B‐cell‐derived IL‐10 does not vitally contribute to the clinical course of glomerulonephritis

Kluger, Malte A.; Ostmann, Annett; Luig, Michael; Meyer, M Cobas; Goerke, Boeren; Paust, Hans‐Joachim; Meyer-Schwesinger, Catherine; Stahl, Rolf A.K.; Panzer, Ulf; Tiegs, Gisa; Steinmetz, Oliver M.

doi:10.1002/eji.201343842

Cited by 5 publications

(7 citation statements)

References 51 publications

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“…These mice were then mated to another strain expressing red fluorescent protein under the Foxp3 promoter and a green fluorescent protein under control of the IL-10 promoter. 42 In these mice, all T cells, including Tregs, lack the IL-10Ra. In addition, Foxp3 + Tregs as well as their IL-10 production can be detected reliably by autofluorescence.…”

Section: Discussionmentioning

confidence: 99%

“…40 Sheep globulin-specific serum IgG titers were analyzed by ELISA (for total IgG; Biozol, Eching, Germany; for IgG1, IgG2b, IgG2c, and IgG3; Invitrogen, Frederick, MD). 13,42 Flow Cytometry Cells were surface stained for 30 minutes at 4°C with fluorochrome-labeled antibodies against CD45, CD45.1 CD45.2, CD4, CD44, CD11c, CCR7 (all BD Biosciences, Franklin Lakes, NJ), CD3, CD8, gdTCR, CD25, CD69, CD62L, CCR6, CXCR3, ICOS, PD-1, CD103, PD-L1, GITR, CTLA-4, CD39, CD73 (all Biolegend), MHCII (eBiosciences), and Granzyme B (Thermo Fischer Scientific, Waltham, MA) as previously described. 20,22 For intracellular and intranuclear staining, samples were processed using a commercial intranuclear staining kit (Foxp3 Staining Kit; eBiosciences).…”

Section: Isolation Of Leukocytes From Various Tissuesmentioning

confidence: 99%

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IL-10 Receptor Signaling Empowers Regulatory T Cells to Control Th17 Responses and Protect from GN

Diefenhardt¹,

Nosko²,

Kluger³

et al. 2018

JASN

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Th17 cells are central pathogenic mediators of autoimmune disease, including many forms of GN. IL-10 receptor signaling (IL-10R) in regulatory T cells (Tregs) has been implicated in the downregulation of Th17 cells, but the underlying molecular mechanisms and functional relevance of this process remain unclear. We generated mice with Treg-specific IL-10Ra deficiency and subjected these mice to nephrotoxic serum-induced nephritis as a model of crescentic GN. Immune responses and Treg phenotypes were extensively analyzed. Compared with controls, mice with IL-10Ra Tregs showed a spontaneously overshooting Th17 immune response. This hyper-Th17 phenotype was further boosted during GN and associated with aggravated renal injury. Notably, abrogation of IL-10Ra signaling in Tregs increased dendritic cell activation and production of Th17-inducing cytokines. In contrast, Treg trafficking and expression of chemokine receptor CCR6 remained unaffected, indicating mechanisms of Th17 control, differing from those of previously identified CCR6 Treg17 cells. Indeed, the capacity for direct suppression of Th17 responses by IL-10Ra Tregs was significantly impaired. As underlying pathology, analyses conducted and using double-fluorescent reporter mice revealed strikingly decreased IL-10 production by IL-10Ra Tregs. To assess, whether reduced IL-10 could explain the hyper Th17 phenotype, competitive cotransfer experiments were performed. Supporting our concept, IL-10Ra T cells differentiated into Th17 cells at much higher frequencies than wild type T cells did during GN. IL-10R engagement optimizes Treg-mediated suppression of Th17 immunity. We hypothesize a feed-forward loop, in which IL-10Ra signaling reinforces IL-10 secretion by Tregs which potently controls Th17 development direct and indirect mechanisms. IL-10R thus may be a promising therapeutic target for the treatment of GN.

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Section: Discussionmentioning

confidence: 99%

Section: Isolation Of Leukocytes From Various Tissuesmentioning

confidence: 99%

IL-10 Receptor Signaling Empowers Regulatory T Cells to Control Th17 Responses and Protect from GN

Diefenhardt¹,

Nosko²,

Kluger³

et al. 2018

JASN

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“…20 Paraffin-embedded sections were stained with antibodies directed against murine fibrin alpha chain (UC45; Abcam, Inc., Cambridge, UK), CD3 (A0452; Dako, Hamburg, Germany), F4/80 (BM8; BMA Biomedicals, Hiddenhausen, Germany), MAC2 (M3/38; Cedarlane-Laboratories, Burlington, ON, Canada), GR-1 or Foxp3 and developed with a polymer-based secondary antibody-alkaline phosphatase kit (POLAP; Zytomed, Berlin, Germany), as published previously. 40 Fifty glomerular cross-sections (gcs) and 30 tubulointerstitial high power fields (hpf, magnification, 3400) per kidney section were counted in a blinded fashion.…”

Section: Morphologic Studiesmentioning

confidence: 99%

RORγt+Foxp3+ Cells are an Independent Bifunctional Regulatory T Cell Lineage and Mediate Crescentic GN

