Main Immune Organs Research Articles

Classical Swine Fever Virus Infection Induces Endoplasmic Reticulum Stress-Mediated Autophagy to Sustain Viral Replication in vivo and in vitro.

Endoplasmic reticulum (ER) stress-mediated autophagy plays significant roles in replication and pathogenesis of many animal viruses. However, the relationship between ER stress, autophagy, and viral replication during in vivo and in vitro infection of classical swine fever virus (CSFV) remains unclear. In this study, we established a pig model for CSFV infection and found that viral loads of CSFV differed in 10 kinds of infected organs, and that the degree of tissue lesions was to some extent positively correlated with CSFV replication in vivo. Next, we found that CSFV infection not only induced ER stress and subsequently activated three unfolded protein responses (UPR) pathways including protein kinase R-like ER kinase (PERK), inositol requiring enzyme 1 (IRE1), and activating transcription factor-6 (ATF-6) pathways, but also triggered complete autophagy in main immune organs and partial nonimmune organs exhibiting severer pathological injuries and higher viral loads. However, only the IRE1 pathway and no autophagy were activated in some other nonimmune organs with slighter pathologies and lower viral loads. These results indicate a potential link between CSFV-induced ER stress and autophagy, both of which are associated with the CSFV replication in vivo. We further performed in vitro experiments and found that CSFV infection activates the PERK and IRE1 pathways and autophagy in cultured porcine kidney cell lines (PK-15) and macrophage cell lines (3D4/2), and pharmacological regulation of ER stress remarkably changed autophagic activities induced by CSFV, suggesting that CSFV-induced autophagy can be mediated by ER stress possibly via the PERK and IRE1 pathway. Furthermore, treatment with ER stress regulators significantly altered copy numbers of NS5B genes, expression of Npro proteins, and viral titers in CSFV-infected cells or in cells treated with autophagy regulators prior to CSFV infection, suggesting the requirement of ER stress-mediated autophagy for CSFV replication in vitro. Collectively, our data demonstrate that CSFV induces ER stress-mediated autophagy to sustain its replication in vivo and in vitro, which may be one of the potential strategies exploited by CSFV for immune evasion. This finding will provide new insights into mechanisms of replication and pathogenesis of CSFV, and development of new strategies for controlling CSF.

Is There a Link between Basal Metabolic Rate, Spleen Volume and Hepatic Growth Factor Levels in Patients with Obesity-Related NAFLD?

Background: Recent pieces of research point to a link between basal metabolic rate (BMR) and non-alcoholic fatty liver disease (NAFLD) or hepatic steatosis (HS). The spleen in obese patients is associated with the cardiovascular system. Enlargement of the spleen is suggestive of nonalcoholic steatohepatitis (NASH). Patients with NASH present an increase in growth factor (HGF) as well as those with advanced heart failure. Interleukin-16 and interleukin-12p40 levels were found to correlate significantly with BMI, and waist circumference. Aim: We tried to find a relationship between BMR, spleen length and HGF. Methods: We analysed retrospective data from 80 obese patients with NAFLD. We evaluated indices of indirect calorimetry by the bioimpendance analysis; carotid intima-media thickness (IMT), spleen length (SLD) and HS by ultrasonography; serum HGF, IL-16, IL-12p40 and IL-6 concentrations by a magnetic bead-based multiplex immunoassays and the severity of NAFLD by BARD score > 2. Results: HGF levels of the obese were higher than those of controls, p < 0.001. At linear regression, BMR was foreseen by spleen length (p < 0.001), which was predicted by HGF (p = 0.04). BMR was predicted by IL-16 (p = 0.005), which predicted HGF, p = 0.034. Only fat mass, among other factors, predicted early atherosclerosis, p = 0.017; IL-12p40 did not predict IMT, HGF and BMR (p = 0.57, 0.09 and 0.59, respectively). The BARD score > 2 was negatively predicted by BMR and FFM (p =0.032 and 0.031, respectively), at the logistic regression. Interesting findings at the extended regression (mediation effect) were: IL-16 was likely causal in predicting BMR by HGF levels; HGF was influential in predicting BMR by SLD level. HS was predicted by SLD in males (p = 0.014), of advanced age (p < 0.001) and by BMR (p < 0.001). IL-6 concentrations, but not BMR were influential in the prediction of HS by SLD. Conclusion: These data reinforce the concept that the immune system is a sensor of the metabolic state, showing a link between HGF levels and BMR, which is mediated by IL-16 (cytokine inducing a cascade of inflammatory factors), and ascertaining the influential effect of the spleen, as main immune organ.

