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Neuromyelitis optica, a rare neuroinflammatory demyelinating disease of the CNS, is characterized by the presence of specific pathogenic autoantibodies directed against the astrocytic water channel aquaporin 4 (AQP4) and is now considered as an astrocytopathy associated either with complement-dependent astrocyte death or with astrocyte dysfunction. However, the link between astrocyte dysfunction and demyelination remains unclear. We propose glial intercellular communication, supported by connexin hemichannels and gap junctions, to be involved in demyelination process in neuromyelitis optica. Using mature myelinated cultures, we demonstrate that a treatment of 1 h to 48 h with immunoglobulins purified from patients with neuromyelitis optica (NMO-IgG) is responsible for a complement independent demyelination, compared to healthy donors’ immunoglobulins (P

Background: Blood-brain barrier (BBB) breakdown is considered one of the key steps for the development and lesion formation of neuromyelitis optica spectrum disorder (NMOSD). However, little is known about the molecular mechanisms involved. The aim of the study was to evaluate the effect of human immunoglobulins from NMOSD patients (NMO-IgG) on BBB properties.Methods: Freshly isolated brain microvessels (IBM) from rat brains that mirror the blood-CNS barrier were used as a study model. At first, analysis of the secretome profile (cytokine protein array) from IBM exposed to purified NMO-IgG, to healthy donor IgG (Control-IgG), or non-treated, was performed. Second, structural BBB properties were analysed by Western blotting (Wb) to measure the expression of tight junction (TJ) proteins (Claudin-5, occludin, and ZO-1) in fresh IBM and primary cultures of brain microvascular endothelial cells (BMEC) after exposition to NMO-IgG and Control-IgG. Finally, functional BBB properties were investigated using two approaches; 1) in the NMO-rat model, the presence of rat-IgG in tissue lysate from brain periventricular regions was analysed using Wb, and 2) in a bicameral model simulating the blood and brain compartments of the BBB, the passage of NMO-IgG and sucrose was assessed by using ELISA.Results: We found that NMO-IgG induces several functional and morphological changes of the BBB in our different study models, including: 1) increase of pro-inflammatory cytokines production (CXCL-10 [IP-10], IL-6, IL1-ra, IL1-β and CXCL-3) by at least three-fold in IBM when exposed to NMO-IgG in comparison to Control-IgG or non-treated IBM; 2) decrease of Claudin-5 levels by 25.6% after treatment of fresh IBM by NMO-IgG compared to Control-IgG (p=0.002), and similarly, decrease of Claudin-5 by at least 20% when BMEC were cultured with NMO-IgG from five different patients; 3) a higher level of rat-IgG accumulated in periventricular regions of NMO-rats compared to Control-rats and an increase the permeability of BBB after NMO-IgG treatment in the bicameral model.Conclusion: Human NMO-IgG induces both structural and functional alterations of BBB properties, suggesting a direct role of NMO-IgG on modulation of BBB permeability in NMOSD.

Our knowledge of multiple sclerosis (MS) neuropathology has benefited from a number of studies that provided an in-depth description of plaques and, more recently, diffuse alterations of the normal-appearing white or grey matter. However, there have been few studies focusing on the periplaque regions surrounding demyelinated plaques, notably in MS spinal cords. In this context, the present study aimed to analyze the molecular immunopathology of periplaque demyelinated lesions (PDLs) in the spinal cord of patients with a progressive form of MS. To achieve this goal, the neuropathological features of PDLs were analyzed in postmortem tissues derived from the cervical spinal cord of 21 patients with primary or secondary progressive MS. We found that PDLs covered unexpectedly large areas of incomplete demyelination and were characterized by the superimposition of pro- and anti-inflammatory molecular signatures. Accordingly, macrophages/microglia accumulated in PDLs but exhibited a poor phagocytic activity toward myelin debris. Interestingly, while genes of the oligodendrocyte lineage were consistently down-regulated in PDLs, astrocyte-related molecules such as aquaporin 4, connexin 43 and the glutamate transporter EAAT1, were significantly upregulated in PDLs at the mRNA and protein levels. Overall, our work indicates that in the spinal cord of patients with a progressive form of MS, a tissue remodeling process that is temporally remote from plaque development takes place in PDLs. We propose that in spinal cord PDLs, this process is supported by subtle alterations of astrocyte functions and by low-grade inflammatory events that drive a slowly progressive loss of myelin and a failure of remyelination.