Kluger

Meyer

Nosko

et al. 2016

Journal of the American Society of Nephrology

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Cells expressing both the regulatory T cell (Treg)-inducing transcription factor Foxp3 and the Th17 transcription factor RORgt have been identified (biTregs). It is unclear whether RORgt + Foxp3 + biTregs belong to the Th17-specific Treg17 cells, represent intermediates during Treg/Th17 transdifferentiation, or constitute a distinct cell lineage. Because the role of biTregs in inflammatory renal disease is also unknown, we studied these cells in the nephrotoxic nephritis (NTN) model of acute crescentic GN. Induction of NTN resulted in rapid renal and systemic expansion of biTregs. Notably, analyses of the biTreg expression profile revealed production of both anti-inflammatory (IL-10, IL-35) and proinflammatory (IL-17) cytokines. Additionally, biTregs expressed a signature of surface molecules and transcription factors distinct from those of Th17 cells and conventional Tregs (cTregs), and biTregs were identified in Treg17-deficient mice. Finally, fate reporter and cell transfer studies confirmed that biTregs are not Treg/Th17 transdifferentiating cells. Therapeutic transfer of biTregs suppressed the development of nephritis to an extent similar to that observed with transferred cTregs, but in vitro studies indicated different mechanisms of immunosuppression for biTregs and cTregs. Intriguingely, as predicted from their cytokine profile, endogenous biTregs displayed additional proinflammatory functions in NTN that were abrogated by cellspecific deletion of RORgt. In summary, we provide evidence that RORgt + Foxp3 + biTregs are a novel and independent bifunctional regulatory T cell lineage distinct from cTregs, Treg17 cells, and Th17 cells. Furthermore, biTregs appear to contribute to crescentic GN and hence may be novel therapeutic targets.

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“…Fifty glomerular cross-sections and 30 tubulointerstitial high power fields (magnification of 3400) per kidney section were counted in a blinded fashion. 13,41…”

Section: Morphologic Studiesmentioning

confidence: 99%

“…40 Circulating sheep globulin-specific serum IgG titers were analyzed by ELISA using plates precoated with sIgG. 13,41 For analysis of total nonantigen-specific Igs, ELISA plates were precoated with anti-mouse total IgG antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA). ELISA plates were incubated with mouse serum, and the following secondary antibodies were used for detection: total IgG (Southern Biotech, Birmingham, AL), IgG1 (Southern Biotech), IgG2b (Invitrogen, Carlsbad, CA), IgG2c (Bethyl, Montgomery, TX), and IgG3 (Jackson ImmunoResearch Laboratories).…”

Section: Systemic Cellular and Humoral Immune Responsesmentioning

confidence: 99%

T-Bet Enhances Regulatory T Cell Fitness and Directs Control of Th1 Responses in Crescentic GN

Nosko¹,

Kluger²,

Diefenhardt³

et al. 2016

JASN

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Th1 cells are central pathogenic mediators of crescentic GN (cGN). Mechanisms responsible for Th1 cell downregulation, however, remain widely unknown. Recently, it was proposed that activation of the Th1-characteristic transcription factor T-bet optimizes Foxp3 regulatory T (Treg) cells to counteract Th1-type inflammation. Because very little is known about the role of T-bet Treg1 cells in inflammatory diseases, we studied the function of these cells in the nephrotoxic nephritis (NTN) model of cGN. The percentage of Treg1 cells progressively increased in kidneys of nephritic wild-type mice during the course of NTN, indicating their functional importance. Notably, naïve Foxp3xT-bet mice, lacking Treg1 cells, showed spontaneous skewing toward Th1 immunity. Furthermore, absence of Treg1 cells resulted in aggravated NTN with selectively dysregulated renal and systemic Th1 responses. Detailed analyses of Treg cells from Foxp3xT-bet mice revealed unaltered cytokine production and suppressive capacity. However, in competitive cotransfer experiments, wild-type Treg cells outcompeted T-bet-deficient Treg cells in terms of population expansion and expression levels of Foxp3, indicating that T-bet expression is crucial for general Treg fitness. Additionally, T-bet-deficient Treg cells lacked expression of the Th1-characteristic trafficking receptor CXCR3, which correlated with significant impairment of renal Treg infiltration. In summary, our data indicate a new subtype of Treg cells in cGN. These Treg1 cells are characterized by activation of the transcription factor T-bet, which enhances the overall fitness of these cells and optimizes their capacity to downregulate Th1 responses by inducing chemokine receptor CXCR3 expression.

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B‐cell‐derived IL‐10 does not vitally contribute to the clinical course of glomerulonephritis

Cited by 5 publications

References 51 publications

IL-10 Receptor Signaling Empowers Regulatory T Cells to Control Th17 Responses and Protect from GN

IL-10 Receptor Signaling Empowers Regulatory T Cells to Control Th17 Responses and Protect from GN

RORγt+Foxp3+ Cells are an Independent Bifunctional Regulatory T Cell Lineage and Mediate Crescentic GN

T-Bet Enhances Regulatory T Cell Fitness and Directs Control of Th1 Responses in Crescentic GN

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