Spleen Tissue Of Mice Research Articles

Risk of Developing Hepatocellular Carcinoma following Depressive Disorder Based on the Expression Level of Oatp2a1 and Oatp2b1.

Background Accumulating evidence from prospective epidemiological studies has showed that depression disorder (DD) is a risk factor for cancer. The aim of this study is to explore the association of DD and the overall occurrence risk of hepatocellular carcinoma (HCC) and the mechanism. Methods In this study, 60 mice were randomly divided into four groups: Control group, DD group, HCC group, HCC-DD group. Mice received a chronic dose of reserpine to establish depression model, followed by Diethylnitrosamine and Carbon tetrachloride administration to establish HCC models. Behavioral depression was assessed by sucrose preference test (SPT) and the expression of Serotonin 1A (5-HT1A) receptor in the hippocampal. The expression of Oatp2a1 and Oatp2b1 in the digestive system tissues was detected by PCR and western blotting. Results Reserpine-administrated mice had a reducing sucrose preference at Day 14 compared with blank mice (P<0.05). The expression of 5-HT1A receptor in the hippocampal was decreased in DD mice compared with blank mice. The survival analysis indicated that the HCC mice with DD have poorer survival rate compared with the HCC mice. Compared with HCC mice, the expression of Oatp2a1 and Oatp2b1 was lower in liver and stomach tissue and higher in hepatic carcinoma and colon tissue of HCC-DD mice (P<0.05), and the expression of Oatp2a1 was higher in the spleen tissue of HCC-DD mice while the expression of Oatp2b1 was lower (P<0.05). However, no difference was found in the expression of Oatp2a1 and Oatp2b1 in the small intestine tissue between HCC group and HCC-DD group. Conclusions DD was the adverse factors for the overall occurrence risk of HCC. Mechanistically, be the downregulation of Oatp2a1 and Oatp2b1 in liver tissue induced by DD might be involved.

Differential tissue expression of erythroblast macrophage protein in a MRL/lpr mouse model of lupus.

The objective of this study was to observe the expression features of erythroblast macrophage protein (EMP) between the tissues of MRL/lpr mice, a mouse model of systemic lupus erythematosus (SLE), and control mice. We examined the serum ANA in both mice groups through indirect immunofluorescence (IIF). Expression features of EMP in bone marrow, liver, renal, spleen, brain, and lung tissues of the MRL/lpr mice and control mice groups were followed using quantitative real-time polymerase chain reaction (Q-PCR). Meanwhile, the expression of EMP was located through immunohistochemical (IHC) studies and the expressive cell identified through double immunofluorescent labeling. IIF showed that lupus mice have strong positive fluorescence, but no significant fluorescence was observed in control mice. Q-PCR detection revealed that EMP was expressed in the marrow, liver, renal, spleen, lung, and brain tissues of lupus mice. The highest levels were observed in the bone marrow, but there was no statistical difference between these tissues. EMP mRNA expression in the liver (t = 2.747, p = 0.01) and bone marrow (t = 3.853, p = 0.008) of lupus mice was significantly higher than in the control mice. However, no differences in EMP mRNA expression were observed in the renal, spleen, lung, and brain tissues between the lupus and control mice (p > 0.05). In addition, the IHC results showed that EMP protein is ubiquitously expressed in all of the tissues of the lupus and control mice. The positive expression rate in the bone marrow and liver tissues of the lupus mice was higher than in the control mice, but without an obvious difference in the other tissues. The double IF staining method shows that EMP protein was expressed in macrophages in the tissues of the lupus mice and the control mice. Our data showed that EMP is ubiquitously expressed in macrophages at all of the tissues of the lupus and control mice. However, the expression of EMP in bone marrow and liver tissues of lupus mice was higher than in the control mice, which indicates that EMP may be important in the development of SLE.