Background Devic’s neuromyelitis optica (NMO) is an autoimmune astrocytopathy, associated with central nervous system inflammation, demyelination, and neuronal injury. Several studies confirmed that autoantibodies directed against aquaporin-4 (AQP4-IgG) are relevant in the pathogenesis of NMO, mainly through complement-dependent toxicity leading to astrocyte death. However, the effect of the autoantibody per se and the exact role of intrathecal AQP4-IgG are still controversial. Methods To explore the intrinsic effect of intrathecal AQP4-IgG, independent from additional inflammatory effector mechanisms, and to evaluate its clinical impact, we developed a new animal model, based on a prolonged infusion of purified immunoglobulins from NMO patient (IgGAQP4+, NMO-rat) and healthy individual as control (Control-rat) in the cerebrospinal fluid (CSF) of live rats. Results We showed that CSF infusion of purified immunoglobulins led to diffusion in the brain, spinal cord, and optic nerves, the targeted structures in NMO. This was associated with astrocyte alteration in NMO-rats characterized by loss of aquaporin-4 expression in the spinal cord and the optic nerves compared to the Control-rats (p = 0.001 and p = 0.02, respectively). In addition, glutamate uptake tested on vigil rats was dramatically reduced in NMO-rats (p = 0.001) suggesting that astrocytopathy occurred in response to AQP4-IgG diffusion. In parallel, myelin was altered, as shown by the decrease of myelin basic protein staining by up to 46 and 22 % in the gray and white matter of the NMO-rats spinal cord, respectively (p = 0.03). Loss of neurofilament positive axons in NMO-rats (p = 0.003) revealed alteration of axonal integrity. Then, we investigated the clinical consequences of such alterations on the motor behavior of the NMO-rats. In a rotarod test, NMO-rats performance was lower compared to the controls (p = 0.0182). AQP4 expression, and myelin and axonal integrity were preserved in AQP4-IgG-depleted condition. We did not find a major immune cell infiltration and microglial activation nor complement deposition in the central nervous system, in our model. Conclusions We establish a link between motor-deficit, NMO-like lesions and astrocytopathy mediated by intrathecal AQP4-IgG. Our study validates the concept of the intrinsic effect of autoantibody against surface antigens and offers a model for testing antibody and astrocyte-targeted therapies in NMO. Electronic supplementary material The online version of this article (doi:10.1186/s12974-016-0577-8) contains supplementary material, which is available to authorized users.

Obesity results from disturbances of tightly regulated interactions between the nervous, endocrine, and metabolic systems that can be caused by external factors, such as viral infections. A mouse model of obesity induced by brain infection with a morbillivirus, canine distemper virus, allowed us to identify obesity-related genes. Using a subtractive library for the hypothalamus, the main brain structure regulating energy homeostasis, we identified a new gene on mouse chromosome 19 which we named upregulated obese product (Urop) 11 and, which has no homology with any known mRNA. A step-by-step molecular approach allowed us to isolate the full-length mRNA, predict the protein sequence, and identify consensus sites. Urop11 was mainly detected in the hypothalamus and adipocytes, and was dramatically upregulated in these central and peripheral structures in obese mice. Urop11 was also expressed in human neural and lymphoid samples and its expression seemed to be regulated by the state of lymphocyte activation. Interestingly, Urop11 expression was strongly upregulated both in vivo in mouse hypothalamus and in vitro in mouse neural cell lines, after leptin treatment. Taken together, our data show that Urop11 is a target of leptin, the satiety factor produced by adipocytes, in physiological and pathological conditions, including obesity. This new gene can be considered a key molecule in the hypothalamic integration pathway and demonstrates the importance of Urop11 as a target of leptin action.

The semaphorin-signaling transducer collapsin response mediator protein 2 (CRMP2) has been identified in the nervous system where it mediates Sema3A-induced growth cone navigation. In the present study, we provide first evidence that CRMP2 is present in the immune system and plays a critical role in T lymphocyte function. CRMP2 redistribution at the uropod in polarized T cells, a structural support of lymphocyte motility, suggests that it may regulate T cell migration. This was evidenced in primary T cells by small-interfering RNA-mediated CRMP2 gene silencing and blocking Ab, as well as CRMP2 overexpression in Jurkat T cells tested in a chemokine- and semaphorin-mediated transmigration assay. Expression analysis in PBMC from healthy donors showed that CRMP2 is enhanced in cell subsets bearing the activation markers CD69+ and HLA-DR+. Heightened expression in T lymphocytes of patients suffering from neuroinflammatory disease with enhanced T cell-transmigrating activity points to a role for CRMP2 in pathogenesis. The elucidation of the signals and mechanisms that control this pathway will lead to a better understanding of T cell trafficking in physiological and pathological situations.

