Figure - available from: Frontiers in Immunology
This content is subject to copyright.
Immunoglobulin G (IgG) isolated from amyotrophic lateral sclerosis (ALS) patients increase the production of nitric oxide (NO) and malondialdehyde (MDA) in BV-2 cells. NO production (A) and MDA production in BV-2 cells (B) were measured 24 h after treatments with control IgG and ALS IgG (0.1 mg/ml). NO production was measured indirectly using Griess assay and is expressed as NO2⁻ concentration. The results for each group are presented as mean ± SEM; ***p < 0.001 compared with untreated control group, ###p < 0.001 compared with the group treated with control IgG. The numbers in brackets indicate the number of different control IgG and ALS IgG samples examined and the number of untreated controls.
Source publication
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder with a very fast progression, no diagnostic tool for the presymptomatic phase, and still no effective treatment of the disease. Although ALS affects motor neurons, the overall pathophysiological condition points out to the non-cell autonomous mechanisms, where astrocytes and microg...
Similar publications
Diffuse Large B-cell lymphoma (DLBCL) is a highly aggressive disease with heterogeneous outcomes and marked variability in the response to chemotherapy. DLBCL comprises two major subtypes: germinal centre B-cell-like (GCB) and activated B-cell-like (ABC). Our study highlights the extensive antitumour activity of artesunate (ART) against both major...
Parkinson disease (PD) is the second-most common neurodegenerative disease. The characteristic pathology of progressive dopaminergic neuronal loss in people with PD is associated with iron accumulation and is suggested to be driven in part by the novel cell death pathway, ferroptosis. A unique modality of cell death, ferroptosis is mediated by iron...
Citations
... Microglial cells in culture are affected by ALS IgG in a less apparent way. Namely, a BV-2 microglial cell line displayed a rise in the signal from fluorescent markers of oxidative stress in response to the application of only 4/11 ALS IgG patient samples 15 . It is well known that microglia participate in many neuroinflammatory pathologies, adding to oxidative stress and late progression phase in the non-cell autonomous mechanism of ALS 16 , 17 . ...
... An example was shown here that microglial cells did not respond to ALS IgG with the same vigor as astrocytes. This is in line with the finding of the scarce generation of peroxide in the microglial cell line upon ALS IgG treatment 15 . ...
... Metabolic diseases are a new field of study, and the evidence currently available is insufficient to draw any conclusions about the possible harm to humans. However, the present evidence suggests that more research into the effects of BPA on adiposity, glucose or insulin control, lipids, and other diabetes or metabolic syndrome end-points is needed [19]. ...
Bisphenols (BP) is one of the most important and highest volumes of chemicals produced in the universe. Each year, around 100 tons of bisphenol compounds are released into the atmosphere. In general, bisphenol is most widely used for production of polycarbonate (making plastic bottles like baby bottles and nursing products, dental sealants, CDs, DVDs, eye glasses, medical equipment’s, plasticizers etc.) and polymeric resins (epoxy resins, impact resistant safety materials like sports goods etc.). Due to these unavoidable chemicals, human beings are affected by human chronic diseases like obesity, toxicity, neuro disorder, reproductivity disorders, diabetes, cardio related issues, birth defects, metabolic syndrome, breathing issues, digestive related issues, cancer, genetic mutation etc., Children are easily affecting due to the multi dose consumption of packed food containing BP (canned foods) than adults. Women are affecting polycystic ovaries due to the high-level deposition of BP.
... Proposedly, misfolded SOD1 proteins are deposited on organelles, especially mitochondria, and impair mitochondrial function [7][8][9], and then, the dysfunctional mitochondria cause cellular ROS accumulation, serving as a key mediator in a vicious cycle of oxidative stress and exacerbated mitochondrial dysfunction [10]. Accumulated reactive oxygen species (ROS), such as hydrogen peroxide and hydroxyl free radicals, play a critical role in mediating oxidative stress in ALS, resulting in lipid peroxidation and genotoxicity [11]. ...
