TNF Receptor Type I-Dependent Activation of Innate Responses to Reduce Intestinal Damage-Associated Mortality

doi:10.1053/j.gastro.2007.11.055

Gastroenterology

Volume 134, Issue 2, February 2008, Pages 470-480

https://doi.org/10.1053/j.gastro.2007.11.055 Get rights and content

Background & Aims: Ligation of tumor necrosis factor (TNF) receptors (TNFRs) with TNF plays a critical role in the pathogenesis of human inflammatory bowel disease (IBD). However, it remains unclear which cell types activated through TNFR-associated signaling cascades are involved in the pathogenesis of colitis. Methods: Recombination activating gene-1 (RAG) knockout (KO) (no T or B cells)-based TNFR double and triple KO mice were generated. Bone marrow (BM) chimera mice in which BM-derived myeloid cells, but not colonic epithelial cells (CECs), express TNFRs were also generated. Colitis was induced by administration of dextran sodium sulfate (DSS) in distilled water. Murine lines and chimeras were assessed for disease severity, histopathology, apoptotic cell rate, epithelial proliferation, and bacterial invasion rate. Results: Following DSS administration, mice lacking both RAG and TNFR1 exhibited a high mortality (>80%) rate with an impaired CEC regeneration compared with RAG KO and RAG × TNFR2 double KO (DKO) mice. Transplantation of RAG KO-derived BM cells restored CEC regeneration and rescued the majority of recipient RAG × TNFR1 DKO mice from DSS-induced mortality. After BM transplantation, RAG × TNFR1 DKO mice exhibited an increased rate of apoptosis in the colonic lamina propria macrophages in association with the activation of caspases. In addition, BM reconstitution directly or indirectly enhanced the proliferation of CECs by activating mitogen-activated protein kinase and phosphoinositide-3 kinase/Akt pathways. Conclusions: TNFR1-signaling cascade in colonic myeloid lineage cells contributes to the suppression of acute damage-associated mortality presumably by controlling CEC homeostasis.

Section snippets

Mice and Bone Marrow Transplantation

Six to 8-week-old wild-type (WT), recombination activating gene-1 (RAG) knockout (KO), TNFR1 KO, and TNFR2 KO mice on the C57BL/6 strain (H-2^b) were purchased from the Jackson Laboratories (Bar Harbor, ME). To generate RAG KO mice deficient in TNFR1 or TNFR2, RAG KO mice were crossed with TNFR1 KO or TNFR2 KO mice. DNA screening was performed as previously described.¹³ Briefly, genomic DNA extracted from tail was subject to polymerase chain reaction (PCR). To distinguish heterozygous and

Distinct Effects of TNFR1 vs TNFR2 on Innate Immune Responses Under Acute Intestinal Damage

TNF/TNFR2-mediated signaling on LP T cells is known to play a critical role in the pathogenesis of CD and experimental colitis.19, 20, 21, 22 However, the role of TNFRs in innate immune responses involved in the pathogenesis of colitis has not fully been defined. To address this question, we generated TNFR1 or TNFR2-deficient RAG double KO (DKO) mice that lack T or B cells and induced acute intestinal inflammation by the administration of 4% DSS for 5 days (Figure 1A). After cessation of DSS

Discussion

Using a model of acute intestinal inflammation in which acquired immune responses can be abrogated, we here demonstrate that the pathophysiologic roles of TNFR1 and TNFR2 in innate immune pathways differ during acute intestinal inflammation. As observed in acquired immune responses,²⁰ TNFR2-mediated activation of innate immune cells in the colonic LP contributes to the exacerbation of colitis. In contrast, TNFR1 expressing LP cells are required for epithelial regeneration following acute

