Thalidomide attenuates excessive inflammation without interrupting lipopolysaccharide-driven inflammatory cytokine production in chronic granulomatous disease

doi:10.1016/j.clim.2013.03.004

Clinical Immunology

Volume 147, Issue 2, May 2013, Pages 122-128

https://doi.org/10.1016/j.clim.2013.03.004 Get rights and content

Highlights

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Thalidomide suppressed TNFα-induced NF-κB activation in CGD monocytes.
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Thalidomide did not interrupt LPS-driven cytokine production in CGD monocytes.
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CGD monocytes released higher levels of IL-6 and TNF-α in response to LPS.
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Thalidomide reduced serum level of inflammatory cytokines in CGD colitis patient.
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Thalidomide did not impact immunity to pathogenic infections in CGD patient.

Abstract

Chronic granulomatous disease (CGD) is a rare inherited disorder characterized by an inability to produce reactive oxygen species, resulting in recurrent life-threatening infections. Curiously, half of the patients with CGD suffer from aseptic bowel inflammation (CGD colitis) due to dysregulated inflammation induced by TNF-α and IL-1β. Thus, developing therapies that regulate excessive inflammatory responses without interrupting antimicrobial immunity would benefit CGD colitis patients. Here, we show that thalidomide suppressed TNF-α-induced NF-κB activation and ATP-induced IL-1β secretion, but did not interrupt the production of IL-1β, IL-6, IL-8, and TNF-α in response to lipopolysaccharide in CGD monocytes. We report on a CGD colitis patient that showed decreased bowel inflammation characterized by reduced serum levels of inflammatory cytokines without evidence of progression of fungal and bacterial infections present at initiation of thalidomide therapy. Our results suggest that thalidomide could be an efficacious therapeutic option for patients with CGD colitis suffering from serious infections.

Introduction

Reactive oxygen species (ROS) generated by cellular nicotinamide adenine dinucleotide phosphate (NADPH) oxidase play an important role in killing microorganisms ingested by phagocytes. In addition, ROS function as pleiotropic signals in various inflammatory contexts. For example, ROS activate nuclear factor-κB (NF-κB), which produces inflammatory cytokines and facilitates formation of the inflammasome consisting of Nod-like receptor family, pyrin domain containing 3 (NLRP3), ASC, cardinal, and pro-caspase-1, which cleaves into caspase-1 and converts pro-interleukin-1β (IL-1β) to IL-1β [1], [2]. Chronic granulomatous disease (CGD) is a primary immunodeficiency resulting from the inability to produce ROS due to defects in NADPH oxidase activity [3]. CGD patients suffer from recurrent infection and frequently develop CGD-associated bowel inflammation (CGD colitis) in the absence of any demonstrable infection [4]. The underlying mechanism of CGD colitis is not fully understood; however, dysregulated inflammatory cytokine production, such as IL-1β and tumor necrosis factor-α (TNF-α), by monocytes and macrophages is thought to contribute to the disease [1], [5]. This concept is supported by many clinical reports that neutralizing anti-TNF-α or immunosuppressive drugs, such as steroids or azathioprine, effectively mitigate CGD colitis clinical symptoms. Notably, the use of these drugs results in an increased susceptibility to severe infections [4], [5].

Thalidomide, originally developed in the 1950s as a tranquilizer, was discontinued when its potent teratogenic effects resulted in catastrophic birth defects in the 1960s. It is recently being reconsidered for its unique and pleiotropic medical effects, such as immunomodulation and anti-inflammatory properties. Thalidomide potentially reduces inflammation by blocking nuclear localization of NF-κB and inhibiting inflammatory cytokine production in patients with chronic inflammatory diseases such as Behcet's disease, rheumatoid arthritis, and Crohn disease [4], [6], [7], [8]. So far, there are case reports regarding three patients with CGD colitis who were treated with thalidomide [4], [9], although the immunomodulatory effect of thalidomide on CGD monocytes has not been described. In this report we show that thalidomide suppressed TNF-α-induced NF-κB activation and adenosine triphosphate (ATP)-induced IL-1β secretion while not interrupting lipopolysaccharide (LPS)-driven inflammatory cytokine production in CGD monocytes. We also show that thalidomide treatment improved bowel inflammation by reducing serum TNF-α levels without increasing susceptibility to infection in a CGD colitis patient.