Preventive effect of Lactobacillus plantarum KSFY02 isolated from naturally fermented yogurt from Xinjiang, China, on d-galactose–induced oxidative aging in mice

Yogurt from Xinjiang, China, is a traditional and naturally fermented food, and abundant microorganisms are produced during its fermentation process. In this study, we carried out in vivo animal experiments to explore the effect of a newly isolated lactic acid bacterial strain, Lactobacillus plantarum KSFY02 (LP-KSFY02), on oxidative aging. We used d-galactose to induce oxidative aging in mice and analyzed the serum and tissues of those mice using molecular biology detection methods. The results showed that LP-KSFY02 could inhibit the decreases in the thymic, cerebral, cardiac, liver, spleen, and kidney indices of mice caused by oxidative aging. The LP-KSFY02 strain increased activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and glutathione (GSH) and reduced levels of nitric oxide (NO) and malondialdehyde in the serum, liver, and spleen of the oxidative aging mice. Pathological observation demonstrated that LP-KSFY02 alleviated damage to the liver and spleen of oxidative aging mice. Quantitative PCR showed that LP-KSFY02 effectively upregulated mRNA expression of neuronal nitric oxide synthase (Nos1), endothelial nitric oxide synthase (Nos3), copper/zinc superoxide dismutase (Sod1), manganese superoxide dismutase (Sod2), catalase (Cat), heme oxygenase-1 (Hmox1), nuclear factor erythroid 2 related factor 2 (Nfe2l2), γ-glutamylcysteine synthetase (Gclm), and quinone oxidoreductase 1 (Nqo1) in mouse liver and spleen and downregulated expression of inducible nitric oxide synthase (Nos2). Western blot analysis revealed that LP-KSFY02 effectively upregulated protein expression of SOD1, SOD2, CAT, GSH1, and GSH2 in mouse liver and spleen tissues. Therefore, LP-KSFY02 can effectively prevent d-galactose-induced oxidative aging in mice. Its efficacy was superior to that of Lactobacillus delbrueckii ssp. bulgaricus (LDSB) and vitamin C, which are commonly used in the medical field as antioxidants. Thus, LP-KSFY02 is a high-quality strain with probiotic potential.

Effect of ginkgo biloba L. extract on apoptosis and NF-κB signaling pathways in the spleen tissue of mice with radiation damage

Objective To investigate the influence of Ginkgo biloba L. extract (EGB) on the expression of apoptosis-related protein and NF-κB signaling pathway in the spleen tissue of mice with radiation damage. Methods The mice were divided randomly into normal control group (NC), irradiation control group (IC), low dose EGB group (IC+EGBL), medium dose EGB group (IC+EGBM) and high dose EGB group (IC+EGBH) according to the random number table, with 12 rats in each group. The mice in the low, middle and high dose EGB groups were given EGB respectively by 5, 10, 20 mg/kg, and the normal and irradiation control group were given saline by intraperitoneal injection once daily for 14 days. On the 15th day, the mice in all groups were uniformly irradiated with 4.0 Gy γ-rays for one time except normal control group. After 24 hours, Bcl-2, Bax and caspase-3 protein expression were measured by immunohistochemical method. The IKKβ expression was detected by qRT-PCR method, and the content of NF-κB p65 and IKKα in serum was detected by Elisa method in spleen tissue. Results Comparing with IC group, the expression of Bax (54.31 ± 1.59, 42.04 ± 1.56, 32.08 ± 2.43 vs. 68.68 ± 3.12) and caspase-3 protein (55.73 ± 2.61, 45.81 ± 2.59, 36.78 ± 2.23 vs. 72.18 ± 1.84) in IC+EGBL, IC+EGBM and IC+EGBH group were significantly decreased (P<0.05), the expression of Bcl-2 protein (30.33 ± 1.28, 39.80 ± 2.86, 44.42 ± 3.64 vs. 22.80 ± 2.01) in IC+EGBL, IC+EGBM and IC+EGBH group significantly increased (P<0.05), the expression of IKKβ mRNA (1.43 ± 0.06, 1.31 ± 0.06, 1.17 ± 0.09 vs. 1.64 ± 0.10) and the level of NF-κB p65 (129.38 ± 8.41 pg/ml, 111.28 ± 9.09 pg/ml, 95.41 ± 6.88 pg/ml vs. 145.64 ± 6.29 pg/ml) and IKKα (160.10 ± 8.94 pg/ml, 144.00 ± 8.36 pg/ml, 108.84 ± 13.74 pg/ml vs. 176.38 ± 8.54 pg/ml) in IC+EGBL, IC+EGBM and IC+EGBH group significantly decreased (P<0.05). Conclusions The EGB can reduce the expression of apoptotic protein Bax and caspase-3 in spleen cells induced by radiation, elevate the expression level of apoptotic protein Bcl-2, and inhibit the damage caused by radiation by regulating NF-κB signaling pathway. Key words: Ginkgo folium; Extracts; Radiation protection; Apoptosis; NF-kappa B; Mice

Inhibition of Autophagy Alleviates Cadmium-Induced Mouse Spleen and Human B Cells Apoptosis.