Histological and transcriptomic responses of two immune organs, the spleen and head kidney, in Nile tilapia (Oreochromis niloticus) to long-term hypersaline stress

Hyperosmotic stress can adversely affect fish immunity, but little is known about the histological and transcriptomic responses of immune organs in fish in a hyperosmotic environment. This study evaluated the effects of long-term hypersaline conditions (16‰) on the growth, histology and transcriptomics of the two main immune organs, the spleen and head kidney, in Nile tilapia Oreochromis niloticus relative to those reared in freshwater for eight weeks. No differences in weight gain and specific growth rate were found between fish reared under these two salinities. Hyperosmotic stress induced a congestive or enlarged spleen. Platelet- and coagulation-related gene expression was significantly decreased in tilapia at 16‰. The red cell distribution width and value of the mean corpuscular hemoglobin were significantly greater in fish at 16‰ salinity than in control fish in freshwater. A large volume of melano-macrophages in the spleen and pigment deposition in both the spleen and head kidney were observed in the histological sections in fish at 16‰ salinity. Transmission electron microscopic results showed abnormal macrophages with deposition granules in the spleen and head kidney and more neutrophils in the head kidney of fish at 16‰ than in control fish. In total, 772 and 502 genes were annotated for significantly different expression in the spleen and head kidney, respectively, and corresponded to five and one significantly changed immune system pathways, respectively. The complement pathway in the spleen was significantly down-regulated at 16‰. This study indicates that long-term exposure of Nile tilapia to a hyperosmotic environment can induce splenomegaly, reduce coagulation function, enhance phagocytic activity and down-regulate the complement pathway in the spleen. The spleen is a more sensitive organ for immune responses to chronic ambient salinity stress than the head kidney in Nile tilapia.

Efficacy of different antimycotoxicosis compounds on the main immune organs and humoral immune response of broiler chickens fed on aflatoxin and/or ochratoxin contaminated diet

The ameliorative effects of some different antimycotoxicosis compounds (AMCs) against the adverse effects induced by intoxication of dietary aflatoxin (AF) and/or ochratoxin (OT) on the immune system performance in broiler chicks were investigated.AF (23 ppb), OT (17 ppb) and mixed doses were fed and in association with antimycotoxin feed additives; product A (combination of Eubacterium BBSH797 and Trichosporon mycotoxinivoran, plant extract, algae extract) or product B (nano-clay, seaweed extract, yeast cell wall, diatomaceous earth) from 1 day old chicks to 21 day. The intoxicated chicks treated with AMCs; product C(combination of bacterial cell wall extract, oligosaccharides, L-form bacteria extract, group of mycotoxin biotransforming enzymes, organic acid and salts, hepatic and renal tonic and vitamins)or product D in drinking water (isomalto-oligosaccharide, micronized-mannan oligosacharide, lactobacillus extract and cholestyramine) were used in the drinking water of 2 weeks old chicks after appearance of signs. Results revealed that chicks intoxicated with AF and/or OT showed significant (P<0.05) decrease in the relative weights of the main immune organs and the antibody titers against vaccines of Newcastle disease (ND) and Infectious bursal diseases (IBD) viruses, in addition to significant (P<0.05) increase in the cumulative histopathological lesion scores of thymus, bursa of Fabracius and spleen. The adverse effects of AF and/or OT on the relative weights and the histopathological lesion scores of the tested immune organs and the antibody titers against ND and IBD vaccination were significantly (P<0.05) improved with dietary supplementation of AMCs products A or B. While the treatment of the intoxicated chicks with AMCs products C or D in the drinking water showed partially and temporarily improvement in the relative weights and the histopathological lesion scores of the tested immune organs and did not improve the negative effects of AF and/or OT on the antibody titers against ND and IBD vaccines.

