Mapping Rora expression in resting and activated CD4+ T cells

The transcription factor Rora has been shown to be important for the development of ILC2 and the regulation of ILC3, macrophages and Treg cells. Here we investigate the role of Rora across CD4+ T cells in general, but with an emphasis on Th2 cells, both in vitro as well as in the context of several in vivo type 2 infection models. We dissect the function of Rora using overexpression and a CD4-conditional Rora-knockout mouse, as well as a RORA-reporter mouse. We establish the importance of Rora in CD4+ T cells for controlling lung inflammation induced by Nippostrongylus brasiliensis infection, and have measured the effect on downstream genes using RNA-seq. Using a systematic stimulation screen of CD4+ T cells, coupled with RNA-seq, we identify upstream regulators of Rora, most importantly IL-33 and CCL7. Our data suggest that Rora is a negative regulator of the immune system, possibly through several downstream pathways, and is under control of the local microenvironment.

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Rora regulates activated T helper cells during inflammation

10.1101/709998 ◽

2019 ◽

Cited By ~ 3

Author(s):

Liora Haim-Vilmovsky ◽

Johan Henriksson ◽

Jennifer A Walker ◽

Zhichao Miao ◽

Eviatar Natan ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Treg Cells ◽

Negative Regulator ◽

Nippostrongylus Brasiliensis ◽

Rna Seq ◽

Infection Models

AbstractThe transcription factor Rora has been shown to be important for the development of ILC2 and the regulation of ILC3, macrophages and Treg cells. Here we investigate the role of Rora across CD4+ T cells, both in vitro as well as in the context of several in vivo type 2 infection models. We dissect the function of Rora using overexpression and a CD4-conditional Rora-knockout mouse, as well as a RORA-reporter mouse. We establish the importance of Rora in CD4+ T cells for controlling lung inflammation induced by Nippostrongylus brasiliensis infection, and have measured the effect on downstream genes using RNA-seq. Using a systematic stimulation screen of CD4+ T cells, coupled with RNA-seq, we identify upstream regulators of Rora, most importantly IL-33 and CCL7. Our data suggest that Rora is a negative regulator of the immune system, possibly through several downstream pathways, and is under control of the local microenvironment.

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T-cell–intrinsic Tif1α/Trim24 regulates IL-1R expression on TH2 cells and TH2 cell-mediated airway allergy

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1522287113 ◽

2016 ◽

Vol 113 (5) ◽

pp. E568-E576 ◽

Cited By ~ 10

Author(s):

Jimena Perez-Lloret ◽

Isobel S. Okoye ◽

Riccardo Guidi ◽

Yashaswini Kannan ◽

Stephanie M. Coomes ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Allergic Diseases ◽

Treg Cells ◽

Th2 Cells ◽

T Helper 2 ◽

Cytokines And Chemokines ◽

Th2 Cell

There is a paucity of new therapeutic targets to control allergic reactions and forestall the rising trend of allergic diseases. Although a variety of immune cells contribute to allergy, cytokine-secreting αβ+CD4+ T-helper 2 (TH2) cells orchestrate the type-2–driven immune response in a large proportion of atopic asthmatics. To identify previously unidentified putative targets in pathogenic TH2 cells, we performed in silico analyses of recently published transcriptional data from a wide variety of pathogenic TH cells [Okoye IS, et al. (2014) Proc Natl Acad Sci USA 111(30):E3081–E3090] and identified that transcription intermediary factor 1 regulator-alpha (Tif1α)/tripartite motif-containing 24 (Trim24) was predicted to be active in house dust mite (HDM)- and helminth-elicited Il4gfp+αβ+CD4+ TH2 cells but not in TH1, TH17, or Treg cells. Testing this prediction, we restricted Trim24 deficiency to T cells by using a mixed bone marrow chimera system and found that T-cell–intrinsic Trim24 is essential for HDM-mediated airway allergy and antihelminth immunity. Mechanistically, HDM-elicited Trim24−/− T cells have reduced expression of many TH2 cytokines and chemokines and were predicted to have compromised IL-1–regulated signaling. Following this prediction, we found that Trim24−/− T cells have reduced IL-1 receptor (IL-1R) expression, are refractory to IL-1β–mediated activation in vitro and in vivo, and fail to respond to IL-1β–exacerbated airway allergy. Collectively, these data identify a previously unappreciated Trim24-dependent requirement for IL-1R expression on TH2 cells and an important nonredundant role for T-cell–intrinsic Trim24 in TH2-mediated allergy and antihelminth immunity.

