Multiparameter flow cytometry assay to analyze the pulmonary T cell profiles in the ovine model of respiratory syncytial virus infection

Démoulins, Thomas; Brügger, Melanie; Zumkehr, Béatrice; Esteves, Blandina I. Oliveira; Ruggli, Nicolas; Alves, Marco P.

doi:10.1016/j.xpro.2022.101688

Cited by 2 publications

(1 citation statement)

References 3 publications

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“…The different bovine immune cell subtypes were identified by FCM using a 7-step, 11–12-color staining protocols, based on recent reports [ 26 , 27 ]. All antibodies used for the procedure, as well as their clones, host, working dilution, and references, are listed in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

Temperature impacts the bovine ex vivo immune response towards Mycoplasmopsis bovis

Démoulins,

Yimthin,

Lindtke

et al. 2024

Vet Res

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Although cattle are the mammalian species with most global biomass associated with a huge impact on our planet, their immune system remains poorly understood. Notably, the bovine immune system has peculiarities such as an overrepresentation of γδ T cells that requires particular attention, specifically in an infectious context. In line of 3R principles, we developed an ex vivo platform to dissect host–pathogen interactions. The experimental design was based on two independent complementary readouts: firstly, a novel 12–14 color multiparameter flow cytometry assay measuring maturation (modulation of cell surface marker expression) and activation (intracellular cytokine detection) of monocytes, conventional and plasmacytoid dendritic cells, natural killer cells, γδ T cells, B and T cells; secondly, a multiplex immunoassay monitoring bovine chemokine and cytokine secretion levels. The experiments were conducted on fresh primary bovine blood cells exposed to Mycoplasmopsis bovis (M. bovis), a major bovine respiratory pathogen. Besides reaffirming the tight cooperation of the different primary blood cells, we also identified novel key players such as strong IFN-γ secreting NK cells, whose role was so far largely overlooked. Additionally, we compared the host–pathogen interactions at different temperatures, including commonly used 37 °C, ruminant body temperature (38–38.5 °C) and fever (≥ 39.5 °C). Strikingly, working under ruminant physiological temperature influenced the capacity of most immune cell subsets to respond to M. bovis compared to 37 °C. Under fever-like temperature conditions the immune response was impaired compared to physiological temperature. Our experimental approach, phenotypically delineating the bovine immune system provided a thorough vision of the immune response towards M. bovis and the influence of temperature towards that immune response.

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Section: Methodsmentioning

confidence: 99%

Temperature impacts the bovine ex vivo immune response towards Mycoplasmopsis bovis

Démoulins,

Yimthin,

Lindtke

et al. 2024

Vet Res

Self Cite

View full text Add to dashboard Cite

show abstract

Temperature effects on the magnitude of the bovineex vivoimmune response towardsMycoplasmopsis bovis

Démoulins,

Yimthin,

Lindtke

et al. 2023

Preprint

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Although cattle are the mammalian species with most global biomass associated with a huge impact on our planet, their immune system remains poorly understood. Notably, the bovine immune system has peculiarities such as an overrepresentation of γδ T cells that requires particular attention, specifically in an infectious context. In line of 3R principles, we developed an ex vivo platform to dissect host-pathogen interactions. The experimental design was based on two independent complementary readouts: firstly, a novel 12-14 color multiparameter flow cytometry assay measuring maturation (modulation of cell surface marker expression) and activation (intracellular cytokine detection) of monocytes, conventional and plasmacytoid dendritic cells, natural killer cells, γδ T cells, B and T cells; secondly, a multiplex immunoassay monitoring bovine chemokine and cytokine secretion levels. The experiments were conducted on fresh primary bovine blood cells exposed toMycoplasmopsis bovis(M. bovis), a major bovine respiratory pathogen. Besides reaffirming the tight cooperation of the different primary blood cells, we also identified novel key players such as strong IFN-γ secreting NK cells, whose role was so far largely overlooked. Additionally, we compared the host-pathogen interactions at different temperatures, including commonly used 37 °C, ruminant body temperature (38-38.5 °C) and fever (≥39.5 °C). Strikingly, working under ruminant physiological temperature influenced the capacity of most immune cell subsets to respond toM. boviscompared to 37 °C. Under fever-like temperature conditions the immune response was impaired compared to physiological temperature. Our experimental approach, phenotypically delineating the bovine immune system provided a thorough vision of the immune response towardsM. bovisand the influence of temperature towards that immune response.

show abstract

Multiparameter flow cytometry assay to analyze the pulmonary T cell profiles in the ovine model of respiratory syncytial virus infection

Cited by 2 publications

References 3 publications

Temperature impacts the bovine ex vivo immune response towards Mycoplasmopsis bovis

Temperature impacts the bovine ex vivo immune response towards Mycoplasmopsis bovis

Temperature effects on the magnitude of the bovineex vivoimmune response towardsMycoplasmopsis bovis

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