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However, the therapeutic effects of anti-TNF agents are short lived for many (Ben-Horin et al., 2014; Colombel et al., 2007; Ford et al., 2011; Schreiber et al., 2007). In mice, deletion of TNF or either of its transmembrane receptors, TNFR1 (p55) and TNFR2 (p75) (encoded by the Tnfrsf1a and Tnfrsf1b genes, respectively), has differing effects on disease susceptibility in established colitis models (Dubé et al., 2015; Ebach et al., 2005; Mizoguchi et al., 2008; Wang et al., 2012; Wang et al., 2013). Strikingly, loss of TNF-signaling-pathway members Tnf (Hale and Greer, 2012) or Tnfr2 (Punit et al., 2015) results in severe disease in the Il10−/− spontaneous colitis model (Kühn et al., 1993).
Neutralization of tumor necrosis factor (TNF) represents a widely used therapeutic strategy for autoimmune diseases including inflammatory bowel disease (IBD). However, the fact that many patients with IBD are non-responsive to anti-TNF therapies suggests the need for a better understanding of TNF signaling in IBD. Here, we show that co-deletion of TNF receptor 1 (TNFR1, Tnfrsf1a) in the Il10^−/− spontaneous colitis model exacerbates disease, resulting in very-early-onset inflammation after weaning. The disease can be interrupted by treatment with antibiotics. The single deletion of TNFR1 induces subclinical colonic epithelial dysfunction and mucosal immune abnormalities, including accumulation of neutrophils and depletion of B cells. During the pre-disease period (before weaning), both Tnfr1^−/− and Il10^−/− Tnfr1^−/− animals exhibit impaired expression of pro-inflammatory cytokines compared with wild-type and Il10^−/− controls, respectively. Collectively, these results demonstrate the net anti-inflammatory functions of TNF/TNFR1 signaling through the regulation of colonic immune homeostasis in early life.
Lysophosphatidic Acid-Mediated GPR35 Signaling in CX3CR1<sup>+</sup> Macrophages Regulates Intestinal Homeostasis
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Single-nucleotide polymorphisms in the gene encoding G protein-coupled receptor 35 (GPR35) are associated with increased risk of inflammatory bowel disease. However, the mechanisms by which GPR35 modulates intestinal immune homeostasis remain undefined. Here, integrating zebrafish and mouse experimental models, we demonstrate that intestinal Gpr35 expression is microbiota dependent and enhanced upon inflammation. Moreover, murine GPR35⁺ colonic macrophages are characterized by enhanced production of pro-inflammatory cytokines. We identify lysophosphatidic acid (LPA) as a potential endogenous ligand produced during intestinal inflammation, acting through GPR35 to induce tumor necrosis factor (Tnf) expression in macrophages. Mice lacking Gpr35 in CX3CR1⁺ macrophages aggravate colitis when exposed to dextran sodium sulfate, which is associated with decreased transcript levels of the corticosterone-generating gene Cyp11b1 and macrophage-derived Tnf. Administration of TNF in these mice restores Cyp11b1 expression and intestinal corticosterone production and ameliorates DSS-induced colitis. Our findings indicate that LPA signals through GPR35 in CX3CR1⁺ macrophages to maintain TNF-mediated intestinal homeostasis.
Interleukin 32, inflammation and cancer
2017, Pharmacology and Therapeutics
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Severity levels of DSS (dextran sodium sulfate)-induced colitis were lower in IL-32γ TG mice, compared with colitis in the wild type mice (Choi et al., 2010; Oh et al., 2011). Unexpectedly these results are similar in that DSS-induced colitis was acerbated in deficient mice of TNFα or TNFR1 compared to the wild type mice (Mizoguchi et al., 2008; Naito et al., 2003; Noti, Corazza, Mueller, Berger, & Brunner, 2010). These results suggest that IL-32 has a dual function in innate responses of the intestine, which constitutive expression of TNFα and IL-10 by IL-32 may regulate expression of inflammatory cytokine by NOD2 and protect the barrier from pathogens.
Interleukin-32 (IL-32) is a novel cytokine involved in inflammation and cancer development. IL-32 gene consists of eight small exons, and IL-32 mRNA has nine alternative spliced isoforms, and was thought to be secreted because it contains an internal signal sequence and lacks a transmembrane region. IL-32 is initially expressed selectively in activated T cells by mitogen and activated NK cells and their expression is strongly augmented by microbes, mitogens, and other cytokines. The IL-32 is induced mainly by pathogens and pro-inflammatory cytokines, but IL-32 is more prominent in immune cells than in non-immune tissues. The IL-32 transcript is expressed in various human tissues and organs such as the spleen, thymus, leukocyte, lung, small intestine, colon, prostate, heart, placenta, liver, muscle, kidney, pancreas, and brain. Cytokines are critical components of cell signaling pathways that are involved in the regulation of cell growth, metabolism, hormone signaling, immune regulation and a variety of other physiological functions. Earlier studies have demonstrated that IL-32 regulates cell growth, metabolism and immune regulation and is therefore involved in the pathologic regulator or protectant of inflammatory diseases. Previous studies defined that IL-32 is upregulated in the patients with several inflammatory diseases, and is induced by inflammatory responses. However, several reports suggested that IL-32 is downregulated in several inflammatory diseases including asthma, HIV infection disease, neuronal diseases, metabolic disorders, experimental colitis and metabolic disorders. IL-32 is also involved in various cancer malignancies including renal cancer, esophageal cancer and hepatocellular carcinoma, lung cancer, gastric cancer, breast cancer, pancreatic cancer, lymphoma, osteosarcoma, breast cancer, colon cancer and thyroid carcinoma. Other studies suggested that IL-32 decreases tumor development including cervical cancer, colon cancer and prostate cancer, melanoma, pancreatic cancer, liver cancer and chronic myeloid leukemia. Nevertheless, review articles that discuss the roles and its mechanism of IL-32 isoforms focusing on the therapeutic approaches have not yet been reported. In this review article, we will discuss recent findings regarding IL-32 in the development of diseases and further discuss therapeutic approaches targeting IL-32. Moreover, we will suggest that IL-32 could be the target of several diseases and the therapeutic agents for targeting IL-32 may have potential beneficial effects for the treatment of inflammatory diseases and cancers. Future research should open new avenues for the design of novel therapeutic approaches targeting IL-32.