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Defective IL-10 signaling in hyper-IgE syndrome results in impaired generation of tolerogenic dendritic cells and induced regulatory T cells

Journal of Experimental Medicine ◽

10.1084/jem.20100799 ◽

2011 ◽

Vol 208 (2) ◽

pp. 235-249 ◽

Cited By ~ 77

Author(s):

Masako Saito ◽

Masayuki Nagasawa ◽

Hidetoshi Takada ◽

Toshiro Hara ◽

Shigeru Tsuchiya ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Regulatory T Cells ◽

Cd4 T Cells ◽

Elevated Serum ◽

Treg Cells ◽

Th2 Cells ◽

Hyper Ige Syndrome ◽

And Function

Hyper-IgE syndrome (HIES) is a primary immunodeficiency characterized by recurrent staphylococcal infections and atopic dermatitis associated with elevated serum IgE levels. Although defective differentiation of IL-17–producing CD4+ T cells (Th17) partly accounts for the susceptibility to staphylococcal skin abscesses and pneumonia, the pathogenesis of atopic manifestations in HIES still remains an enigma. In this study, we examined the differentiation and function of Th1, Th2, regulatory T cells (Treg cells), and dendritic cells (DCs) in HIES patients carrying either STAT3 or TYK2 mutations. Although the in vitro differentiation of Th1 and Th2 cells and the number and function of Treg cells in the peripheral blood were normal in HIES patients with STAT3 mutations, primary and monocyte-derived DCs showed defective responses to IL-10 and thus failed to become tolerogenic. When treated with IL-10, patient DCs showed impaired up-regulation of inhibitory molecules on their surface, including PD-L1 and ILT-4, compared with control DCs. Moreover, IL-10–treated DCs from patients displayed impaired ability to induce the differentiation of naive CD4+ T cells to FOXP3+ induced Treg cells (iTreg cells). These results suggest that the defective generation of IL-10–induced tolerogenic DCs and iTreg cells may contribute to inflammatory changes in HIES.

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In Vivo Priming of Cd4 T Cells That Produce Interleukin (Il)-2 but Not IL-4 or Interferon (Ifn)-γ, and Can Subsequently Differentiate into IL-4–Or IFN-γ–Secreting Cells

Journal of Experimental Medicine ◽

10.1084/jem.194.8.1069 ◽

2001 ◽

Vol 194 (8) ◽

pp. 1069-1080 ◽

Cited By ~ 91

Author(s):

Xiaowen Wang ◽

Tim Mosmann

Keyword(s):

T Cells ◽

Immune Responses ◽

Cd4 T Cells ◽

Transforming Growth Factor ◽

Cell Receptor ◽

Th2 Cells ◽

Effector Cells ◽

Ifn Γ

The differentiation of antigen-stimulated naive CD4 T cells into T helper (Th)1 or Th2 effector cells can be prevented in vitro by transforming growth factor (TGF)-β and anti–interferon (IFN)-γ. These cells proliferate and synthesize interleukin (IL)-2 but not IFN-γ or IL-4, and can differentiate into either Th1 or Th2 cells. We have now used two-color Elispots to reveal substantial numbers of primed cells producing IL-2 but not IL-4 or IFN-γ during the Th1- or Th2-biased immune responses induced by soluble proteins or with adjuvants. These cells were CD4+CD44high and were present during immediate and long-term immune responses of normal mice. Naive T cell receptor for antigen (TCR) transgenic (DO11.10) T cells were primed in vivo after adoptive transfer into normal hosts and FACS® cloned under conditions that did not allow further differentiation. After clonal proliferation, aliquots of each clone were cultured in Th1- or Th2-inducing conditions. Many in vivo–primed cells were uncommitted, secreting IL-2 but not IL-4 or IFN-γ at the first cloning step, but secreting either IL-4 or IFN-γ after differentiation in the appropriate conditions. These in vivo-primed, uncommitted, IL-2–producing cells may constitute an expanded pool of antigen-specific cells that provide extra flexibility for immune responses by differentiating into Th1 or Th2 phenotypes later during the same or subsequent immune responses.

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In Vivo Activated CD103+CD4+ Regulatory T Cells Ameliorate Ongoing Chronic GVHD.

Blood ◽

10.1182/blood.v110.11.350.350 ◽

2007 ◽

Vol 110 (11) ◽

pp. 350-350

Author(s):

Dongchang Zhao ◽

Chunyan Zhang ◽

Tangsheng Yi ◽

Chia-Lei Lin ◽

Ivan Todorov ◽

...

