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Application of the TruCulture® whole blood stimulation system for immune response profiling in cattle.
Vet Immunol Immunopathol. 2020 Mar; 221:110025.VI

Abstract

Capturing the phenotypic variation in immune responses holds enormous promise for the development of targeted treatments for disease as well as tailored vaccination schedules. However, accurate detection of true biological variation can be obscured by the lack of standardised immune assays. The TruCulture® whole blood stimulation system has now been extensively used to detect basal and induced immune responses to a range of pathogen-associated molecular patterns (PAMPs) in human peripheral blood. This study demonstrates the optimisation of this commercially available assay for systemic immune phenotyping in cattle. The early immune response in Holstein-Friesian bull calves (n = 10) was assessed by haematology, flow cytometry and cytokine expression profiling after 24 h ex-vivo PAMP (LPS, poly (I:C) and zymosan) stimulation in TruCulture® tubes. A comparative analysis was also performed with a traditional whole blood stimulation assay and cell viability using both systems was also evaluated.

Results:

Supernatant collected from TruCulture® tubes showed a significant increase in IL-1β and IL-8 expression compared to null stimulated tubes in response to both LPS and zymosan. In contrast, a detectable immune response was not apparent at the standard concentration of poly (I:C). Conventional whole blood cultures yielded similar response profiles, although the magnitude of the response was higher to both LPS and zymosan, which may be attributed to prokaryotic strain-specificity or batch of the stimulant used. Despite being a closed system, HIF1A expression - used as a measure of hypoxia was not increased, suggesting the TruCulture® assay did not negatively affect cell viability. This represents the first reported use of this novel standardised assay in cattle, and indicates that the concentration of poly (I:C) immunogenic in humans is insufficient to induce cytokine responses in cattle. We conclude that the low blood volume and minimally invasive TruCulture® assay system offers a practical and informative technique to assess basal and induced systemic immune responses in cattle.

Authors+Show Affiliations

Animal & Bioscience Research Department, Animal & Grassland Research and Innovation Centre, Teagasc, Grange, Co Meath, Ireland; Host-Pathogen Interactions Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin 2, Ireland.Host-Pathogen Interactions Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin 2, Ireland.Animal & Bioscience Research Department, Animal & Grassland Research and Innovation Centre, Teagasc, Grange, Co Meath, Ireland. Electronic address: kieran.meade@teagasc.ie.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32086040

Citation

O'Brien, Megan B., et al. "Application of the TruCulture® Whole Blood Stimulation System for Immune Response Profiling in Cattle." Veterinary Immunology and Immunopathology, vol. 221, 2020, p. 110025.
O'Brien MB, McLoughlin RM, Meade KG. Application of the TruCulture® whole blood stimulation system for immune response profiling in cattle. Vet Immunol Immunopathol. 2020;221:110025.
O'Brien, M. B., McLoughlin, R. M., & Meade, K. G. (2020). Application of the TruCulture® whole blood stimulation system for immune response profiling in cattle. Veterinary Immunology and Immunopathology, 221, 110025. https://doi.org/10.1016/j.vetimm.2020.110025
O'Brien MB, McLoughlin RM, Meade KG. Application of the TruCulture® Whole Blood Stimulation System for Immune Response Profiling in Cattle. Vet Immunol Immunopathol. 2020;221:110025. PubMed PMID: 32086040.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Application of the TruCulture® whole blood stimulation system for immune response profiling in cattle. AU - O'Brien,Megan B, AU - McLoughlin,Rachel M, AU - Meade,Kieran G, Y1 - 2020/02/11/ PY - 2019/07/08/received PY - 2020/02/05/revised PY - 2020/02/10/accepted PY - 2020/2/23/pubmed PY - 2020/2/23/medline PY - 2020/2/23/entrez KW - Bovine peripheral blood KW - Innate immune response KW - PAMPs KW - TruCulture® SP - 110025 EP - 110025 JF - Veterinary immunology and immunopathology JO - Vet. Immunol. Immunopathol. VL - 221 N2 - Capturing the phenotypic variation in immune responses holds enormous promise for the development of targeted treatments for disease as well as tailored vaccination schedules. However, accurate detection of true biological variation can be obscured by the lack of standardised immune assays. The TruCulture® whole blood stimulation system has now been extensively used to detect basal and induced immune responses to a range of pathogen-associated molecular patterns (PAMPs) in human peripheral blood. This study demonstrates the optimisation of this commercially available assay for systemic immune phenotyping in cattle. The early immune response in Holstein-Friesian bull calves (n = 10) was assessed by haematology, flow cytometry and cytokine expression profiling after 24 h ex-vivo PAMP (LPS, poly (I:C) and zymosan) stimulation in TruCulture® tubes. A comparative analysis was also performed with a traditional whole blood stimulation assay and cell viability using both systems was also evaluated. Results: Supernatant collected from TruCulture® tubes showed a significant increase in IL-1β and IL-8 expression compared to null stimulated tubes in response to both LPS and zymosan. In contrast, a detectable immune response was not apparent at the standard concentration of poly (I:C). Conventional whole blood cultures yielded similar response profiles, although the magnitude of the response was higher to both LPS and zymosan, which may be attributed to prokaryotic strain-specificity or batch of the stimulant used. Despite being a closed system, HIF1A expression - used as a measure of hypoxia was not increased, suggesting the TruCulture® assay did not negatively affect cell viability. This represents the first reported use of this novel standardised assay in cattle, and indicates that the concentration of poly (I:C) immunogenic in humans is insufficient to induce cytokine responses in cattle. We conclude that the low blood volume and minimally invasive TruCulture® assay system offers a practical and informative technique to assess basal and induced systemic immune responses in cattle. SN - 1873-2534 UR - https://www.unboundmedicine.com/medline/citation/32086040/Application_of_the_TruCulture®_whole_blood_stimulation_system_for_immune_response_profiling_in_cattle L2 - https://linkinghub.elsevier.com/retrieve/pii/S0165-2427(19)30242-9 DB - PRIME DP - Unbound Medicine ER -
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