Tumor Necrosis Factor Receptor 2 Restricts the Pathogenicity of CD8<sup>+</sup> T Cells in Mice With Colitis
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Tumor necrosis factor receptor 2 (TNFR2, Tnfrsf1b) regulates multiple aspects of immune function, but little is known about its role in the immunopathogenesis of inflammatory bowel disease (IBD). We investigated whether TNFR2 restricts the activity of specific immune cell subtypes to protect against the development of colitis in mice.
Tnfr2^−/− mice were crossed with interleukin (Il) 10^−/− mice, which spontaneously develop colitis, to generate Il10^−/−Tnfr2^−/− mice. Colonic tissues were collected from Il10^−/−Tnfr2^−/− mice along with Il10^−/− mice (controls) and analyzed by flow cytometry and histology. Bone marrow was transplanted into Il10^−/− and Il10^−/−Tnfr2^−/− mice from Il10^−/− or Il10^−/−Tnfr2^−/− donors by intravenous injection. CD8⁺ T cells were neutralized in Il10^−/−Tnfr2^−/− mice by intraperitoneal injection of anti-CD8 or isotype control antibodies. Colitis was induced in Rag2^−/− mice by intravenous injections of naïve CD8⁺ T cells isolated from C57BL/6 or Tnfr2^−/− mice.
Il10^−/−Tnfr2^−/− mice spontaneously developed more severe colitis compared with Il10^−/− controls, characterized by selective expansion of colonic CD8⁺ T cells. Transplantation of TNFR2-deficient bone marrow resulted in significantly increased incidence and severity of colitis. Transcriptome analyses showed that the expression of genes regulated by TNFR2 were specific to CD8⁺ T cells and included genes associated with risk for IBD. Depletion of CD8⁺ T cells from Il10^−/−Tnfr2^−/− mice prevented colonic inflammation. Adoptive transfer of TNFR2-null naïve CD8⁺ T cells compared with CD8⁺ T cells from control mice increased the severity of colitis that developed in Rag2^−/− mice.
TNFR2 protects mice from colitis by inhibiting the expansion of colonic CD8⁺ T cells. TNFR2 regulates expression of genes that regulate CD8⁺ T cells and have been associated with susceptibility to IBD. Disruption in TNFR2 signaling might therefore be associated with pathogenesis. Strategies to increase levels or activity of TNFR2 and thereby reduce the activity of CD8⁺ T cells might be developed to treat IBD patients with CD8⁺ T cell dysfunction.
TNFR2 activates mlck-dependent tight junction dysregulation to cause apoptosis-mediated barrier loss and experimental colitis
2013, Gastroenterology
Citation Excerpt :
This conclusion, which is consistent with in vitro data showing that TNFR2 signaling mediates TNF-induced long MLCK expression and tight junction regulation,6 might explain the association of TNFR2 polymorphisms with Crohn’s disease.35 Although a protective effect of TNFR2 knockout has recently been reported in DSS colitis,36 this result is controversial, as other studies have demonstrated no significant change or increased DSS sensitivity in TNFR2 knockout mice.37,38 One possible explanation for this disparity between studies is that differences in mouse strain used, local microbiome, or other factors that affect DSS colitis development highlighted or suppressed the well known role of TNFR2 in promoting epithelial migration and wound healing.8
Tight junction dysregulation and epithelial damage contribute to barrier loss in patients with inflammatory bowel disease. However, the mechanisms that regulate these processes and their relative contributions to disease pathogenesis are not completely understood. We investigated these processes using colitis models in mice.
We induced colitis by adoptive transfer of CD4⁺CD45RB^hi cells or administration of dextran sulfate sodium to mice, including those deficient in tumor necrosis factor receptor (TNFR) 1, TNFR2, or the long isoform of myosin light chain kinase (MLCK). Intestinal tissues and isolated epithelial cells were analyzed by immunoblot, immunofluorescence, enzyme-linked immunosorbent assay, and real-time polymerase chain reaction assays.
Induction of immune-mediated colitis by CD4⁺CD45RB^hi adoptive transfer increased intestinal permeability, epithelial expression of claudin-2, the long isoform of MLCK, and TNFR2 (but not TNFR1) and phosphorylation of the myosin II light chain. Long MLCK upregulation, myosin II light chain phosphorylation, barrier loss, and weight loss were attenuated in TNFR2^−/− , but not TNFR1^−/− , recipients of wild-type CD4⁺CD45RB^hi cells. Similarly, long MLCK^−/− mice had limited increases in myosin II light chain phosphorylation, claudin-2 expression, and intestinal permeability and delayed onset of adoptive transfer−induced colitis. However, coincident with onset of epithelial apoptosis, long MLCK^−/− mice ultimately developed colitis. This indicates that disease progresses via apoptosis in the absence of MLCK-dependent tight junction regulation. In support of this conclusion, long MLCK^−/− mice were not protected from epithelial apoptosis-mediated, damage-dependent dextran sulfate sodium colitis.
In immune-mediated inflammatory bowel disease models, TNFR2 signaling increases long MLCK expression, resulting in tight junction dysregulation, barrier loss, and induction of colitis. At advanced stages, colitis progresses by apoptosis and mucosal damage that result in tight junction- and MLCK-independent barrier loss. Therefore, barrier loss in immune-mediated colitis occurs via two temporally and morphologically distinct mechanisms. Differential targeting of these mechanisms can lead to improved inflammatory bowel disease therapies.
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2013, Animal Models for the Study of Human Disease
Inflammatory bowel disease (IBD) is a chronic, relapsing intestinal inflammatory condition with increasing incidence and prevalence around the world. Conventional and biological therapies have been successfully, or unsuccessfully, applied for the treatment of IBD and several new biological therapies are currently in clinical development. So far, more than 69 kinds of animal models have been established to study IBD, and they have provided significant contributions not only for understanding the mechanism of IBD but also for developing the rationale of new therapies. Although individual animal models can not fully reflect the pathogenesis of human IBD, the data from animal models—if a consensus is reached from many but not from one or few—may be used as a predictive factor of effectiveness for some biological therapies. Therefore, in this Chapter, we will discuss this possibility by comparing the data from animal models of IBD with data from some clinical trials.