Prevenirea şi managementul infecţiilor postsplenectomie în trauma lienală

Currently, the spleen is considered the main peripheral immune organ. Infections are the most important consequences of the absence of spleen and may evolve to severe sepsis (OPSI = Overwhelming Post Splenectomy Infection) and death. Therefore, in the context of trauma and not only, spleen conservation or autologous spleen implant are required to prevent postsplenectomy infections. In addition, patients require immunoprophylaxis (Pneumococcal Vaccination Haemophilus influenzae type b and Meningococcal vaccination) and chemoprophylaxis (penicillin V/amoxicillin).

Impact of selenium supplementation on fish antiviral responses: a whole transcriptomic analysis in rainbow trout (Oncorhynchus mykiss) fed supranutritional levels of Sel-Plex®.

BackgroundSelenium (Se) is required for the synthesis of proteins (selenoproteins) with essential biological functions. Selenoproteins have a crucial role in the maintenance of cellular redox homeostasis in nearly all tissues, and are also involved in thyroid hormone metabolism, inflammation and immunity. Several immune processes rely on Se status and can be compromised if this element is present below the required level. Previous work has supported the notion that when Se is delivered at levels above those deemed to be the minimal required but below toxic concentrations it can have a boosting effect on the organism’s immune response. Based on this concept Se-enriched supplements may represent a valuable resource for functional feeds in animal farming, including aquaculture.ResultsIn this study we tested the effects of Se supplemented as Sel-Plex during an immune challenge induced by polyinosinic:polycytidylic acid (poly(I:C)), a pathogen-associated molecular pattern (PAMP) that mimics viral infection. Trout were fed two diets enriched with 1 or 4 mg Se Kg−1 of feed (dry weight) by Sel-Plex addition and a commercial formulation as control. The whole trout transcriptomic response was investigated by microarray and gene ontology analysis, the latter carried out to highlight the biological processes that were influenced by Sel-Plex supplementation in the head kidney (HK) and liver, the main immune and metabolic organs in fish. Overall, Sel-Plex enrichement up to 4 mg Se Kg−1 induced an important response in the trout HK, eliciting an up-regulation of several genes involved in pathways connected with hematopoiesis and immunity. In contrast, a more constrained response was seen in the liver, with lipid metabolism being the main pathway altered by Se supplementation. Upon stimulation with poly(I:C), supplementation of 4 mg Se Kg−1 increased the expression of principal mediators of the antiviral defences, especially IFN-γ, and down-stream molecules involved in the cell-mediated immune response.ConclusionsSupplementation of diets with 4 mg Se Kg−1 using Sel-Plex remarkably improved the fish response to viral PAMP stimulation. Sel-Plex, being a highly bioavailable supplement of organic Se, might represent a suitable option for supplementation of fish feeds, to achieve the final aim of improving fish fitness and resistance against immune challenges.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-2418-7) contains supplementary material, which is available to authorized users.

Effects of dietary lipid sources on apoptotic and immune gene expression in head kidney of olive flounder (Paralichthys olivaceus)

It can be hypothesized that dietary fatty acids can modulate immune responses in fish by inducing apoptosis of immune cells since dietary polyunsaturated fatty acid (PUFA) increase apoptosis by oxygen radicals generated by peroxidation. Thus we examined the effects of deferent dietary lipid sources such as squid liver oil (FO), linseed oil (LO) and soybean oil (SO) on oxidation (Cytochrome C oxidase; COS), apoptosis (TNF-  Scinderin like) and immune (IL-1  and NKEF) gene expression in the main immune organ (head kidney) in olive flounder (Paralichthys olivaceus) by Q-PCR analysis after feeding diets containing each oil (5%) for 15 weeks. Linseed oil and soybean oil were chosen to compare n-3 or n-6 enriched vegetable oils, respectively. Consequently, COS, TNF-α and Scinderin like gene expression was increased in SO group, indicating the induction of oxidation and apoptosis. Meanwhile, no significant difference was found in immune gene expression. In conclusion vegetable oils containing n-3 PUFA like linseed oil seems to be more suitable lipid source than soybean oil for replacement of fish oil in flounder since n-6 PUFA in SO leads to activation of apoptosis pathways within the cellular damage in head kidney.