Keyword(s):

T Cells ◽

B Cells ◽

Cd4 T Cells ◽

Chronic Gvhd ◽

Treg Cells ◽

Dependent Manner ◽

T And B Cells ◽

Ifn Γ

Abstract The αEβ7 integrin CD103 is an excellent marker for identifying in vivo activated CD4+ regulatory T (Treg) cells. CD103− naive Treg cells from donors are effective in prevention but ineffective in treatment of graft versus host disease (GVHD). It is unknown whether in vivo activated donor CD103+ Treg cells can effectively treat ongoing GVHD. We have recently reported a new chronic GVHD model, in which donor DBA/2 (H-2d) spleen cells were transferred to the irradiated BALB/c (H-2d) recipients. The recipients showed chronic GVHD-like syndrome with high-levels of serum autoantibodies, proteinuria, and hair-loss, and the disease induction required both donor CD4+ T and B cells in a cell dose dependent manner (Blood, 2006). In the current studies, we observed that the percentage of CD103+ cells among donor CD4+ T cells in the recipients without disease was up to 45% and was more than two fold higher than that of the recipients with disease. The CD103+CD4+ T cells were more than 97% FoxP3+, which was similar to natural CD25hi Treg cells, but the former expressed markedly higher levels of CCR5 as compared to the latter. The CD103+ Treg cells showed markedly stronger suppression capacity as compared to freshly isolated as well as in vitro anti-CD3-activated CD25hi natural Treg cells. When injected into ongoing chronic GVHD recipients with severe proteinuria, CD103+ Treg cells (1x106) reversed proteinuria and tissue damages in 92% (11/12) of the recipients, and the recipients survived for more than 100 days. In contrast, the in vitro activated natural Treg cells reversed disease in only 25% (2/8) of the recipients, and the freshly isolated CD25hi natural Treg cells reversed none (0/12), and the treated recipients died within 45 days, which was similar to control PBS treated recipients. Furthermore, we found that infusion of the CD103+ Treg cells significantly reduced CD138+ antibody-secreting plasma cells in spleen and reduced serum levels of autoantibodies; and that infusion of the CD103+ Treg cells also significantly reduced CD44loCD62LhiSCA-1hi post-mitotic CD4+ T memory stem cells in spleen as well as pathogenic IFN-γ+CD4+ T cells in liver and nephritogenic IFN-γ+IL-10+CD4+ T cells in kidney tissues. Our results indicate that, different from CD103− naive natural Treg cells that prevent GVHD via suppressing donor T cell activation and expansion, CD103+ Treg cells ameliorate ongoing chronic GVHD via reducing activated pathogenic T and B cells.

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MicroRNA-143-3p alleviates murine collagen-induced arthritis by polarizing naive CD4+T cells into Treg cells

10.22541/au.162073400.01684976/v1 ◽

2021 ◽

Author(s):

Mei Yu Shen ◽

Bao Ping Jiang ◽

Ming Fei Zhang ◽

Xiang Wang ◽

Hong Zhu ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Therapeutic Strategy ◽

Treg Cells ◽

Systemic Autoimmune Disease ◽

Forkhead Box ◽

And Function ◽

Anti Inflammatory Cytokine

Rheumatoid arthritis (RA) is a chronic and systemic autoimmune disease with the decreasing proportion of regulatory T (Treg) cells. Previous studies have shown that microRNAs (miRNAs, miR) act as key regulators of Treg cells. In this study, we assessed the involvement of miR-143-3p on Treg cells differentiation and function in the RA progress. We reported that the expression of miR-143-3p has been negatively associated with RA disease activity, and actively correlated with anti-inflammatory cytokine IL-10, which was secreted by Treg cells. In vitro, miR-143-3p expression in the CD4+T cells contributed to the upregulation of forkhead box protein 3 (Foxp3), which was the characteristic transcription factor of Treg cells. Notably, miR-143-3p mimics treatment markedly upregulated the frequency of Treg cells in vivo, effectively prevented CIA development and significantly inhibited inflammation in mice. Altogether, we proposed that MiR-143-3p can alleviate CIA by polarizing naive CD4+T cells into Treg cells, which warrants miR-143-3p as a target for the new therapeutic strategy of Treg-deficiency autoimmune diseases such as RA.