Section snippets

Monocyte isolation and stimulation

All experiments were performed following receipt of informed consent from patients with X-linked CGD or their parents as part of a protocol approved by the Institutional Review Board of the National Center for Child Health and Development. Monocytes isolated from peripheral blood by an antibody-mediated cell enrichment procedure from RossettSep (StemCell Technologies, Canada) were stimulated with serial concentrations of LPS and with carbonyl cyanide 3-chlorophenylhydrazone (CCCP)

Thalidomide does not block caspase-1 activation or IL-1β production following LPS stimulation in CGD monocytes

Monocytes are capable of activating caspase-1 to process and release IL-1β by LPS signaling through Toll-like receptor 4 (TLR4) [11]. In keeping with these results [12], LPS induced capase-1 activation and IL-1β production in a dose-dependent manner in CGD monocytes as well as in healthy control monocytes (Figs. 1a–c), suggesting that ROS generated by NADPH oxidase was dispensable for caspase-1 activation and conversion of pro-IL-1β to IL-1β. To assess the effect of thalidomide on caspase-1

Discussion

The paradoxical clinical manifestations of CGD, susceptibility to infection and excessive inflammation, make it difficult to choose an appropriate course of therapy for patients with CGD colitis that develops through dysregulated inflammatory cytokine production [19], [20], [21]. The reason for this may be attributed to our insufficient understanding of a role for ROS on inflammasome activation. CGD patient monocytes were predisposed to secrete IL-1β in response to LPS, while CGD patients

Conflict of interest

The authors declare that they have no conflicts of interest.

Acknowledgments

We are grateful to the CGD patients and blood donors for their participation in this study. This work was supported by a Grant-in-Aid for Young Scientists (B) and a Grant from the National Center for Child Health and Development.