Cadmium (Cd) is a toxic heavy metal that can accumulate and cause severe damage to many organs, such as liver, kidney, lung, etc. Cd also significantly suppresses immunity, however, the underlying mechanism involved in Cd-induced immunnotoxicity is still unclear. The present study indicated that semichronic Cd exposure (7 days) induced apoptotic damage of mouse spleen. In human Ramos B cells, Cd exposure also induced apoptosis, which was dependent on Cd-induced vacuole membrane protein 1 (VMP1) expression and autophagy. Cd-induced autophagy and apoptosis were abated when VMP1 expression was knockdown. In addition, Cd-induced VMP1 expression, autophagy, and apoptosis were dependent on the elevation of Ca2+ and reactive oxygen species (ROS). More important, Cd exposure also induced VMP1 expression and autophagy in mouse spleen tissue, and the intraperitoneal injection of the autophagy inhibitor chloroquine (CQ) into mice effectively reduced Cd-induced spleen apoptotic damage. Taken together, these results indicate Cd-induced autophagy, promotes apoptosis in immune cells, and inhibition of autophagy can alleviate Cd-induced spleen and immune cell apoptosis. This study might provide the groundwork for future studies on Cd-induced immunomodulatory effects and immune diseases.

Lactobacillus plantarum CQPC11 Isolated from Sichuan Pickled Cabbages Antagonizes d-galactose-Induced Oxidation and Aging in Mice.

Chinese pickled cabbage is a traditional fermented food that contains abundant microbes produced during the process of fermentation. In this work, an in vivo animal study was conducted to investigate the effects of a newly isolated lactic acid bacterium (Lactobacillus plantarum CQPC11, LP-CQPC11) on d-galactose-induced oxidation and aging in mice. Analysis of the serum and tissue samples of these mice using molecular biology approaches showed that LP-CQPC11 suppressed the decrease in thymus, brain, heart, liver, spleen, and kidney indices caused by oxidation and aging. Furthermore, LP-CQPC11 increased the levels of SOD (superoxide dismutase), GSH-Px (glutathione peroxidase), and GSH (glutathione), whereas it reduced the levels of NO (nitric oxide) and MDA (malondialdehyde) in the serum, liver, and spleen of oxidation and aging mouse models. Pathological observation indicated that LP-CQPC11 alleviated the damage caused by oxidation and aging on the liver and spleen of mice. qPCR analysis indicated that LP-CQPC11 effectively upregulated the expression of nNOS (neuronal nitric oxide synthase), eNOS (endothelial nitric oxide synthase), Cu/Zn-SOD (cuprozinc-superoxide dismutase), Mn-SOD (manganese superoxide dismutase), CAT (catalase), HO-1 (heme oxygenase-1), Nrf2 (nuclear factor-erythroid 2 related factor 2), γ-GCS (γ-glutamylcysteine synthetase), and NQO1 (NAD(P)H dehydrogenase [quinone] 1), but downregulated the expression of iNOS (inducible nitric oxide synthase) in the mouse liver and spleen. Western blot analysis showed that LP-CQPC11 effectively upregulated SOD1 (Cu/Zn-SOD), SOD2 (Mn-SOD), CAT, GSH1 (c-glutamylcysteine synthetase), and GSH2 (glutathione synthetase) protein expression in mouse liver and spleen tissues. These findings suggest that LP-CQPC11 can effectively prevent d-galactose-induced oxidation and aging in mice, and the effect is even better than that of the commonly used Lactobacillus delbruechii subsp. bulgaricus (LDSB) and vitamin C in the industry. Thus, LP-CQPC11 may be potentially employed as a probiotic strain.

Beneficial Effects of N-Acetyl-L-cysteine or Taurine Pre- or Post-treatments in the Heart, Spleen, Lung, and Testis of Hexavalent Chromium-Exposed Mice.

