Diverse Inflammatory Cytokines Induce Selectin Ligand Expression on Murine CD4 T Cells via p38α MAPK

Ebel, Mark E.; Awe, Olufolakemi; Kaplan, Mark H.; Kansas, Geoffrey S.

doi:10.4049/jimmunol.1500485

Cited by 21 publications

(25 citation statements)

References 70 publications

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“…1I) (20), and this is also observed with other cytokines (20). The basis for this pattern of expression has never been determined, but could potentially be explained by a threshold of TGF-b1 signal strength required for induction of selectin ligands.…”

Section: Resultsmentioning

confidence: 83%

“…Loss of both Smad2 and Smad3 in conventional T cells leads them to be refractory to TGF-b1-mediated inhibition, leading to similar pathology (10,21). In addition to a role in Th cell differentiation, TGF-b1 is a potent inducer of E-and P-selectin ligands in CD4 T cells, both in humans and mice (19,20). We have recently shown that this induction of selectin ligands requires p38a MAPK (20), one of several non-Smad pathways activated by TGFb1 (13,14).…”

Section: Discussionmentioning

confidence: 99%

“…Isolation of splenic CD4 T cells, activation by plate-bound anti-CD3/C28, each at 1 mg/ml for all experiments, culture in IL-2 with and without 5 ng/ml TGF-b1, analysis of E-and P-selectin ligand expression by flow cytometry using E-and P-selectin/IgM chimeras, retroviral transduction of activated CD4 T cells, isolation of RNA, and quantitative RT-PCR were all exactly as recently described (20). Intracellular staining for IL-17, IL-9, and Foxp3, using PE-conjugated (IL-9 and Foxp3) or PerCP-conjugated (IL-17) mAbs from eBioscience, was performed by standard methods following culture under Th17 (TGF-b1 plus IL-6), Th9 (TGF-b1 plus IL-4 plus IL-2), or induced Treg (iTreg; TGF-b1 + IL-2) conditions, as described (20).…”

Section: T Cell Culture and Analysismentioning

confidence: 99%

“…Intracellular staining for IL-17, IL-9, and Foxp3, using PE-conjugated (IL-9 and Foxp3) or PerCP-conjugated (IL-17) mAbs from eBioscience, was performed by standard methods following culture under Th17 (TGF-b1 plus IL-6), Th9 (TGF-b1 plus IL-4 plus IL-2), or induced Treg (iTreg; TGF-b1 + IL-2) conditions, as described (20). For Th17 and Th9, cells were restimulated separately with plate-bound anti-CD3 or PMA/ionomycin prior to addition of GolgiStop, with substantially similar results.…”

Section: T Cell Culture and Analysismentioning

confidence: 99%

“…Selectin ligand expression on activated T cells is controlled largely by cytokines (17)(18)(19). We have recently identified a biologically diverse set of Th cell-promoting and inflammatory cytokines, including TGF-b1, that strongly induce expression of selectin ligands, and each of which requires p38a MAPK for induction of selectin ligands (20). However, a potential role for Smad transcription factors in TGF-b1-induced selectin ligand induction has never been assessed, despite the clear importance of the TGF-b1/Smad pathway to most biological responses to TGF-b1, as well as to regulation of numerous aspects of immunity, including Th cell differentiation.…”

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confidence: 99%

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Functions of Smad Transcription Factors in TGF-β1–Induced Selectin Ligand Expression on Murine CD4 Th Cells

Ebel

Kansas

2016

The Journal of Immunology

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Selectins are carbohydrate-binding adhesion molecules that control leukocyte traffic. Induction of selectin ligands on T cells is controlled primarily by cytokines, including TGF-β1, and requires p38α MAPK, but transcriptional mechanisms that underlie cytokine-driven selectin ligand expression are poorly understood. In this study, we show, using mice with conditional deletions of the TGF-β1–responsive transcription factors Smad2, Smad3, or Smad4, that induction of selectin ligands on CD4 cells in response to TGF-β1 requires Smad4 plus either Smad2 or Smad3. Analysis of CD4 cells from mice with only one functional Smad4 allele revealed a sharp gene dosage effect, suggesting the existence of a threshold of TGF-β1 signal strength required for selectin ligand induction. Both Smad4 plus either Smad2 or Smad3 were selectively required for induction of Fut7 and Gcnt1, glycosyltransferases critical for selectin ligand biosynthesis, but they were not required for St3gal4 or St3gal6 induction. Smad4 plus either Smad2 or Smad3 were also required for induction of Runx transcription factors by TGF-β1. Enforced expression of Runx2, but not Runx1 or Runx3, in Smad2/Smad3 doubly deficient CD4 cells restored selectin ligand expression to wild-type levels. In contrast, enforced expression of Runx1, Runx2, or Runx3 failed to restore differentiation of TGF-β1–dependent Th cell lineages, including Th17, Th9, and induced regulatory T cells. These results show that Smads are directly required for Th cell differentiation independent of Runx induction but only indirectly required via Runx2 for TGF-β1–induced selectin ligand induction on murine CD4 T cells.