Background:
Oxidative stress and reactive oxygen species (ROS) are important in the pathogenesis of amyotrophic lateral sclerosis (ALS). Hypochlorous acid (HOCl) is a powerful oxidant of the reactive oxygen species (ROS) family. HOCl's role in the progress of ALS remains unclear due to the lack of an effective HOCl detection method. Cumulative evidence supports oxidative damage incurred by mutant hSOD1 contributing to motor neuron death; however, whether HOCl as well as its catalytic enzyme myeloperoxidase (MPO) function in the cell death of SOD1G93A ALS remains elusive.
Methods:
The hSOD1WT and hSOD1G93A NSC-34 cell and SOD1G93A ALS mouse models were employed. With a novel fluorescent HOCl probe, HKOCl-3, we detected the expressions of HOCl and its catalytic enzyme, MPO, in the above models in vitro and in vivo. The regulation of MPO/HOCl by hSOD1G93A mutation and cell deaths by MPO/HOCl were also assayed, including apoptosis, ferroptosis, and autophagy.
Results:
Our results showed that hSOD1G93A mutation promoted the activation of the MPO/HOCl pathway in SOD1G93A ALS cell models. The activation of MPO/HOCl pathways facilitated apoptosis and ferroptosis through increasing the Bax/Bcl-2 ratio and expression of caspase-3 or inhibiting the expressions of GPX4 and NQO1 and thus leading to irreversible lipid peroxidation. Overexpressed FSP1, a glutathione-independent suppressor, could ameliorate ferroptosis. In vivo, we demonstrated that the activation of the MPO/HOCl pathway occurred differently in motor neurons of the motor cortices, brain stems, and spinal cords in male and female SOD1G93A transgenic mice. In addition, inhibiting MPO improved the motor performance of SOD1G93A transgenic mice, as demonstrated by the rotarod test.
Conclusions:
We concluded that aggregation of mutant hSOD1 proteins contributed to activation of the MPO/HOCl pathway, triggering apoptosis and ferroptosis in motor neuronal deaths and exerting impaired motor performance.
... On the one hand, it comes from the presence of numerous autoimmune IgG antibodies, characterized by the examination of sera from ALS patients [41]. Besides the autoimmune properties, these antibodies can elevate the motoneuronal calcium level [42,43] and induce selective MN apoptosis in rat mixed primary spinal cord cultures [44]. Immunoglobulins from patients with sporadic ALS can bind and modify the function of L-type, P-type, and other neuronal calcium channels [45][46][47], which increase the frequency of miniature end-plate potentials eventually [30]. ...
Introduction: Previously, we demonstrated the degeneration of axon terminals in mice after repeated injections of blood sera from amyotrophic lateral sclerosis (ALS) patients with identified mutations. However, whether a similar treatment affects the cell body of motor neurons (MNs) remained unresolved. Methods: Sera from healthy individuals or ALS patients with a mutation in different ALS-related genes were intraperitoneally injected into ten-week-old male Balb/c mice (n = 3/serum) for two days. Afterward, the perikaryal calcium level was measured using electron microscopy. Furthermore, the optical disector method was used to evaluate the number of lumbar MNs. Results: The cytoplasmic calcium level of the lumbar MNs of the ALS-serum-treated mice, compared to untreated and healthy-serum-treated controls, was significantly elevated. While injections of the healthy serum did not reduce the number of MNs compared to the untreated control group, ALS sera induced a remarkable loss of MNs. Discussion: Similarly to the distant motor axon terminals, the injection of blood sera of ALS patients has a rapid degenerative effect on MNs. Analogously, the magnitude of the evoked changes was specific to the type of mutation; furthermore, the degeneration was most pronounced in the group treated with sera from ALS patients with a mutation in the chromosome 9 open reading frame 72 gene.
... Intriguingly, immunoglobulin G (IgG) (caption on next page) F. De Marchi et al. has been reported to accumulate in and/or around motor neurons (Donnenfeld et al., 1984;Engelhardt and Appel, 1990). Moreover, administration of IgG from ALS patients can be taken up and/or stimulate motor neurons, astrocytes, and microglia, increase serum and spinal cord cytokine levels, and elicit motor neuron degeneration (Appel et al., 1991;Bataveljic et al., 2014;Milošević et al., 2017;Obál et al., 2016;Pagani et al., 2011;Pullen et al., 2004). Some of the antibodies recognize aggregated proteins and neurofilaments, but their exact pathogenic mechanism is unclear (Malaspina et al., 2015;May et al., 2014). ...