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Pulmonary complications of Chronic Granulomatous Disease
2024, Revue des Maladies Respiratoires
La granulomatose septique chronique (CGD) est un déficit immunitaire héréditaire qui affecte les capacités oxydatives des cellules phagocytaires et se caractérise par une susceptibilité accrue aux infections bactériennes et fongiques ainsi que par une hyper inflammation granulomateuse. Les manifestations pulmonaires, fréquentes, sont de 2 types : (1) infectieuses : souvent très sévères, découvertes à un stade tardif du fait du caractère pauci-symptomatique initial. Leur incidence a diminué grâce aux prophylaxies anti-infectieuses systématiques mais reste une cause majeure de morbi-mortalité ; (2) inflammatoires : masse granulomateuse persistante ou pneumopathie interstitielle. La récurrence des complications pulmonaires depuis l’enfance génère des séquelles parenchymateuses et bronchiques impactant le pronostic fonctionnel. L’allogreffe de cellules souches hématopoïétiques est un traitement curatif ; sa place est discutée pour préserver le pronostic vital et limiter la morbidité de la maladie. Grâce aux progrès de prise en charge, l’espérance de vie a considérablement augmenté. De nouvelles problématiques apparaissent chez les sujets adultes : difficultés d’observance, augmentation des manifestations inflammatoires, résistances acquises aux anti-infectieux. Ceci souligne l’importance de la période de transition et la nécessité d’une prise en charge multidisciplinaire.
Chronic Granulomatosis Disease (CGD) is an inherited immune deficiency due to a mutation in the genes coding for the subunits of the NADPH oxidase enzyme that affects the oxidative capacity of phagocytic cells. It is characterized by increased susceptibility to bacterial and fungal infections, particularly Aspergillus, as well as complications associated with hyperinflammation and granulomatous tissue infiltration. There exist two types of frequently encountered pulmonary manifestations: (1) due to their being initially pauci-symptomatic, possibly life-threatening infectious complications are often discovered at a late stage. Though their incidence has decreased through systematic anti-bacterial and anti-fungal prophylaxis, they remain a major cause of morbidity and mortality; (2) inflammatory complications consist in persistent granulomatous mass or interstitial pneumoniae, eventually requiring immunosuppressive treatment. Pulmonary complications recurring since infancy generate parenchymal and bronchial sequelae that impact functional prognosis. Hematopoietic stem cell allograft is a curative treatment; it is arguably life-sustaining and may limit the morbidity of the disease. As a result of improved pediatric management, life expectancy has increased dramatically. That said, new challenges have appeared with regard to adults: difficulties of compliance, increased inflammatory manifestations, acquired resistance to anti-infectious therapies. These different developments underscore the importance of the transition period and the need for multidisciplinary management.
Gastrointestinal Manifestations of Immunodeficiency
2020, Pediatric Gastrointestinal and Liver Disease, Sixth Edition
Infants and children with primary and secondary immune deficiency often present with gastrointestinal symptoms, including diarrhea, abdominal pain, failure to thrive, intestinal infection, malabsorption, and intestinal inflammation. In addition, certain immune deficiencies predispose patients to gastrointestinal and hepatic malignancy. This chapter reviews the development of the mucosal immune system, the pathogenesis of the immune response and intestinal inflammation, and the gastrointestinal manifestations of immune deficiency. In addition, monogenic defects that result in very early onset inflammatory bowel disease are discussed in detail.
Mammalian target of rapamycin inhibition counterbalances the inflammatory status of immune cells in patients with chronic granulomatous disease
2017, Journal of Allergy and Clinical Immunology
Citation Excerpt :
Taken together, all these data suggest that rapamycin could be a useful therapy for managing inflammatory manifestations in patients with CGD. Until recently, treatment of inflammatory/granulomatous manifestations has been mainly based on corticosteroids, but steady progress in the understanding of CGD pathophysiology have prompted the use of alternative therapies, such as anti–TNF-α mAbs, thalidomide, or anakinra.4,6,7,31,32 Compared with these therapies, rapamycin has the advantage of (1) inhibiting the constitutive inflammasome activation of phagocytes from patients with CGD through autophagy induction, (2) having additional immunomodulatory properties independent of caspase-1 inhibition (ie, inhibition of secretion of TNF-α, IL-6, and IL-23 by mononuclear phagocytes and inhibition of TNF-α, IL-6, and IL-17A by T cells), and (3) being ironically an antifungal agent despite its known immunosuppressive properties.8
Chronic granulomatous disease (CGD) is a primary immunodeficiency caused by defective production of reactive oxygen species in phagocytic cells that results in life-threatening infections and severe inflammatory manifestations. The treatment of inflammatory manifestations remains challenging because it can be associated with an increased risk of infections. Previous studies have shown that phagocytes from patients with CGD display a defect in autophagy and a reactive oxygen species–independent activation of the inflammasome.
Because the intersections between autophagy and the inflammasome have been observed in patients with various diseases and microbial infections, we investigated the possible benefit of restoring the autophagy defect through rapamycin, a potent autophagy inducer, in the setting of CGD.
We studied 15 patients given a diagnosis of CGD and followed in our institution. All patients were free of any active infection at the time of the study.
We show that patients with CGD present a consistent inflammatory phenotype defined by (1) increased nonclassical and intermediate monocytes, (2) a proinflammatory state of mononuclear phagocytes with increased IL-1β and TNF-α content, (3) a T_H17 bias of CD4⁺ T cells, (4) and an increase in IL-17A–secreting neutrophil numbers. We document the reversion of CGD inflammatory status by the mammalian target of rapamycin inhibitor rapamycin on the different immune cell subsets. We also provide evidence for the enhancement of rapamycin's inhibitory effect on IL-1β secretion by the IL-1 receptor antagonist anakinra in phagocytes of patients with CGD.
Altogether, these data open new therapeutic approaches for CGD-related inflammatory manifestations.
A new pharmacological role for thalidomide: Attenuation of morphine-induced tolerance in rats
2016, Acta Anaesthesiologica Taiwanica
Citation Excerpt :
Moreover, thalidomide has potent immunomodulatory and anti-TNF-α properties. It inhibits the chemotaxis and cytokine-induced nuclear factor kappa B transcription factor gene expression.39–42 Our results also showed that thalidomide prevented the shifting of the dose–response curve and the ED50 to the right, the phenomenon occurring in morphine-tolerant group.
