Characterisation of Foxp3 splice variants in human CD4+ and CD8+ T cells—Identification of Foxp3Δ7 in human regulatory T cells
Abstract
Foxp3 is proposed to play a critical role in the development and function of regulatory T cells. Functional and transgenic studies in mice propose Foxp3 as a "regulatory T cell lineage specification factor" but conflicting data exist in humans. Expression of multiple Foxp3 splice variants in humans represents an additional layer of complexity for this transcription factor and acts as a possible mechanism of regulating protein diversity. We report the identification of a novel splice variant of Foxp3, called Foxp3Δ7, in ex vivo CD4+CD25+ T cells and CD8+ regulatory T cell clones. Foxp3Δ7 lacks the 81bp region that encodes exon 7 of Foxp3, which is a part of the leucine zipper domain of the protein. The three splice variants of Foxp3 namely Foxp3FL, Foxp3Δ2 and Foxp3Δ7 are co-expressed in ex vivo human CD4+CD25+ T cells and CD8+ Treg clones. Stimulation of freshly isolated CD4+CD25+ T cells with anti-CD3 and anti-CD28 antibodies leads to a 140-fold upregulation of Foxp3Δ7 within 24h of stimulation, which is ∼10-fold greater than that observed in stimulated CD4+CD25- T cells. In addition, resting CD8+ Treg cells have decreased expression of Foxp3FL and Foxp3Δ2; however they have a 10-fold higher expression of Foxp3Δ7, in comparison to ex vivo CD4+CD25+ T cells. In order to assess the functional effects of these Foxp3 isoforms, we carried out lentivirus expression studies. All three isoforms were capable of inducing increased levels of CD25 expression in primary human CD4+ T cells, along with a tendency to decreased levels of CD127. Further investigation into pathways that alter the relative proportions of Foxp3 isoforms, and hence their interaction with other transcriptional co-regulators, will help to define the role of Foxp3 isoforms in immune regulation.
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There is a general consensus that the 236A/E7 and 259D anti-human FOXP3 Ab clones are optimal.
“Several studies have compared different anti-human FOXP3 Ab clones, and although there is some debate, there is a general consensus that the 236A/E7 and 259D clones are optimal [394-398].”
From Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition) (2021) · cites this paper as [395]
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- Authors
- Kaur, G., Goodall, J. C., Jarvis, L. B., & Hill Gaston, J. S.
- Affiliations
- University of Cambridge (GB)
- Venue
- Molecular Immunology
- Publisher
- Elsevier BV
- Published
- 2010-11
- Volume
- 48
- Issue
- 1-3
- Pages
- 321-332
- ISSN
- 0161-5890 (ISSN-L)
- OpenAlex
- W2065059935
- Cited by
- 45
- References
- 73
- Citations by year
- 2025: 3 · 2024: 3 · 2023: 2 · 2022: 6 · 2021: 1 · 2020: 2 · 2019: 3 · 2018: 3 · 2017: 4 · 2016: 3 · 2015: 5 · 2014: 1 · 2013: 2 · 2012: 3
- FWCI
- 1.24
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Molecular Immunology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Versus Arthritis
- NIHR Cambridge Biomedical Research Centre
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › FOXO transcription factor regulation
- Life Sciences › Immunology and Microbiology › Immunology › Psoriasis: Treatment and Pathogenesis
- Keywords
- Foxp3, regulatory T cells, CD4+CD25+ T cells, CD8+ regulatory T cells, leucine zipper domain, CD25 expression, CD127, T cell activation, transcription factor isoforms, immune regulation
- MeSH terms
- Base Sequence, Cell Separation, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Humans, Lymphocyte Activation, Molecular Sequence Data, Blotting, Western, CD4-Positive T-Lymphocytes, CD8-Positive T-Lymphocytes, Protein Isoforms, Reverse Transcriptase Polymerase Chain Reaction, T-Lymphocytes, Regulatory, Forkhead Transcription Factors
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