Development of monoclonal antibodies to detect bovine FOXP3 in PBMCs exposed to a staphylococcal superantigen

K.S Seo; W.C Davis; M.J Hamilton; Y.H Park; G.A Bohach

doi:10.1016/j.vetimm.2008.10.292

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Development of monoclonal antibodies to detect bovine FOXP3 in PBMCs exposed to a staphylococcal superantigen

Journal article

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Development of monoclonal antibodies to detect bovine FOXP3 in PBMCs exposed to a staphylococcal superantigen

K.S Seo, W.C Davis, M.J Hamilton, Y.H Park and G.A Bohach

Veterinary immunology and immunopathology, Vol.128(1-3), pp.30-36

03/15/2009

DOI: https://doi.org/10.1016/j.vetimm.2008.10.292

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https://hdl.handle.net/2376/102985

PMCID: PMC3774054

PMID: 19054574

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Abstract

Superantigen

Bovine FOXP3

Staphylococcal

TCR Vβ

Treg

The role of regulatory T cells (Tregs) is well documented in immune homeostasis and protection against autoimmune disease. Forkhead box protein 3 (FOXP3) has been shown to be essential for the development and function of Treg. Due to the lack of tools for FOXP3 detection in certain species, understanding the role of Treg in a variety of ruminant diseases has been hampered. In this study, we developed monoclonal antibodies (mAbs) against bovine FOXP3 using recombinant bovine FOXP3 lacking the forkhead domain as an immunogen. The specificity of the mAbs was confirmed by immunoblot and mass spectrometry. Expression of FOXP3 was induced in bovine PBMCs after 6 d of exposure to staphylococcal enterotoxin type C1 (SEC1) in vitro. Similar to findings in mice and humans, expression of FOXP3 was restricted to CD4+ CD25+ T cells. Transcriptional analysis of bovine TCR variable regions of the β chain (boVβ) showed that transcription of boVβ sequences reactive with SEC1 increased for 6 d, and then boVβ sequences non-reactive with SEC1 rapidly increased in the cultures. This indicates that induction of FOXP3+ CD4+ CD25+ Tregs by SEC1 is not Vβ restricted. The FOXP3 mAbs developed in this study will be useful in the further investigation of the role of Treg in staphylococcal pathogenesis in bovine mastitis and other ruminant diseases.

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Title: Development of monoclonal antibodies to detect bovine FOXP3 in PBMCs exposed to a staphylococcal superantigen
Creators: K.S Seo - Department of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow, ID 83844, United States
W.C Davis - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164, United States
M.J Hamilton - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164, United States
Y.H Park - Department of Microbiology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Seoul 151-742, South Korea
G.A Bohach - Department of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow, ID 83844, United States
Publication Details: Veterinary immunology and immunopathology, Vol.128(1-3), pp.30-36
Academic Unit: Veterinary Microbiology and Pathology, Department of
Publisher: Elsevier B.V
Identifiers: 99900546548601842
Language: English
Resource Type: Journal article