Amyotrophic lateral sclerosis (ALS) is a debilitating and rapidly fatal neurodegenerative disease. Despite decades of research and many new insights into disease biology over the 150 years since the disease was first described, causative pathogenic mechanisms in ALS remain poorly understood, especially in sporadic cases. Our understanding of the role of the immune system in ALS pathophysiology, however, is rapidly expanding. The aim of this manuscript is to summarize the recent advances regarding the immune system involvement in ALS, with particular attention to clinical translation. We focus on the potential pathophysiologic mechanism of the immune system in ALS, discussing local and systemic factors (blood, cerebrospinal fluid, and microbiota) that influence ALS onset and progression in animal models and people. We also explore the potential of Positron Emission Tomography to detect neuroinflammation in vivo, and discuss ongoing clinical trials of therapies targeting the immune system. With validation in human patients, new evidence in this emerging field will serve to identify novel therapeutic targets and provide realistic hope for personalized treatment strategies.
... Out of these, IgG, also present in post-mortem ALS samples, has been found to favour neurodegeneration, both in vitro and in vivo [49,155,156]. IgG further possesses pro-inflammatory properties, promoting microglia recruitment and upregulation of inflammatory cytokines [124,141,142]. Some of the other chemokines found to be elevated in ALS-CSF include IL-6, TNF-α and TGF-β [81,130,172], with the finding of raised TNF-α levels being particularly interesting, given its associations with glutamate excitotoxicity [43,178,218]. ...
Abstract Despite the considerable progress made towards understanding ALS pathophysiology, several key features of ALS remain unexplained, from its aetiology to its epidemiological aspects. The glymphatic system, which has recently been recognised as a major clearance pathway for the brain, has received considerable attention in several neurological conditions, particularly Alzheimer’s disease. Its significance in ALS has, however, been little addressed. This perspective article therefore aims to assess the possibility of CSF contribution in ALS by considering various lines of evidence, including the abnormal composition of ALS-CSF, its toxicity and the evidence for impaired CSF dynamics in ALS patients. We also describe a potential role for CSF circulation in determining disease spread as well as the importance of CSF dynamics in ALS neurotherapeutics. We propose that a CSF model could potentially offer additional avenues to explore currently unexplained features of ALS, ultimately leading to new treatment options for people with ALS.
... Besides the effect on motor neurons, the various specific effects of ALS IgG were described in other cell types, also playing a role in the pathomechanism of ALS. These antibodies are capable of inducing oxidative stress and an upregulation of the antioxidative system in a BV-2 microglial cell line [53] and also affect cytosolic calcium homeostasis in cultured rat astrocytes [54]. Nevertheless, sera from two sporadic ALS patients were also included in the present study, which proved that identical results could be obtained from these patients as in the previous study. ...
Previously, we demonstrated increased calcium levels and synaptic vesicle densities in the motor axon terminals (MATs) of sporadic amyotrophic lateral sclerosis (ALS) patients. Such alterations could be conferred to mice with an intraperitoneal injection of sera from these patients or with purified immunoglobulin G. Later, we confirmed the presence of similar alterations in the superoxide dismutase 1 G93A transgenic mouse strain model of familial ALS. These consistent observations suggested that calcium plays a central role in the pathomechanism of ALS. This may be further reinforced by completing a similar analytical study of the MATs of ALS patients with identified mutations. However, due to the low yield of muscle biopsy samples containing MATs, and the low incidence of ALS patients with the identified mutations, these examinations are not technically feasible. Alternatively, a passive transfer of sera from ALS patients with known mutations was used, and the MATs of the inoculated mice were tested for alterations in their calcium homeostasis and synaptic activity. Patients with 11 different ALS-related mutations participated in the study. Intraperitoneal injection of sera from these patients on two consecutive days resulted in elevated intracellular calcium levels and increased vesicle densities in the MATs of mice, which is comparable to the effect of the passive transfer from sporadic patients. Our results support the idea that the pathomechanism underlying the identical manifestation of the disease with or without identified mutations is based on a common final pathway, in which increasing calcium levels play a central role.
