. WB samples, collected pre- and post-vaccination and at day 1 and 5 post-challenge (dpch), were stimulated with SwIAV to quantify IL2, IL4, IFNγ, TNFα, and granzyme B (GZMB) mRNA via qPCR. High IL2 and moderate IL4 mRNA elicited in WB post-booster predicted reduced viral load and lung pathology. Concomitantly, high IFNγ in WB from controls at 5dpch correlated with lesion severity, indicating excessive inflammation in non-vaccinates. Post-challenge, IL4/IL2 mRNA ratios increased in virus-stimulated WB of vaccinates, suggesting Th2-associated resolution of infection. In contrast, controls showed declining IL4/IL2 ratios post-challenge, correlating with disease severity. Thus, the simplified WB assay allows identification of virus-specific immune responses associating with protection, without requiring extensive laboratory infrastructure. By recording the ex vivo-triggered SwIAV-specific IL4/IL2 mRNA dynamics, which develop between booster and challenge monitoring immune memory may simplify determination of vaccination success.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"291 ","pages":"Article 111024"},"PeriodicalIF":1.8000,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Veterinary immunology and immunopathology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0165242725001448","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/11/19 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"IMMUNOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Swine influenza viruses (SwIAV) pose a zoonotic threat and contribute to interspecies viral reassortment. Effective vaccination strategies are essential, yet accessible correlates of protection remain limited. We simplified a whole blood (WB) assay to assess immune memory ex vivo in pigs vaccinated with Respiporc FLU3® (n = 6) versus placebo controls (n = 6), followed by homologous challenge with SwIAV (H1avN1). Vaccinated pigs exhibited no lung lesions and minimal viral loads. WB samples, collected pre- and post-vaccination and at day 1 and 5 post-challenge (dpch), were stimulated with SwIAV to quantify IL2, IL4, IFNγ, TNFα, and granzyme B (GZMB) mRNA via qPCR. High IL2 and moderate IL4 mRNA elicited in WB post-booster predicted reduced viral load and lung pathology. Concomitantly, high IFNγ in WB from controls at 5dpch correlated with lesion severity, indicating excessive inflammation in non-vaccinates. Post-challenge, IL4/IL2 mRNA ratios increased in virus-stimulated WB of vaccinates, suggesting Th2-associated resolution of infection. In contrast, controls showed declining IL4/IL2 ratios post-challenge, correlating with disease severity. Thus, the simplified WB assay allows identification of virus-specific immune responses associating with protection, without requiring extensive laboratory infrastructure. By recording the ex vivo-triggered SwIAV-specific IL4/IL2 mRNA dynamics, which develop between booster and challenge monitoring immune memory may simplify determination of vaccination success.

查看原文本刊更多论文

全血中体外刺激的细胞因子表达表明猪流感疫苗诱导的免疫记忆

猪流感病毒（SwIAV）造成人畜共患威胁，并有助于种间病毒重组。有效的疫苗接种战略至关重要，但可获得的相关保护仍然有限。我们简化了一种全血（WB）测定方法，以评估接种了Respiporc FLU3®（n = 6）和安慰剂对照（n = 6）的猪的体外免疫记忆，然后用猪流感病毒（H1avN1）进行同源攻击。接种疫苗的猪没有肺部病变，病毒载量也很低。接种前和接种后以及攻毒后第1天和第5天收集WB样本，用SwIAV刺激，通过qPCR定量IL2、IL4、IFNγ、TNFα和颗粒酶B (GZMB) mRNA。高il - 2和中等il - 4 mRNA在WB增强后的表达预示着病毒载量和肺病理的降低。同时，5dpch时WB中IFNγ含量高与病变严重程度相关，表明未接种疫苗者炎症过度。攻毒后，疫苗中病毒刺激的WB中IL4/IL2 mRNA比值增加，提示th2相关的感染解决。相比之下，对照组在攻击后显示IL4/IL2比率下降，与疾病严重程度相关。因此，简化的WB测定法可以识别与保护相关的病毒特异性免疫反应，而不需要大量的实验室基础设施。通过记录在体外触发的swiav特异性IL4/IL2 mRNA动态，监测免疫记忆可以简化疫苗接种成功的确定。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Veterinary immunology and immunopathology 农林科学-免疫学

CiteScore

3.40

自引率

5.60%

发文量

审稿时长

70 days

期刊介绍： The journal reports basic, comparative and clinical immunology as they pertain to the animal species designated here: livestock, poultry, and fish species that are major food animals and companion animals such as cats, dogs, horses and camels, and wildlife species that act as reservoirs for food, companion or human infectious diseases, or as models for human disease. Rodent models of infectious diseases that are of importance in the animal species indicated above,when the disease requires a level of containment that is not readily available for larger animal experimentation (ABSL3), will be considered. Papers on rabbits, lizards, guinea pigs, badgers, armadillos, elephants, antelope, and buffalo will be reviewed if the research advances our fundamental understanding of immunology, or if they act as a reservoir of infectious disease for the primary animal species designated above, or for humans. Manuscripts employing other species will be reviewed if justified as fitting into the categories above. The following topics are appropriate: biology of cells and mechanisms of the immune system, immunochemistry, immunodeficiencies, immunodiagnosis, immunogenetics, immunopathology, immunology of infectious disease and tumors, immunoprophylaxis including vaccine development and delivery, immunological aspects of pregnancy including passive immunity, autoimmuity, neuroimmunology, and transplanatation immunology. Manuscripts that describe new genes and development of tools such as monoclonal antibodies are also of interest when part of a larger biological study. Studies employing extracts or constituents (plant extracts, feed additives or microbiome) must be sufficiently defined to be reproduced in other laboratories and also provide evidence for possible mechanisms and not simply show an effect on the immune system.