Hexavalent chromium[Cr(VI)] compounds may induce toxic effects, possibly via reactive intermediates and radicals formed during Cr(VI) reduction. In this study, we probed the possible effects of N-acetyl-L-cysteine (NAC) and taurine pre- or post-treatments on Cr(VI)-induced changes in lipid peroxidation and nonprotein thiols (NPSH) in mice heart, lung, spleen, and testis tissues. The mice were randomly assigned to six groups, consisting of control, Cr(VI)-exposed (20 mg Cr/kg, intraperitoneal ,ip), NAC (200mg/kg, ip) as pre-treatment and post-treatment, and taurine (1g/kg, ip) pre-treatment and post-treatment groups. Lipid peroxidation and NPSH levels were determined and the results were compared with regard to tissue- and antioxidant-specific basis. Exposure to Cr(VI) significantly increased lipid peroxidation in all tissues as compared to the control (p < 0.05); and consistent with this data, NPSH levels were significantly decreased (p < 0.05). Notably, administration of NAC and taurine, either before or after Cr(VI) exposure, was able to ameliorate the lipid peroxidation (p < 0.05) in all tissues. In the case of NPSH content, while the decline could be alleviated by both NAC and taurine pre- and post-treatments in the spleen, diverging results were obtained in other tissues. The effects of Cr(VI) on the lung thiols were abolished by pre-treatment with NAC and taurine; however, post-treatments could not exert significant effect. While thiol depletion in the heart was totally replenished by NAC and taurine administrations, NAC pre-treatment was partially more effective than post-treatment. In contrast with lipid peroxidation data, NAC treatment could not provide a statistically significant beneficial effect on NPSH content of the testis, whereas the effect in this tissue by taurine was profound. Thus, these data highlight the importance of tissue-specific factors and the critical role of administration time. Overall, our data suggest that NAC and taurine may have potential in prevention of Cr(VI)-induced toxicity in the heart, lung, spleen, and testis tissues.

The metabolism and distribution of imidazole alkaloids from Lepidium meyenii (Maca) in mouse by matrix-assisted laser desorption/ionization mass spectrometry imaging

In this paper, matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was visualized to investigate the metabolism and distribution of Lepidiline A (1,3-dibenzyl-2,4,5-trimethylimidazolium chloride) and Lepidiline B (1,3-dibenzyl-4,5-dimethylimidazolium chloride) from Lepidium meyenii (Maca) in heart, liver, spleen and kidney tissues of mice. The results show that the main metabolic pathways of two imidazole alkaloids in heart were the same as those in liver and kidney of mice, however, there were few metabolic distributions in spleen tissues. MALDI-MSI distribution of imidazole alkaloids in tissues at 3 h after oral Maca extract was clearly observed by high resolution mass spectrometry. Two imidazole alkaloids were uniformly distributed in heart, cortex of liver and medulla of kidney, but not in medulla of liver and cortex of kidney. This label-free imaging technique in situ will contribute to studying pharmacological mechanism of imidazole alkaloids.

Quantification of Toxoplasma gondii in the tissues of BALB/c mice after immunization with nanoliposomal excretory-secretory antigens using Real-Time PCR.

Toxoplasmosis is an infectious disease caused by the intracellular parasite Toxoplasma gondii. Although almost 1/3 of the world's population are seropositive, there is no effective vaccine against toxoplasmosis. Therefore, the development of an effective vaccine for control of toxoplasmosis is one of major concerns in parasitology. The aim of this study was to evaluate the efficacy of nano-liposomal excretory-secretory antigens (NLESA) in BALB/c mice. Excretory-secretory antigens (ESA) was obtained from tachyzoites, encapsulated in the liposome and studied by scanning electron microscope. BALB/c mice were immunized with NLESA and ESA, sterile phosphate-buffered saline (PBS). Immunization was performed three times at 14-day intervals and challenged with 1 × 104 tachyzoites of T. gondii RH strain four weeks later. The parasite load of mice blood, brain and spleen tissues were determined using quantitative PCR targeted at the repeated element (RE) gene. The immunization with NLESA and ESA induced a significant increase of anti-Toxoplasma IgG antibody compared with PBS group (P < 0.05). After challenge with tachyzoites, qPCR analyses showed significant reduction of parasite load in NLESA and ESA immunized mice compared with control group (P < 0.05). Also, NLESAs were more effective than ESAs and showed significantly reduced parasite load in blood (P = 0.001) and brain tissue (P = 0.01). The vaccination with NLESA showed more promising results comparing to ESA. Further studies are recommended in order to achieve effectiveness of the vaccine against T. gondii.

Immune Responses Induced by HSP60 DNA Vaccine against Toxoplasma gondii Infection in Kunming Mice.

