Immunophenotyping of sheep paraffin-embedded peripheral lymph nodes

Melzi, E., Rocchi, M. S., Entrican, G., Caporale, M. and Palmarini, M. (2018) Immunophenotyping of sheep paraffin-embedded peripheral lymph nodes. Frontiers in Immunology, 9, 2892. (doi:10.3389/fimmu.2018.02892) (PMID:30619264) (PMCID:PMC6297804)

Melzi, E., Rocchi, M. S., Entrican, G., Caporale, M. and Palmarini, M. (2018) Immunophenotyping of sheep paraffin-embedded peripheral lymph nodes. Frontiers in Immunology, 9, 2892. (doi:10.3389/fimmu.2018.02892) (PMID:30619264) (PMCID:PMC6297804)

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Abstract

Sheep are not only a major livestock species globally, they are also an important large animal model for biomedical research and have contributed to our understanding of the ontogeny and architecture of the mammalian immune system. In this study, we applied immunohistochemistry and multicolor immunofluorescence in fixed and paraffin-embedded lymph nodes to phenotype the key populations of antigen presenting cells, lymphocytes and stromal cells that orchestrate the host adaptive immune response. We used an extensive panel of antibodies directed against markers associated with dendritic cells (MHC class II, CD83 and CD208), macrophages (CD11b, CD163 and CD169), stromal cells (CNA.42, S100 and CD83), and lymphocytes (CD3, Pax5, CD4, CD8). Using different methods of tissue fixation and antigen retrieval, we provide a detailed immunophenotyping of sheep lymph nodes including the identification of potential subpopulations of antigen presenting cells and stromal cells. By characterizing cells expressing combinations of these markers in the context of their morphology and location within the lymph node architecture, we provide valuable new tools to investigate the structure, activation and regulation of the sheep immune system in health and disease.

Item Type:Articles
Additional Information:This study was funded by the Wellcome Trust (Investigator Award # 206369/Z/17/Z) and in part by a W.B. Martin Scholarship. Animal experiments conducted at the Istituto Zooprofilattico Sperimentale dell'Abruzzo e Molise G. Caporale were funded by the Italian Ministry of Health. GE and MSR are funded by the Scottish Government Rural and Environment Science and Analytical Services (RESAS). GE is also funded under European Union's Horizon 2020 research and innovation program under grant agreement No. 731014 (VetBioNet).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Entrican, Professor Gary and Melzi, Miss Eleonora and Caporale, Dr Marco and Palmarini, Professor Massimo and Rocchi, Dr Mara
Authors: Melzi, E., Rocchi, M. S., Entrican, G., Caporale, M., and Palmarini, M.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Frontiers in Immunology
Publisher:Frontiers
ISSN:1664-3224
ISSN (Online):1664-3224
Copyright Holders:Copyright © 2018 Melzi, Rocchi, Entrican, Caporale and Palmarini
First Published:First published in Frontiers in Immunology 9:2892
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
3010490Host determinants of disease outcomes in arboviral infectionsMassimo PalmariniWellcome Trust (WELLCOTR)206369/Z/17/ZIII - Centre for Virus Research