Role of the spleen in the development of steatohepatitis in high-fat-diet-induced obese rats

Obesity is considered a systemic low-grade inflammatory state. Although the spleen is the main immune organ with a close anatomical relationship with the liver, its role in the progression of fatty liver disease remains uncertain. Therefore, we sought to clarify the functional role of the spleen in the development of steatohepatitis in high-fat (HF)-diet-induced obese rats. Male Sprague-Dawley rats were fed HF food and divided into two groups, a splenectomy (SPX) group and a sham-operation (Sham) group. The liver and abdominal white adipose tissue (WAT) were removed one and six months after surgery, and we evaluated the effects of SPX on WAT and HF-induced fatty liver. SPX rats exhibited worse dyslipidemia and inflammatory changes in WAT one month after surgery. Hepatic steatosis and inflammation were accelerated by SPX, based on the time after surgery. At one month after surgery, the tissue triglyceride content increased in SPX rats, compared with Sham controls (P < 0.05). The liver histology also showed a worsening of steatosis in those rats. At six months after SPX, dramatic inflammatory and fibrotic changes were observed in liver tissue sections. Hepatic carnitine palmitoyltransferase-1 was suppressed at one and six months after SPX (P < 0.05 for each). WAT and liver tissue levels of inflammatory markers such as tumor necrosis factor-α, and the expression of Kupffer cells were all increased at six months in SPX rats, compared with Sham controls (P < 0.05 for each). Our results indicate that the preservation of the spleen contributes to the prevention of the progression of hepatic steatosis to steatohepatitis in obese rats.

Extracts of Lycoris aurea induce apoptosis in murine sarcoma S180 cells.

Lycoris species have been known since long ago as a multi-utility ethnomedicinal herbal in China. It has been reported to exhibit a number of properties such as anticancer, neuroprotective, and antibacterial activities. In the present study, the anticancer efficacy of dichloromethane extracts of Lycoris aurea (DELA), was evaluated both in vivo and in vitro using murine sarcoma 180 cells. To evaluate the effects of DELA on apoptotic cell death, flow cytometry and Western blotting were performed. DELA demonstrated promising inhibition effects on sarcoma 180 cells in vitro and a 53.49% inhibitory rate on cancer cells in vivo. DELA treatment increased thymus indices and spleen indices in vivo, indicating that it reduced tumours, but did not damage the main immune organs. The DELA-evoked increase in apoptotic cell death was accompanied by occurrence of cleaved caspase-3 and decreases in the ratio of Bcl-2/Bax. Further purification and LCMS analysis showed DELA contained homolycorine, 2α-hydroxyoduline, oduline, hippeastrine, 2α-hydroxy-6-O- methyloduline, and 2α-methoxy-6-O-methyloduline. These results indicate that DELA exerted its anticancer effects, at least in part, by inducing cancer cell apoptosis and thus can be considered as a potential candidate agent for treatment of cancer.