Toxoplasma gondii can infect all the vertebrates including human, and leads to serious toxoplasmosis and considerable veterinary problems. T. gondii heat shock protein 60 (HSP60) is associated with the activation of antigen presenting cells by inducing initial immune responses and releasing inflammatory cytokines. It might be a potential DNA vaccine candidate for this parasite. A pVAX-HSP60 DNA vaccine was constructed and immune responses was evaluated in Kunming mice in this study. Our data indicated that the innate and adaptive immune responses was elicited by successive immunizations with pVAX-HSP60 DNA, showing apparent increases of CD3e+CD4+ and CD3e+CD8a+ T cells in spleen tissues of the HSP60 DNA-immunized mice (24.70±1.23% and 10.90±0.89%, P<0.05) and higher levels of specific antibodies in sera. Furthermore, the survival period of the immunized mice (10.53±4.78 day) were significantly prolonged during the acute T. gondii infection. Decrease of brain cysts was significant in the experimental group during the chronic infection (P<0.01). Taken together, TgHSP60 DNA can be as a vaccine candidate to prevent the acute and chronic T. gondii infections.

Gambaran Patologi Infeksi Trypanosoma evansi pada Mencit Pascapengobatan dengan Esktrak Ethanol Daun Kipahit (Tithonia diversifolia) (PATHOLOGIC FEATURES OF TRYPANOSOMA EVANSI INFECTION AFTER BEING TREATED WITH ETHANOLIC EXTRACT OF KIPAHIT LEAVES

Trypanosoma evansi is a causative agent of Surra disease, which can cause death on infected animals. Surra disease treatment using commercial drugs are expensive, so that it is necessary to look for an alternative drug based on herbal medicine. The present study was carried out to investigate the pathological feature of infected mice after being treated with ethanolic extract of kipahit leaves (Tithonia diversifolia). Forty two male mice weighted 25-30 g of DDY strain were used and divided into seven groups Each group consisted of six mice as replication i.e. uninfected mice as a negative control (P 0), mice infected 104/0.3 mL T. evansi intra peritoneally as a positive control (P 1), uninfected mice treated orally by ethanol extract of kipahit 25 mg/kg BW(P2), infected mice treated orally by ethanol extract of kipahit 25 mg/kg BW then gave booster (P2) on day 6 post infection – double doses (P3), infected mice treated orally by ethanol extract of kipahit 25 mg/kg BW – single dose (P4), infected mice treated by commercial drug – single dose (P5) and infected mice treated by commercial drug – double doses (P6). Anatomy pathological lessions such as splenomegaly, hepatomegaly and pailness of kidney could be identified in mice organs. The histopathologycal lessions such as edema, hemorhhage, infiltration of inflamatory cells and trypanosome parasites in blood vessels could found in liver, kidney, lung, cardiac, spleen tissues of mice, edema also vasculitis found in cerebrum. It was concluded that intraperitoneal administration kipahit ethanol extract at dose 25 mg/BW might potentially reduce the number of T. evansi in the bloodstream of infected animals, but could not prevent and recover from T. evansi inducing organs damage.&#x0D;

The role of CD39-expressing regulatory T cells following treatment with αCD52: an effective therapeutic against central nervous system autoimmunity

Abstract Alemtuzumab (αCD52), a novel immunotherapy approved for the treatment of relapsing multiple sclerosis (MS), induces the widescale deletion of lymphocytes, including self-reactive T cells. Using the mouse model of MS, experimental autoimmune encephalomyelitis (EAE), we previously found that following reconstitution of the immune compartment, there was an increased frequency of CD39+Foxp3+ regulatory T cells (Tregs) in the spleen and gut-associated lymphoid tissues of αCD52-treated mice. We hypothesized that the CD39-expressing Tregs that emerge following lymphocyte reconstitution would be suppressive against central nervous system autoimmunity. However, when CD39+ Tregs were purified from αCD52-treated mice and adoptively transferred to naïve recipients, they were unable to reduce the severity of EAE disease. We next used Foxp3/GFP reporter mice treated with αCD52 to compare the suppressive capacity of CD39+Foxp3+ versus CD39+Foxp3− Treg subsets and found that CD39+Foxp3+ Tregs, but not CD39+Foxp3− Tregs, were able to significantly reduce the proliferation of activated naïve T cells in vitro. Finally, we sought to determine if expression of CD39 was required for the therapeutic effects of αCD52 in the EAE model. Preliminary studies using a limited number of CD39 KO mice indicated that treatment with αCD52 was equally effective in both control and KO mice. Thus, our studies have identified potential differences in individual subsets of Tregs emerging after αCD52 treatment and suggest that expression of CD39 is dispensable for the short-term effectiveness of αCD52 in mice. Future studies are required to determine if CD39 expression on Tregs contributes to the long-term suppression of MS disease in patients treated with αCD52.