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: Conflicts of interest: None of the authors have conflicts of interest to disclose.

: Supported by NIH grants DK64289, DK74454 (to E.M.), DK43351, DK60049 (to D.K.P.), and DK 64351 (to A.M.), and IBD Grants from Broad Medical Foundation (to E.M.).

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Basic–Alimentary TractTNF Receptor Type I-Dependent Activation of Innate Responses to Reduce Intestinal Damage-Associated Mortality

Section snippets

Mice and Bone Marrow Transplantation

Distinct Effects of TNFR1 vs TNFR2 on Innate Immune Responses Under Acute Intestinal Damage

Discussion

Gastroenterology

Gastroenterology

Gastroenterology

Exp Cell Res

Gastroenterology

Gastroenterology

Trends Immunol

Gastroenterology

Gastroenterology

Cell

Inflammatory bowel disease

N Eng J Med

The immunology of mucosal models of inflammation

Annu Rev Immunol

Immune networks in animal models of inflammatory bowel disease

Inflamm Bowel Dis

Role of cytokines in the early stages of chronic colitis in TCRα-mutant mice

Lab Invest

Evidence for function of Ia molecules on gut epithelial cells in man

J Exp Med

Tumor necrosis factor receptor and Fas signaling mechanisms

Annu Rev Immunol

TNF-R1 signaling: a beautiful pathway

Science

Basic–Alimentary Tract
TNF Receptor Type I-Dependent Activation of Innate Responses to Reduce Intestinal Damage-Associated Mortality