The aim of this study was to investigate interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) production by peripheral blood mononuclear cells (PBMC) in response to oral challenge in patients with food allergy. Thirty-one patients were compared with 10 healthy subjects. Cell cultures were prepared before and 150 min after single-blind, placebo-controlled, oral food challenge. In vitro production of cytokines was observed after stimulation with phytohemagglutinin (PHA) plus phorbol myristate acetate (PMA). Patients were characterized according to their reactions. In vitro IL-4 production was significantly greater in patients with food sensitization than in controls. The 18 patients with positive food challenge presented significantly greater IL-4 production after food challenge than before. IFN-gamma production was not modified. In the 10 control subjects and in 13 patients with negative food challenge, IL-4 and IFN-gamma production was comparable before and after food challenge. The increased IL-4 production in patients with positive oral food challenge could account for the development of polysensitization. This situation might be relevant to other allergic diseases and to treatments.

The fragment of the Fc segment of IgE resistant to proteolytic enzymes was determined by a radioimmunologic method in fecal extracts from several groups of patients. IgEs remained undetectable in the 95 healthy subjects studied. IgEs were detected in 16/27 carriers of intestinal parasites (60%), with a mean of 92.4 IU/g dry weight. IgEs were also detected in 236/312 food-sensitization patients, with the sensitizing foodstuffs being identified by searching for the specific IgEs in circulation (75%), with a mean of 63.9 IU/g dry weight. The simultaneous determination of alpha-1-antitrypsin in fecal extracts and in sera from 21 control subjects and 21 food-sensitization carriers demonstrated that extravasation of plasma proteins into the gut lumen cannot be responsible for the presence of IgE in stools. Testing for IgE in stools therefore appears to be a simple and economical means of identifying patients without parasites who present with food sensitization.

Lymphocyte migration into the central nervous system is a critical step in the physiopathology of a variety of neurological diseases, including multiple sclerosis and virus-induced neuroinflammation. To better understand the molecular mechanisms involved in cells migration, we focused our studies on collapsin response mediator proteins (CRMPs), a group of phosphoproteins that mediate neural cell motility. There is now evidence that collapsin response mediator protein 2 (CRMP2) plays critical roles in the polarization (uropod formation) of T lymphocytes and their subsequent migration. CRMP2 was known to respond to semaphorin, ephrin and neurotrophin signaling in neurons. The link between the chemokine CXCL12, CRMP2 activity and cell migration has been demonstrated in T lymphocytes. These observations and comparisons of the activity of CRMPs in immune and non-immmune cells are summarized here. The ability of a human retrovirus to enhance lymphocyte migration through the modulation of CRMP2 activity is also discussed. In conclusion, viruses have the ability to manipulate the lymphocyte motility machinery, intensifying neural tissue invasion in infected patients.

Food allergy is a group of distinct clinico-pathological entities that have an immunological basis in common, and in which an abnormal or exaggerated immunological response to a specific food leads to disease. Some clinical pictures involving multiple organ system (anaphylaxis) are potentially fatal. The data on the incidence, prevalence, mortality rate and food products involved in food-induced anaphylaxis and the evolution of food sensitization compared with changes in eating habits are not very reliable. In the present study we analysed, over a period of 9 years (1984-1992), a group of 580 patients with pathological reactions to foods, 60 of which presented severe, near-fatal reactions. We sought the etiologic components and food sensitization in comparison with the principal tendencies of food consumption in France. Food products most frequently incriminated in anaphylactic reactions are not of a primary nutritional importance: celery (30%), crustaceans (17%), fish (13%), peanuts (12%), mango (6%), mustard (3%), but they are often hidden allergens in commercial foods. The sensitization to food products in the group of 580 patients reveals, in decreasing order of frequency: wheat (39%), peanuts (37%), crab (34%), celery (30%), soy (30%). Compared with previous data, the frequency of sensitization to different foods has changed; for instance, the sensitizations to wheat, soy, peanuts, celery, mustard, rice, are definitely increasing. The increased consumption and more attentive clinical research may be the reasons for this evolution. For products such as egg and pork, the data are stable and parallel with consumption, whilst for other products like milk and other dairy products, the increased consumption is accompanied by a decrease of the incidence of sensitization. The reactivity to some allergens may be affected by the way of preparing the food.