Dissemination of Orientia tsutsugamushi, a Causative Agent of Scrub Typhus, and Immunological Responses in the Humanized DRAGA Mouse.

Scrub typhus is caused by Orientia tsutsugamushi, an obligated intracellular bacterium that affects over one million people per year. Several mouse models have been used to study its pathogenesis, disease immunology, and for testing vaccine candidates. However, due to the intrinsic differences between the immune systems in mouse and human, these mouse models could not faithfully mimic the pathology and immunological responses developed by human patients, limiting their value in both basic and translational studies. In this study, we have tested for the first time, a new humanized mouse model through footpad inoculation of O. tsutsugamushi in DRAGA (HLA-A2.HLA-DR4.Rag1KO.IL2RγcKO.NOD) mice with their human immune system reconstituted by infusion of HLA-matched human hematopoietic stem cells from umbilical cord blood. Upon infection, Orientia disseminated into various organs of DRAGA mice resulted in lethality in a dose-dependent manner, while all C3H/HeJ mice infected by the same route survived. Tissue-specific lesions associated with inflammation and/or necroses were observed in multiple organs of infected DRAGA mice. Consistent with the intracellular nature of Orientia, strong Th1, but subdued Th2 responses were elicited as reflected by the human cytokine profiles in sera from infected mice. Interestingly, the percentage of both activated and regulatory (CD4+FOXP3+) human T cells were elevated in spleen tissues of infected mice. After immunization with irradiated whole cell Orientia, humanized DRAGA mice showed a significant activation of human T cells as evidenced by increased number of human CD4+ and CD8+ T cells. Specific human IgM and IgG antibodies were developed after repetitive immunization. The humanized DRAGA mouse model represents a new pre-clinical model for studying Orientia-human interactions and also for testing vaccines and novel therapeutics for scrub typhus.

Protection of Spleen Tissue of γ-ray Irradiated Mice against Immunosuppressive and Oxidative Effects of Radiation by Adenosine 5'-Monophosphate.

The immune system is very sensitive to radiation. This study revealed that adenosine 5′-monophosphate (5′-AMP) increased the DNA contents of the spleen and the spleen index of irradiated mice. Moreover, the exogenous 5′-AMP could significantly repair the ultra-structure of the damaged spleen through transmission electron microscopy. When indicators of the mouse immune system were assessed, the flow cytometry and enzyme-linked immunosorbent assay (ELISA) revealed that the administration of exogenous 5′-AMP could reduce the apoptosis in the splenic cells. It could also regulate the transition of cells towards S phase, increase the proportion of CD4+ and CD8+ cellular subsets, and enhance the secretion of interleukin-2 (IL-2), IL-4, IL-10, and interferon-γ (IFN-γ). These effects were associated with a decrease in oxidative stress, as evidenced by changes in superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), reduced glutathione (GSH), and malondialdehyde (MDA) levels of spleen tissues. These results suggested that exogenous 5′-AMP could repair the damaged spleen, increase the spleen index, and regulate the cell cycles and apoptosis. There was an increase in the production of various cytokines and play a protective role on the immune system of irradiated mice by dynamically adjusting the REDOX balance.

1,25(OH)2 D3 inhibited Th17 cells differentiation via regulating the NF-κB activity and expression of IL-17.