Tolerance to the analgesic effect is the main side effect of chronic administration of opioids. Several drugs have been studied to try to find agents to prevent the development of this phenomenon. In the present study we aimed to evaluate the effect of thalidomide on morphine-induced tolerance to the analgesic effect.
Groups of male rats were randomly rendered and received daily morphine in combination with thalidomide vehicle or thalidomide (2.5 mg/kg, 5 mg/kg, or 10 mg/kg, intraperitoneally). Nociception was measured using the plantar test apparatus. Latency time was recorded when the animal reacted to the light stimulus; licking or raising its hind paw. Treatments and evaluations continued until completion of tolerance to the analgesic effect of morphine.
Our findings indicated that tolerance was achieved following 11 days of morphine administration, while thalidomide postponed the day of tolerance completion for 4 days (2.5 mg/kg and 5 mg/kg thalidomide) or 10 days (10 mg/kg thalidomide). Moreover, thalidomide prevented the morphine-induced shift to the right of the ED₅₀ in the dose–response curve.
It was concluded that thalidomide attenuated the morphine-induced tolerance to the analgesic effect.
Thalidomide reduces mechanical hyperalgesia and depressive-like behavior induced by peripheral nerve crush in mice
2015, Neuroscience
Citation Excerpt :
It has been demonstrated that thalidomide given orally for 21 days, significantly reduced plasma levels of pro-inflammatory cytokines, such as TNF-α in streptozotocin-induced diabetes in mice (Amirshahrokhi and Ghazi-Khansari, 2012). Also, it is well known that thalidomide was able to prevent TNF-α synthesis following human monocyte stimulation by lipopolysaccharides (LPS) (Kawai et al., 2013), and it was able to prevent the enhancement of TNF-α during inflammatory conditions. Crush induces a large inflammatory reaction at the site of injury and increases TNF-α (George et al., 2004), and this increasing is reversed by thalidomide.
Background: It has been shown that chronic pain is able to induce depressive disorders in humans, in part, due to peripheral inflammation that reaches the central nervous system. However, the mechanisms involved remain to be established. The purpose of this study was to investigate whether sciatic nerve crush could produce depression-like behaviors, in addition to pain-related behaviors, in mice. Once confirmed, this model was used to investigate tumor necrosis factor-α (TNF-α) as a key mediator involved in the pathophysiology of both pain and depression. Experimental approach: Male Swiss mice were divided into three groups, naïve, sham and operated. In the operated group, the sciatic nerve was crushed. Following surgery, animals from the operated group were treated daily by oral gavage (p.o.) with saline (10 ml/kg), fluoxetine (20 mg/kg) or thalidomide (10 mg/kg) for 15 days. Mechanical hyperalgesia was evaluated every 3 days by von Frey filaments and depressive-like behavior was assessed at the end of day 15, using the tail suspension test (TST) and the forced swimming test (FST). Then, samples from the prefrontal cortex, hippocampus and sciatic nerve were processed to measure TNF-α levels by enzyme-linked immunosorbent assay (ELISA). Results: Crush caused significant mechanical hyperalgesia and depressive-like behaviors and increased TNF-α levels in the sciatic nerve, prefrontal cortex and hippocampus of operated animals. Treatment with fluoxetine or thalidomide reversed crush-induced mechanical hyperalgesia, depressive-like behaviors and the increased TNF-α levels in the sciatic nerve, prefrontal cortex and hippocampus. Conclusions: The sciatic nerve crush model represents a good model to study to mechanisms underlying both pain and depressive-like behaviors. Furthermore, inhibitors of TNF-α synthesis, like thalidomide, have a potential to treat depressive disorders associated with neuropathic pain.
Review of thalidomide use in the pediatric population
2015, Journal of the American Academy of Dermatology
Citation Excerpt :
Thalidomide has potent immunomodulatory, anti–tumor necrosis factor (TNF)-α, and antiangiogenic effects. It inhibits the chemotaxis and actions of immune cells, inhibits cytokine-induced nuclear factor kappa B transcription factor gene expression, and changes the interleukin milieu.1-4 It increases activity of suppressor T cells, decreases activity of helper T cells, and has variable effects on B-cell humoral immunity.5
Thalidomide is resurging in the management of adult rheumatologic skin conditions, especially lupus erythematosus. Although use in pediatric patients is reported since the 1990s, there are no systematic reviews describing treatment in children. Thalidomide has immunomodulatory and anti–tumor necrosis factor-α effects as well as antiangiogenic properties, making it useful for a broad spectrum of inflammatory disorders. Thalidomide is second-line treatment for aphthous stomatitis and chronic graft-versus-host disease in children and has been prescribed for many other conditions including actinic prurigo and epidermolysis bullosa pruriginosa. Systemic lupus erythematosus may be less responsive to thalidomide in children than adults. Peripheral neuropathy is observed in both idiosyncratic and dose-dependent relationships; children older than 12 years may be more susceptible to developing this adverse effect than younger patients. There are rare reports of thrombotic complications in children treated for nonmalignant indications. We review the mechanism of action and propose that thalidomide is an alternative treatment for patients who fail or have contraindications to anti–tumor necrosis factor-α biologics.

View all citing articles on Scopus

^☆: Thalidomide regulates TNF-α-induced NF-κB activation, but does not interrupt lipopolysaccharide-driven inflammatory cytokine production in monocytes isolated from patients with chronic granulomatous disease.

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Thalidomide attenuates excessive inflammation without interrupting lipopolysaccharide-driven inflammatory cytokine production in chronic granulomatous disease☆

Highlights

Abstract

Introduction

Section snippets

Monocyte isolation and stimulation

Thalidomide does not block caspase-1 activation or IL-1β production following LPS stimulation in CGD monocytes

Discussion

Conflict of interest

Acknowledgments

Blood

Burns

Cell. Immunol.

Blood

Gastroenterology

Cytokine

Immunol. Lett.

J. Biol. Chem.

Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica

Science

Osteomyelitis due to trimethoprim/sulfamethoxazole-resistant Edwardsiella tarda infection in a patient with X-linked chronic granulomatous disease

Infection

Inflammatory bowel disease in CGD reproduces the clinicopathological features of Crohn's disease

Am. J. Gastroenterol.

Complications of tumor necrosis factor-alpha blockade in chronic granulomatous disease-related colitis

Clin. Infect. Dis.