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Section: Resultsmentioning

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Section: T Cell Culture and Analysismentioning

confidence: 99%

Section: T Cell Culture and Analysismentioning

confidence: 99%

mentioning

confidence: 99%

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Functions of Smad Transcription Factors in TGF-β1–Induced Selectin Ligand Expression on Murine CD4 Th Cells

Ebel

Kansas

2016

The Journal of Immunology

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TCR signalling network organization at the immunological synapses of murine regulatory T cells

et al. 2017

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Regulatory T (Treg) cells require T-cell receptor (TCR) signalling to exert their immunosuppressive activity, but the precise organization of the TCR signalling network compared to conventional T (Tconv) cells remains elusive. By using accurate mass spectrometry and multi-epitope ligand cartography (MELC) we characterized TCR signalling and recruitment of TCR signalling components to the immunological synapse (IS) in www.eji-journal.euThis is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. 2Marco van Ham et al. Eur. J. Immunol. 2017. 0: 1-16 in Treg cells, thereby, seems to be differentially organized as in Tconv cells: Treg cells show reduced Ca 2+ flux and ERK1/2 phosphorylation upon TCR stimulation [6][7][8], and downstream signalling molecules such as Lck, LAT and PLCγ1 are essential for Treg cell suppressive capacity [9,10]. In this line, a novel TCRmediated ADAP/integrin-independent PLCγ1 activation pathway was described to be required for suppression of Tconv cells by Treg cells [11]. Furthermore, Treg cells exhibit reduced S473 phosphorylation of Akt which seems a prerequisite for suppression [12]. Although the enzymatic activity of the tyrosine kinase ZAP70 seems to be dispensable for the suppressive phenotype of Foxp3 + Treg cells [13], a single mutation within the SH2-domain of ZAP70 leads to impaired suppressive activities, which indicated its importance at the immunological synapse (IS) [14]. Finally, it has only recently been recognized that the recruitment of TCR signalling components into the IS is differentially organized in Treg cells and Tconv cells. The protein kinase PKCθ is recruited to the IS in Tconv cells, in Treg cells, however, this kinase was found to be sequestered away from the IS, but still of importance to control suppression [15]. The spatial recruitment of signalling components during IS formation critically depends on cytoskeleton dynamics, which in turn is controlled by TCR activation and subsequent phosphorylation of proteins regulating cytoskeleton reorganization [5]. As part of these processes the microtubule-organizing centre (MTOC) is rapidly translocated in proximity to the IS. MTOC repositioning depends on LAT, ZAP70 and SLP76 [16] and is regulated by a cascade of distinct isoforms of the family of novel protein kinase C (nPKC) [17]. The MTOC serves as a platform to coordinate molecular movements from and to the IS through the support of microtubule (MT) motors [18]. For instance, TCR microclusters move along MTs towards the centre of the IS in a dynein-dependent manner [19], and hindrance of MTOC polarization, molecular transport and cytoskeleton dynamics prevent proper propagation of TCR signals [20].It is now tempting to speculate that the localization of signalling modules within the IS may be instrumental for the formation of a Treg cell-specific IS and the suppressive phenotype of Treg cells. At this moment, however,...

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Glycans as Targets and Mediators of T-Cell Immunotherapy

Wellhausen

Aguilar

Posey

2021

Comprehensive Glycoscience

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Diverse Inflammatory Cytokines Induce Selectin Ligand Expression on Murine CD4 T Cells via p38α MAPK

Cited by 21 publications

References 70 publications

Functions of Smad Transcription Factors in TGF-β1–Induced Selectin Ligand Expression on Murine CD4 Th Cells

Functions of Smad Transcription Factors in TGF-β1–Induced Selectin Ligand Expression on Murine CD4 Th Cells

TCR signalling network organization at the immunological synapses of murine regulatory T cells

Glycans as Targets and Mediators of T-Cell Immunotherapy

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