The role of vitamin D (VD) in innate and adaptive immune responses to tuberculosis is still unclear. Our research was aimed to uncover the effect of VD on Th17 cells and elucidate potential molecular mechanism. VDR-deficient and wild-type mice were used to obtain CD4 T cells. Th17 cells were induced and activated by Bacillus Calmette Guerin. Flow cytometry was used to analyse the apoptosis rate and degree of differentiation of Th17 cells in the treatment of 1,25(OH)2 D3 . The interaction between P65 and Rorc was determined by immunofluorescence assay, luciferase reporter assay, EMSA-Super-shelf assay and ChIP assay. Co-IP assay was carried out to test the interaction between VDR and NF-κB family proteins. qRT-PCR and Western blot were also performed to detect the levels of P65, RORγt and IL-17. The Th17 cells differentiation was suppressed by 1,25(OH)2 D3 in vitro. We confirmed that Rorc was a downstream gene of the transcription factor P65. VDR interacts with P105/P50, P100/P52 and P65 NF-κB family proteins. 1,25(OH)2 D3 inhibited the expression of RORγt/IL-17 by suppressing p65 transcription factor translocating to nucleus. In vivo experiments, the expression of IL-17 and RANKL was suppressed by 1,25(OH)2 D3 by VD receptor. Moreover, 1,25(OH)2 D3 suppressed the inflammatory infiltrates and inhibited the expression of P65, RORγt and IL-17 in the spleen tissues of model mice. Together, 1,25(OH)2 D3 suppressed the differentiation of Th17 cells via regulating the NF-κB activity.

Alternative splicing variant of the scaffold protein APPL1 suppresses hepatic adiponectin signaling and function

Adiponectin is an adipocyte-derived hormone with antidiabetic activities that include increasing the sensitivity of cells to insulin. Adaptor protein containing pleckstrin homology domain, phosphotyrosine-binding domain, and leucine zipper motif (APPL1) stimulates adiponectin signaling and promotes adiponectin's insulin-sensitizing effects by binding to two adiponectin receptors, AdipoR1 and AdipoR2, and the insulin receptor. In this study, we report an alternative splicing variant of APPL1 (APPL1sv) that is highly expressed in mouse liver, pancreas, and spleen tissues. The expression levels of APPL1sv in liver tissues were enhanced in a mouse model of obesity and diabetic dyslipidemia (i.e. db/db mice) and reduced in calorie-restricted mice compared with ad libitum-fed mice. APPL1sv overexpression or suppression inhibited or enhanced, respectively, adiponectin-stimulated phosphorylation of AMP protein kinase (AMPK) in mouse hepatocytes. We also found that APPL1sv binds to AdipoR1 and AdipoR2 under basal conditions and that adiponectin treatment reduces this binding. Overexpression of APPL1sv blocked adiponectin-induced interactions of APPL1 with the adiponectin receptors. Moreover, adenovirus-mediated and short hairpin RNA-based suppression of APPL1sv greatly reduced high fat diet-induced insulin resistance and hepatic glucose production in mice. Our study identifies a key suppressor of hepatic adiponectin signaling and insulin sensitivity, a finding that may shed light on identifying effective therapeutic targets for treating insulin resistance and type 2 diabetes.

N-3 polyunsaturated fatty acids restore Th17 and Treg balance in collagen antibody-induced arthritis.

N-3 polyunsaturated fatty acids (PUFA) have anti-inflammatory effects and were considered useful for the treatment of rheumatoid arthritis (RA). Recently, several studies suggested that n-3 PUFAs attenuated arthritis in animal model and human, however the mechanism is still unclear. Interleukin 17 (IL-17) is a pro-inflammatory cytokine mainly produced by T helper 17 (Th17) cells which cause tissue inflammation and bone erosion leading to joint destruction. In contrast, regulatory T (Treg) cells down-regulate various immune responses by suppression of naïve T cells. The imbalance between Th17 cells and Tregs cell is important for the pathogenesis of RA. Here, we investigated whether n-3 PUFAs attenuate arthritis in collagen antibody-induced arthritis (CAIA) model. We used fat-1 transgenic mice expressing the Caenorhabditis elegans fat-1 gene encoding an n-3 fatty acid desaturase that converts n-6 to n-3 fatty acids, leading to abundant n-3 fatty acids without the need of a dietary n-3 supply. Clinical arthritis score was significantly attenuated in fat-1 mice compared to wild type (WT) mice on day 7 (1.6±1.8, p = 0.012) and day 9 (1.5±1.6, p = 0.003). Ankle thickness also decreased significantly in fat-1 mice compared to WT mice (1.82±0.11, p = 0.008). The pathologic finding showed that inflammatory cell infiltration and bone destruction were reduced in fat-1 mice compared to WT. The expression levels of IL-17 and related cytokines including IL-6 and IL-23 decreased in the spleen and ankle joint tissue of fat-1 mice compared to WT mice. Furthermore, Treg cells were expanded in the spleen of fat-1 mice and Treg cell differentiation was significantly higher in fat-1 mice than in wild type (p = 0.038). These data suggest that n-3 PUFAs could attenuate arthritis through increasing the expression of FoxP3 and the differentiation of Treg, while reducing IL-17 production. Therefore, dietary supplementation of n-3 PUFAs could have a therapeutic potential for the treatment of RA.