Resume Le denombrement des cellules contenant des immunoglobulines a ete effectue par immunofluorescence au sein des muqueuses gastrique, duodenale et jejunale chez 120 sujets sains et 200 malades chez lesquels une sensibilisation alimentaire avait ete mise en evidence par la detection des IgE specifiques plasmatiques et la reactivite intestinale par une epreuve de provocation orale. Aucun sujet sain et aucun malade n'etait porteur de parasite. Les nombres de cellules contenant de l'IgA, de l'IgM, de l'IgG ou de l'IgD etaient semblables chez les malades et chez les temoins. Chez les allergiques les cellules renfermant de l'IgE presentes dans la lamina propria de l'intestin etaient systematiquement en nombre tres accru. Ceci a ete observe en ce qui concerne la muqueuse gastrique chez seulement 118 des 200 malades. Cinquante autres malades suspectes de presenter une allergie alimentaire et presentant un accroissement de la densite des cellules duodenales contenant de l'IgE ont ete traites par le cromoglycate disodique oral pendant un minimum de trois mois. Trente-trois malades ont observe une guerison ou une amelioration marquee de leurs symptomes. Le denombrement des cellules duodenales contenant de l'IgE, mais non celui des cellules de l'estomac, represente un moyen efficace de diagnostiquer une allergie alimentaire chez des sujets exempts de parasites intestinaux.

Devic's neuromyelitis optica is an inflammatory demyelinating disorder normally restricted to the optic nerves and spinal cord. Since the identification of a specific autoantibody directed against aquaporin 4, neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody, neuromyelitis optica has been considered an entity distinct from multiple sclerosis. Recent findings indicate that the neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody has a pathogenic role through complement-dependent astrocyte toxicity. However, the link with demyelination remains elusive. Autoantibodies can act as receptor agonists/antagonists or alter antigen density in their target cells. We hypothesized that the neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody impairs astrocytic function and secondarily leads to demyelination. Rat astrocytes and oligodendrocytes from primary cultures and rat optic nerves were exposed long-term (24 h) to immunoglobulin G in the absence of complement. Immunoglobulin G was purified from the serum of patients with neuromyelitis optica who were either neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody positive or negative, as well as from healthy controls. Flow cytometry analysis showed a reduction of membrane aquaporin 4 and glutamate transporter type 1 on astrocytes following contact with immunoglobulin G purified from neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody positive serum only. The activity of glutamine synthetase, an astrocyte enzyme converting glutamate into glutamine, decreased in parallel, indicating astrocyte dysfunction. Treatment also reduced oligodendrocytic cell processes and approximately 30% oligodendrocytes died. This deleterious effect was confirmed ex vivo; exposed optic nerves showed reduction of myelin basic protein. Immunoglobulin G from neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody seronegative patients and from healthy controls had no similar effect. Neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody did not directly injure oligodendrocytes cultured without astrocytes. A toxic bystander effect of astrocytes damaged by neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody on oligodendrocytes was identified. Progressive accumulation of glutamate in the culture medium of neuromyelitis optica-immunoglobulin G/aquaporin 4-antibody-treated glial cells supported the hypothesis of a glutamate-mediated excitotoxic death of oligodendrocytes in our models. Moreover, co-treatment of glial cultures with neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody and d+2-amino-5-phosphonopentanoic acid, a competitive antagonist at the N-methyl-d-aspartate/glutamate receptor, partially protected oligodendrocytes. Co-immunolabelling of oligodendrocyte markers and neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody showed that astrocytic positive processes were in close contact with oligodendrocytes and myelin in rat optic nerves and spinal cord, but far less so in other parts of the central nervous system. This suggests a bystander effect of neuromyelitis optica-immunoglobulin G-damaged astrocytes on oligodendrocytes in the nervous tissues affected by neuromyelitis optica. In conclusion, in these cell culture models we found a direct, complement-independent effect of neuromyelitis optica-immunoglobulin G/aquaporin 4 antibody on astrocytes, with secondary damage to oligodendrocytes possibly resulting from glutamate-mediated excitotoxicity. These mechanisms could add to the complement-induced damage, particularly the demyelination, seen in vivo.

Collapsin Response Mediator Protein (CRMP)-2 is involved in T-cell polarization and migration. To address the role of CRMP2 in neuroinflammation, we analyzed its involvement in lymphocyte recruitment to the central nervous system in mouse infected with neurotropic and non-neurotropic virus strains (RABV, CDV). A sub-population of early-activated CD69+CD3+ T lymphocytes highly expressing CRMP2 (CRMP2hi) peaked in the blood, lymph nodes and brain of mice infected with neurotropic viruses, and correlated with severity of disease. They displayed high migratory properties reduced by CRMP2 blocking antibody. These data point out the potential use of CRMP2 as a peripheral indicator of neuroinflammation.