... Notably, patients with pediatric OMS exhibit increased expression of a microglial marker and proinflammatory cytokines in cerebrospinal fluid (CSF) and some children with OMS are post-infectious [9][10][11]. Furthermore, serum IgG or autoantibody existed in patients enhances microglial activation in Parkinson disease (PD) [12], amyotrophic lateral sclerosis (ALS) [13], and systemic lupus erythematosus (SLE) [14][15][16]. Thus, it is reasonable to hypothesize that serum IgG from children with OMS and NB may impact the activation of microglia. ...
... Moreover, the expression of microglial marker soluble CD14 and proinflammatory cytokines is enhanced in CSF from patients with pediatric OMS [10,11]. Additionally, serum IgG from patients or autoantibody existed in patients directly enhances microglial activation in PD [12], ALS [13], and SLE [14][15][16] or initially binds with astrocytes or neurons and further indirectly affects microglial activation [42,43]. Autoantibodies are also detectable in sera and CSF of children with OMS [25,26] and may be contained in serum IgG from children with OMS and NB in our study, although we did not identify these autoantibodies and autoantibodies include IgM besides IgG. ...
... Consistently, we and others found that IgG-induced neuronal cytolysis occurs in pediatric OMS rather than adult OMS [6,7]. Notably, increased microglial activation is not specific to only OMS, since previous literatures have documented that serum IgG from patients with PD [12] or ALS [13] enhances the activation of microglia and the production of NO, and serum IgG from patients with SLE induced behavioral changes is mediated by microglial activation [14][15][16]. Moreover, several autoantibodies found in patients with OMS [44][45][46][47] also exist in other diseases, such as autoantibody against glycine receptor in progressive encephalomyelitis with rigidity and myoclonus [46,48] or autoantibody against glutamic acid decarboxylase in stiff-person syndrome [48]. ...
Background:
Opsoclonus-myoclonus syndrome (OMS) is a rare neurological disease. Some children with OMS also have neuroblastoma (NB). We and others have previously documented that serum IgG from children with OMS and NB induces neuronal cytolysis and activates several signaling pathways. However, the mechanisms underlying OMS remain unclear. Here, we investigated whether nitric oxide (NO) from activated microglias and its cascade contribute to neuronal cytolysis in pediatric OMS.
Methods:
The activation of cultured cerebral cortical and cerebellar microglias incubated with sera or IgG isolated from sera of children with OMS and NB was measured by the expression of the activation marker, cytokines, and NO. Neuronal cytolysis was determined after exposing to IgG-treated microglia-conditioned media. Using inhibitors and activators, the effects of NO synthesis and its intracellular cascade, namely soluble guanylyl cyclase (sGC) and protein kinase G (PKG), on neuronal cytolysis were evaluated.
Results:
Incubation with sera or IgG from children with OMS and NB increased the activation of cerebral cortical and cerebellar microglias, but not the activation of astrocytes or the cytolysis of glial cells. Moreover, the cytolysis of neurons was elevated by conditioned media from microglias incubated with IgG from children with OMS and NB. Furthermore, the expression of NO, sGC, and PKG was increased. Neuronal cytolysis was relieved by the inhibitors of NO signaling, while neuronal cytolysis was exacerbated by the activators of NO signaling but not proinflammatory cytokines. The cytolysis of neurons was suppressed by pretreatment with the microglial inhibitor minocycline, a clinically tested drug. Finally, increased microglial activation did not depend on the Fab fragment of serum IgG.
Conclusions:
Serum IgG from children with OMS and NB potentiates microglial activation, which induces neuronal cytolysis through the NO/sGC/PKG pathway, suggesting an applicability of microglial inhibitor as a therapeutic candidate.
