Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.806741
Title: The in vitro transcriptional regulation of interleukin-10 versus pro-inflammatory cytokine gene expression by c-Maf and Blimp-1 in CD4⁺ T-helper cells
Author: Cox, Luke
Awarding Body: King's College London
Current Institution: King's College London (University of London)
Date of Award: 2020
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Abstract:
During infection CD4+ T-helper cells are an important source of the anti-inflammatory cytokine interleukin (IL)-10, which is important in preventing T cell-mediated immunopathology. Despite having different differentiation programs, expressing unique hallmark transcription factors and pro-inflammatory cytokines, all CD4+ T-helper cell subsets can express Il10. Previous RNA-sequencing analysis in the lab identified the transcription factors: c-Maf (encoded by Maf) and B-Lymphocyte-Induced Maturation Protein 1 (Blimp-1, encoded by Prdm1) to positively correlate with Il10 expression across multiple CD4+ T-helper cell subsets. To investigate the role of c-Maf and Blimp-1 on cytokine gene regulation we differentiated multiple CD4+ T-helper cell subsets in vitro and confirmed the peak of Il10 expression, alongside the expression of, Maf, Prdm1, hallmark transcription factors and pro-inflammatory cytokines. This analysis revealed that each CD4+ T-helper cell subset expressed varying levels of, Il10, pro-inflammatory cytokines, Maf and Prdm1. We then assessed the effect of Cd4Cre-mediated deletion of either Maf, Prdm1 or both on the expression of cytokines and transcription factors in the relevant subsets. In TH17-cells, deletion of Maf reduced Il10 and Rorc expression, whilst increasing Il2 expression. Anti-IL-2 treatment revealed that both c-Maf and IL-2 independently promote Il10 expression by TH17-cells, and that c-Maf promotes Rorc expression via its negative regulation of Il2. In contrast, deletion of either Maf or Prdm1 did not affect Il10 expression by TH2-cells. In CD4+ T-helper cells differentiated with IL- 27, deletion of either Maf, Prdm1 or both reduced Il10, whilst increasing Ifng expression, and revealed that c-Maf and Blimp-1 co-regulate each other’s expression. RNAsequencing analysis of IL-27-driven CD4+ T-helper cells revealed c-Maf and Blimp-1 to be the top two transcription factors positively correlating with Il10 expression in these cells, and identified other novel transcription factors correlating with Il10 and Ifng. We also performed RNA-sequencing analysis of IL-27-driven CD4+ T-helper cells with deletion of either Maf or Prdm1, identifying some of the common and distinct transcription factors regulated by c-Maf and Blimp-1. Overall, the work presented in this thesis demonstrates that c-Maf and Blimp-1 are important regulators of Il10 versus Ifng expression by IL-27- driven CD4+ T-helper cells, and provides a basis for further investigation of the molecular mechanisms regulating Il10 versus Ifng expression by these cells.
Supervisor: Hawrylowicz, Catherine Martha Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.806741  DOI: Not available
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