Mitochondrial DNA deletions in tissues of mice after ionizing radiation exposure

Purpose: The objective of the study was to estimate the presence of large mtDNA deletions in brain and spleen tissues of mice four months after exposure to 2 and 5 Gy.Materials and methods: The male BALB/c mice underwent X-ray total-body acute radiation. Four months after irradiation, the mice were decapitated, and the samples of spleen and brain tissues were examined. A long-distance PCR was used to detect mtDNA deletions and their levels in samples of brain and spleen tissues.Results: Four months after irradiation the levels of mtDNA deletions in the brain and spleen tissues were higher in animals exposed to 5 Gy than in animals at an irradiation dose of 2 Gy and in control mice. The levels of deletions in the mice brain tissues were higher 4 months than 1 month after X-ray exposure to both doses (2 and 5 Gy). In spleen tissues, a higher level of deletions was observed only at an irradiation dose of 5 Gy.Conclusions: Our data have shown that ionizing radiation induces an increase of mtDNA copies with deletions in tissues of mice four months after the post-irradiation period. The level of deletions depends on the animal age, type of tissue, irradiation dose and length of the post-irradiation period.

Galectin-3 impacts Cryptococcus neoformans infection through direct antifungal effects

Cryptococcus neoformans is an encapsulated fungal pathogen that causes cryptococcosis, which is a major opportunistic infection in immunosuppressed individuals. Mammalian β-galactoside-binding protein Galectin-3 (Gal-3) modulates the host innate and adaptive immunity, and plays significant roles during microbial infections including some fungal diseases. Here we show that this protein plays a role also in C. neoformans infection. We find augmented Gal-3 serum levels in human and experimental infections, as well as in spleen, lung, and brain tissues of infected mice. Gal-3-deficient mice are more susceptible to cryptococcosis than WT animals, as demonstrated by the higher fungal burden and lower animal survival. In vitro experiments show that Gal-3 inhibits fungal growth and exerts a direct lytic effect on C. neoformans extracellular vesicles (EVs). Our results indicate a direct role for Gal-3 in antifungal immunity whereby this molecule affects the outcome of C. neoformans infection by inhibiting fungal growth and reducing EV stability, which in turn could benefit the host.

Tick-Borne Transmission of Murine Gammaherpesvirus 68.

Herpesviruses are a large group of DNA viruses infecting mainly vertebrates. Murine gammaherpesvirus 68 (MHV68) is often used as a model in studies of the pathogenesis of clinically important human gammaherpesviruses such as Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus. This rodent virus appears to be geographically widespread; however, its natural transmission cycle is unknown. Following detection of MHV68 in field-collected ticks, including isolation of the virus from tick salivary glands and ovaries, we investigated whether MHV68 is a tick-borne virus. Uninfected Ixodes ricinus ticks were shown to acquire the virus by feeding on experimentally infected laboratory mice. The virus survived tick molting, and the molted ticks transmitted the virus to uninfected laboratory mice on which they subsequently fed. MHV68 was isolated from the tick salivary glands, consistent with transmission via tick saliva. The virus survived in ticks without loss of infectivity for at least 120 days, and subsequently was transmitted vertically from one tick generation to the next, surviving more than 500 days. Furthermore, the F1 generation (derived from F0 infected females) transmitted MHV68 to uninfected mice on which they fed, with MHV68 M3 gene transcripts detected in blood, lung, and spleen tissue of mice on which F1 nymphs and F1 adults engorged. These experimental data fulfill the transmission criteria that define an arthropod-borne virus (arbovirus), the largest biological group of viruses. Currently, African swine fever virus (ASFV) is the only DNA virus recognized as an arbovirus. Like ASFV, MHV68 showed evidence of pathogenesis in ticks. Previous studies have reported MHV68 in free-living ticks and in mammals commonly infested with I. ricinus, and neutralizing antibodies to MHV68 have been detected in large mammals (e.g., deer) including humans. Further studies are needed to determine if these reports are the result of tick-borne transmission of MHV68 in nature, and whether humans are at risk of infection.