... Macrophages are known to play a critical role in innate immunity, and attack, destroy and ingest foreign substances through the production of cytokines and NO (47), primarily because NO can activate the immune system, and can react with the superoxide anions produced by the respiratory burst of phagocytes to generate a strong oxidant-peroxynitrite (ONOOH), which can kill or inhibit the growth of several pathogenic microorganisms. Therefore, NO serves as a critical cellular defense factor in the anti-infective immunity of an organism (48,49), and NO synthesis is an important mechanism underlying the non-specific immunity of macrophages (50). For teleosts, which rely on non-specific immune processes to enhance their immune response and resist pathogen infection (51), cytokines such as TNF-α, IL-1β, IL-6, IL-10, IL-12, and TGF-β, primarily produced by activated macrophages, also play an important role in the immune system. ...
The effects of the oral administration of Rehmannia glutinosa polysaccharide (RGP-1) on the immunoregulatory properties, antioxidant activity, and resistance against Aeromonas hydrophila in Cyprinus carpio L. were investigated. The purified RGP-1 (250, 500, and 1,000 μg/mL) was co-cultured with the head kidney cells of the common carp. The proliferation and phagocytosis activities of the head kidney cells, and the concentration of nitric oxide (NO) and cytokines in the culture medium were determined. Next, 300 common carps (47.66 ± 0.43 g) were randomly divided into five groups; the two control groups (negative and positive) were administered sterile PBS and the three treatment groups were administered different concentrations of RGP-1 (250, 500, and 1,000 μg/mL) for seven days. Subsequently, the positive and treatment groups were infected with A. hydrophila, and the negative group was administered sterile PBS for 24 h. The concentration of NO, cytokines, lysozyme (LZM), and alkaline phosphatase (AKP) in serum, the total antioxidant capacity (T-AOC), the levels of malonaldehyde (MDA) and glutathione (GSH), and the total activities of superoxide dismutase (T-SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in the hepatopancreas of the common carp were tested. We observed that RGP-1 could significantly enhance the proliferation and phagocytosis activities (P < 0.05), besides inducing the production of NO, pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-12) and anti-inflammatory cytokines (IL-10, TGF-β) (P < 0.05) in vitro. The in vivo experimental results revealed that RGP-1 significantly enhanced NO production, protein levels of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-12), LZM and AKP activities, and the antioxidant content (T-AOC, SOD, CAT, GSH, GSH-Px, and MDA) compared to that observed in the negative group prior to A. hydrophila infection (P < 0.05). NO, pro-inflammatory cytokines, LZM and AKP activities were significantly lower than that in the positive group after infection (P < 0.05). However, whether infected or not, the expression of anti-inflammatory cytokines (IL-10, TGF-β) increased significantly in the RGP-1-treated groups (P < 0.05). Therefore, the results suggested that RGP-1 could enhance the non-specific immunity, antioxidant activity and anti-A. hydrophila activity of the common carp, and could be used as a safe and effective feed additive in aquaculture.
... The effect of purified IgG on motoneuronal calcium accumulation was demonstrated in other laboratories, as well [22,23], furthermore various specific effects of ALS IgG were described related to other cell types playing a role in the pathomechanism of ALS. ALS IgG was shown to induce oxidative stress and upregulation of the antioxidative system in a BV-2 microglial cell line [24]. It was also shown to affect cytosolic Ca-homeostasis in cultured rat astrocytes [25]. ...
In an earlier study, signs of commencing degeneration of spinal motor neurons were induced in mice with short-term intraperitoneal injections of immunoglobulin G (IgG) taken from patients with amyotrophic lateral sclerosis (ALS). Since in that study, neither weakness nor loss of motor neurons was noted, to test whether the ALS IgG in this paradigm has the potential to evoke relentless degeneration of motor neurons, treatment with repeated injections over a longer period was carried out. Mice were systematically injected intraperitoneally with serum taken from ALS patients over a 75-day period. At selected time points, the isometric force of the limbs, number of spinal motor neurons and their intracellular calcium levels were determined. Furthermore, markers of glial activation and the motoneuronal uptake of human IgG were monitored. During this period, gliosis and progressive motoneuronal degeneration developed, which led to gradual loss of spinal motor neurons, more than 40% at day 21, along with decreasing muscle strength in the limbs. The inclusion-like accumulation of IgG appeared in the perikarya with the increase of intracellular calcium in the cell bodies and motor nerve terminals. Our results demonstrate that ALS serum can transfer motor neuron disease to mice.