Transferrin gene expression in response to LPS challenge and heavy metal exposure in roughskin sculpin ( Trachidermus fasciatus)

Transferrin plays an important role in immune response of vertebrates. In the present study, a transferrin cDNA with a partial 5′ UTR of 7 bp and a complete 3′ UTR of 345 bp was obtained from the liver of roughskin sculpin, Trachidermus fasciatus, which encodes a deduced 681 amino acid protein containing an N-terminal signal peptide and two conserved lobes. In the N-terminal lobe, the anion-binding residue Arg was substituted with Lys, which represents a common feature in fish and implies a selective preference in the transferrin evolutionary process. In contrast to mammalian transferrin, the roughskin sculpin transferrin did not contain potential N-glycosylation sites, similar to those obtained in cyprinid fish, but not in salmonid fish. Quantitative real-time polymerase chain reaction demonstrated that the transferrin transcripts were abundant in the liver, but also significant in the brain, with a lesser expression in the other nine tissues. The temporal expression profiles were detected during the LPS challenge and heavy metal exposure experiment. Transferrin mRNA expression decreased in the liver in both experiments. Nevertheless, in the main immune organs (skin, blood, and spleen), transferrin mRNA expression was up-regulated significantly. These results suggest that transferrin is involved in the innate immune response of roughskin sculpin.

Laser Therapy of Irradiated Traumatized Skeletal Muscles and State of Immune System in Animals

Problems statement: This study was designed to examine the effect of laser therapy on regeneration of traumatized skeletal muscles and on dynamics of immune responses in thymus and bone marrow under ionizing radiation. Approach: After local X-ray radiation of both rat gastrocnemius muscles at a dose 20 Gy or after total γ-irradiation of animals at a semiletal dose 6 Gy, the muscles were damaged considerably as contractile organ (full cut across muscle). Then laser therapy was carried out with a helium-neon laser (632.8 nm) or infrared pulsed laser (890 nm), 10 treatments by 3 min during the first 15 or 30 days after trauma.The investigations were performed using histological, biochemical, cytological, cytogenetic and morphometric methods. Results: The received data showed that under ionizing radiation, He-Ne and IR laser irradiation stimulated the regeneration of skeletal muscles, improved the healing of skin-muscle wound and stimulated the prosesses of postradiation recovery in cells of the thymus and bone marrow. The regenerative activity of radiated traumatized muscles and the level of reduction of cytogenetic damages in bone marrow and the thymus varied depending on the period for which regenerative muscles were exposured to laser light, as well as the optical characteristics of lasers and the degree of radiation damages. The early appearance of reactive changes in structure of the thymus and the display them during 60 days point to increasing function of organ. Conclusion: Under local and total ionizing radiation and laser therapy of solely damaged skeletal muscles, positive dynamics of regeneration processes and improvement of cytological and cytogenetic indexes for main immune organs (the thymus and bone marrow) were observed. Herewith, the functional loading on thymus increased. We suggest that the findings should be considered in clinical practice.

Effect of Chito-oligosaccharide Supplementation on Immunity in Broiler Chickens

This study was conducted to determine the effects of dietary supplementation of either 100 mg/kg chito- oligosaccharide (COS) or chlortetracycline (CTC) with corn-soybean-fish meal on immunity in broiler chickens. A total of 147 one-day old male broiler chicks were randomly allocated to 3 treatments with 7 replicate pens per treatment and 7 birds per pen. The experimental diets consisted of a control diet based on corn, soybean and fish meal without COS and any antibiotic supplement and similar diets supplemented with either CTC (80 mg/kg from d 1 to 21 and 50 mg/kg from d 22 to 42) or COS (100 mg/kg from d 1 to 42). During the entire experimental period, all birds had ad libitum access to diets and water. The main immune organ indices, T- lymphocyte proliferation, serum cytokine concentrations, serum NO level and serum iNOS activity were measured on d 21 and d 42. On d 21, broilers fed 100 mg/kg COS had improved (p<0.01) indices of spleen, thymus, and bursa of Fabricius compared with the control and CTC birds. Birds receiving 100 mg/kg COS had higher (p<0.05) serum concentrations of IL-1β, IL-6, IgM, NO and iNOS than birds on the control treatment. Serum Ca 2+ level of birds fed 100 mg/kg COS tended to be higher (p = 0.049) than in birds fed CTC. On d 42, the birds fed 100 mg/kg COS had higher (p<0.05) concentrations of TNF-α and IgM in serum than birds in both the CTC and control treatments. Birds fed 100 mg/kg COS had a higher concentration of IFN-γ than the control group. In conclusion, dietary supplementation of COS appeared to improve the immunity of broilers by promoting the weight of the main immune organs, increasing IgM secretion, stimulating microphages to release TNF-α, IL-1β, IL-6 and IFN-γ, and activating iNOS to induce NO. (Key Words : Chito- oligosaccharide (CTC), Immune Organ Indices, Cytokines, NO, Broiler)