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Comparison of In Vitro and In Vivo Effects of Infliximab on Cytokine Production in Proliferating CD4+ T Cells in Patients with Rheumatoid Arthritis

Clinical Anti-Inflammatory & Anti-Allergy Drugs ◽

10.2174/2212703802666150127001412 ◽

2015 ◽

Vol 1 (2) ◽

pp. 122-128

Author(s):

Syuichi Koarada ◽

Yuri Sadanaga ◽

Natsumi Nagao ◽

Satoko Tashiro ◽

Rie Suematsu ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

T Cells ◽

Cd4 T Cells ◽

Cytokine Production

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mTOR pathway regulates the differentiation of peripheral blood Th2/Treg cell subsets in patients with pemphigus vulgaris

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/gmab008 ◽

2021 ◽

Author(s):

Kuan Lai ◽

Wenjing Zhang ◽

Songshan Li ◽

Zhiwen Zhang ◽

Shuangde Xie ◽

...

Keyword(s):

T Cells ◽

Peripheral Blood ◽

Cd4 T Cells ◽

Pemphigus Vulgaris ◽

Treg Cells ◽

Mtor Pathway ◽

Cd4 T Cell ◽

Cell Ratio ◽

Loss Of Balance

Abstract Pemphigus vulgaris (PV) is a chronic and potentially life-threatening autoimmune blistering disease. Aberrant mTOR pathway activity is involved in many autoimmune diseases. This study investigated the correlation of mTOR pathway (PI3K/AKT/mTOR/p70S6K) activity with the loss of balance in T helper 2/regulatory T (Th2/Treg) cells in the peripheral blood of PV patients. CD4+ T cells were isolated from 15 PV patients and 15 healthy controls (HCs), the ratios of Th2/CD4+ T cells and Treg/CD4+ T cells, the activity of the mTOR pathway (PI3K/AKT/mTOR/p70S6K), the transcription factors and cytokines of Th2 and Treg cells were detected. Primary CD4+ T cells from PV patients were cultured under Th2- or Treg-polarizing conditions with or without rapamycin in vitro. We found that PV patients showed significantly elevated serum IL-4 when compared with HCs, and serum IL-4 level was positively correlated with the titer of anti-Dsg1/3 antibody and disease severity, while the serum TGF-β level was negatively correlated with the titer of anti-Dsg3 antibody and disease severity. Meanwhile, PV patients showed increased Th2/CD4+ T cell ratio; decreased Treg/CD4+ T cell ratio; elevated mRNA of PI3K, AKT, mTOR and protein of PI3K (P85), AKT, p-AKT (Ser473), mTOR, p-mTOR (Ser2448), p-p70S6K (Thr389), GATA3; reduced protein of forkhead box protein 3. Rapamycin inhibited Th2 cell differentiation and promoted Treg cell differentiation in vitro. These data suggest a close association between mTOR pathway activation and the loss of balance in Th2/Treg cells in peripheral blood of PV patients. Inhibiting mTORC1 can help restore the Th2/Treg balance.

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Cd40-Independent Pathways of T Cell Help for Priming of Cd8+ Cytotoxic T Lymphocytes

Journal of Experimental Medicine ◽

10.1084/jem.191.3.541 ◽

2000 ◽

Vol 191 (3) ◽

pp. 541-550 ◽

Cited By ~ 144

Author(s):

Zhengbin Lu ◽

Lingxian Yuan ◽

Xianzheng Zhou ◽

Eduardo Sotomayor ◽

Hyam I. Levitsky ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cd4 T Cells ◽

Cytotoxic T Lymphocyte ◽

Cell Communication ◽

Cd8 T Cell ◽

Cd4 Help ◽

T Cell Help

In many cases, induction of CD8+ CTL responses requires CD4+ T cell help. Recently, it has been shown that a dominant pathway of CD4+ help is via antigen-presenting cell (APC) activation through engagement of CD40 by CD40 ligand on CD4+ T cells. To further study this three cell interaction, we established an in vitro system using dendritic cells (DCs) as APCs and influenza hemagglutinin (HA) class I and II peptide–specific T cell antigen receptor transgenic T cells as cytotoxic T lymphocyte precursors and CD4+ T helper cells, respectively. We found that CD4+ T cells can provide potent help for DCs to activate CD8+ T cells when antigen is provided in the form of either cell lysate, recombinant protein, or synthetic peptides. Surprisingly, this help is completely independent of CD40. Moreover, CD40-independent CD4+ help can be documented in vivo. Finally, we show that CD40-independent T cell help is delivered through both sensitization of DCs and direct CD4+–CD8+ T cell communication via lymphokines. Therefore, we conclude that CD4+ help comprises at least three components: CD40-dependent DC sensitization, CD40-independent DC sensitization, and direct lymphokine-dependent CD4+–CD8+ T cell communication.

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