Background: Although fish gelatin may represent a useful alternative to bovine or porcine gelatins, the clearly recognized high prevalence of fish allergy could increase the risk of anaphylaxis to gelatin. The rationale for investigating tuna gelatin rather than gelatins from more allergenic fishes is the availability of an industrial gelatin under development. The infrequent occurrence of tuna allergy should influence the safety of a derived product. The present study investigated IgE antibodies to tuna-skin-derived gelatin in adults and children with documented fish allergy or sensitization. Methods: Serum samples were taken from 100 consecutive patients with fish allergy or sensitization and tested for IgE antibodies against hydrolyzed or nonhydrolyzed gelatin extracted from tuna skin as compared to extracts from tuna flesh, tuna skin as well as bovine or porcine gelatins. Patients with tuna allergies or sensitization were sensitive to the same tuna species (yellowfin) as that from which the gelatin was obtained. IgE antibodies to these various extracts were analyzed using SDS-PAGE and immunoblotting. Results: Only 3 of the 100 serum samples tested gave evidence of reactivity to gelatin extracted from tuna skin. Cross-reactivity between bovine/porcine and fish gelatins was not observed. Conclusion: The risk of adverse reactions to tuna skin gelatin seems to be significantly lower than the risk of fish allergy. Fish gelatin may represent a valuable alternative to bovine or porcine gelatins.

Intestinal absorption of mannitol (used as a marker of normal monomer passage) and intestinal absorption of lactulose (used as a marker of macromolecule exclusion) were measured in 15 healthy subjects and in 20 patients with food allergy. When studied in the fasting state, there was no apparent difference in the absorption of mannitol between control subjects and allergic patients although the absorption of lactulose was more marked, but not significantly, in the latter group. Ingestion of an allergen by these patients led to a significant increase in the absorption of lactulose (intestinal passage increased by a factor of two). Administration of 300 mg cromoglycate 1/4 hour before the provocation test completely averted abnormal intestinal permeability. Digestive absorption of cromoglycate was also studied in the same 15 control subjects and 20 allergic patients. The peak plasma concentration and area under the curve were significantly greater in healthy subjects when the study was carried out in the fasting state. The application of an oral provocation test with an allergen also significantly increased the plasma concentration of cromoglycate in allergic patients. Contact between an allergen and the digestive tract in allergic patients therefore leads to an increased intestinal absorption of macromolecules but also to cromoglycate passage, which inhibits allergic reactions.

Using an animal model of gastric ulceration, in vivo and in vitro methods were developed to assess the importance of histamine in the cell turnover abnormalities previously found to occur in the preulcerous phase of gastric ulceration induced by mucosal anaphylaxis, and by using specific H1- and H2-receptor agonists and antagonists to define which receptors were involved. In ovalbumin-immunized animals, intramucosal ovalbumin injection led to a highly significant increase in cell turnover as measured by [3H]thymidine uptake (p less than 0.001). This increase was not affected by treatment with the H2 antagonist cimetidine, but was significantly reduced by the H1 antagonists promethazine and clemastine (p less than 0.01). A cell culture method was then used to examine the effects of added histamine and its antagonists. Histamine at a concentration of 10(-7) M was found to have a significant stimulatory effect on cell proliferation (p less than 0.001), and this effect was blocked by promethazine and clemastine but not by cimetidine. In contrast, histamine at a concentration of 10(-2) M had an inhibitory effect on cell proliferation which was accentuated by cimetidine and partially reversed by promethazine and clemastine. The H1 agonist betahistine was found to have stimulatory effects similar to those of histamine in low concentration (10(-7) M and 10(-6) M), but the H2 agonist 4-methyl histamine did not affect thymidine uptake at these concentrations. Both H1 and H2 agonists had a similar inhibitory effect at a concentration of 10(-2) M. It is concluded that histamine probably plays an important part in the changes in cell kinetics associated with gastric ulceration induced by this method, and that the trophic effects of histamine are mediated by H1-receptors. H2-receptors appear to have only a limited role in the control of cell proliferation.

IgA-, IgM-, IgG- and IgE-positive plasmocytes and mast cells were counted in the fundic mucosa of two groups of rats which underwent gastroenteroanastomosis or sham operation 12 months previously. Experimentally induced duodenogastric reflux constitutes a model for biliary gastritis which is characterized by a decrease in the IgA plasmocyte count along with an increase in IgG and IgE plasmocytes and, especially, an increase in mast cells. It is suggested that this allergic-type reaction is responsible for the hyperplasia of gastric crypts seen in biliary gastritis.