Modulation of Drug-Metabolizing Systems by Bacterial Endotoxin in Carp Liver and Immune Organs

This report describes a study of the effects of bacterial endotoxin [lipopolysaccharide (LPS)] on cytochrome P450 levels and ethoxyresorufinO-deethylase (EROD) and glutathioneS-transferase (GST) activities in liver and two main immune organs of carp: spleen and head kidney. Also studied was the paucity of the carp drug-metabolizing system in an environment subject to pollution by a polycyclic aromatic hydrocarbon, 3-methylcholanthrene (3MC), when fish respond to an immune activation by lipopolysaccharide (LPS). In the presence of bacterial endotoxin the basal cytochrome P450 levels were decreased in liver and spleen. EROD activity was increased in liver and basal GST activity was increased in spleen. When fish were treated concomitantly with 3MC and LPS, a suppression of cytochrome P450 induction in liver and head kidney was observed. EROD activity induced by 3MC was not modified by administration of LPS. GST activity was suppressed by treatment with LPS and inducing agent in liver and head kidney. In the present study it was found that endotoxin can have profound and differential effects on fish basal biotransformation of drugs in the liver and immune organs. Also, the induction of biotransformation enzymes by 3MC was modified when fish responded to an immune stimulation.

5. Locally advanced non-small cell lung cancer: The “local” issue

There exists maternal immunological modification in maternal immune organs during early pregnancy in mammals. Gonadotropin releasing hormone (GnRH) is widely distributed in vertebrate tissues, including immune organs. However, it is unclear that early pregnancy induces expression of GnRH and GnRH receptor (GnRHR) in ovine immune organs. The objective of this study was to explore the expression of GnRH and GnRHR in main immune organs (thymus, lymph node, spleen, and liver) during early pregnancy in sheep. Ovine thymus, lymph node, spleen and liver were sampled at day 16 of estrous cycle, and days 13, 16, and 25 of pregnancy. The expression of GnRH and GnRHR was detected through real-time quantitative PCR, Western blot and immunohistochemistry analysis. The results indicated that early pregnancy induced upregulation of mRNA and protein levels of GnRH and GnRHR in the maternal lymph node, spleen and liver, and mRNA and protein of GnRH in the maternal thymus, but mRNA and protein of GnRHR decreased in the maternal thymus during early pregnancy. In summary, the mRNA and protein levels of GnRH and GnRHR were changed in maternal thymus, lymph node, spleen and liver in a tissue specific manner during early pregnancy in sheep.

Immune Restoration in Children after Partial Splenectomy

Splenectomy (SE) is recognized to be a therapeutical approach in treating children with severe autoimmune diseases (chronic idiopathic thrombocytopenia; hemolytic anemia) or hypersplenism because of portal hypertension. Nevertheless, removal of a main immune organ results in elevated infection risk for these patients. Partial splenectomy (PSE) was developed as a therapeutical compromise to retain immunologically active spleen tissue. Here, we document the analysis of immune parameters obtained from children after both partial and total splenectomy, which have been followed up for a period of more than 6 years: (i) Lymphocytes from both groups of patients failed to produce IgG in response to pokeweed mitogen in vitro. This was observed in 11/20 splenectomized patients even 10 years after operation, whereas in PSE patients a restoration of this parameter after 1–2 years was seen. (ii) In patients after PSE, but not in splenectomized persons, an elevated number of HLA-class II positive cells had been detected suggesting a different situation of immune regulation following this operation. However, in parallel with an improvement of B cell in vitro activity this parameter was found to achieve normal values. Our findings indicate that partial splenectomy may be a therapeutical alternative, if the therapeutic goal can be achieved by this procedure.