Interleukin-2 mRNA Expression Research Articles

Evaluation of IL 12 gene expression and serum levels of OxPL / apoB in patients with coronary artery disease

Background: Atherosclerotic lesions are formed as a result of low-density lipoprotein (LDL) accumulation, which is produced by oxidizing enzymes and oxidized phospholipids (OxPL). Phospholipid oxide causes inflammation-inducing activation genes and hyper-inflammation in addition to the initiation of inflammation and expression of Th1 cytokines. Interleukin 12 (IL-12) is one of the most significant stimulants of inflammation and immune responses, among the Th1cytokines. Therefore, it may play a significant role in contributing to atherosclerosis. The quantitative expression of IL-12 mRNA, along with serum OxPL levels, has been analyzed in patients with coronary artery disease (CAD). Methods: Coronary angiography patients, aged 45 to 65 years and referred with chest pain, were examined. The patients were classified into normal coronary arteries (N = 38) and severe three-vessel involvement coronary arteries (N = 41). Molecular testing was performed using real-time PCR. Also, an ELISA test was used to check the OxPl level. Results: Expression of IL12 in Individuals with coronary artery disease was increased but was not statistically significant.

A Matched Case-Control Study of the Serum Level and Gene Expression of IL-6 and IL-12 in Patients with Schizophrenia Spectrum Disorder: An Azeri Acute Phase/Recent-Onset Psychosis Survey (ARAS) Study

Background: The causes of schizophrenia spectrum disorders are believed to be multifactorial, with both genetic and environmental factors, as well as gene-environment interactions, influencing the course of this disorder. Research suggests that immunological factors may play a role in the development of psychotic disorders. Objectives: The aim of this study is to evaluate and compare the serum levels of interleukin-6 (IL-6) and interleukin-12 (IL-12), as well as the expression levels of IL-6 and IL-12 genes, between an Iranian Azeri population of patients with first-episode psychosis and healthy controls. Methods: Forty patients who had recently experienced first-episode non-affective psychosis were allocated to the patient group. Forty healthy volunteers, matched for age and gender, were also recruited as controls. The study was conducted in 2020 in Tabriz, Iran. Peripheral blood samples were collected (using the Ficoll-Paque process) and assessed to determine the expression levels of IL-6 and IL-12 mRNA (using real-time Polymerase Chain Reaction-PCR- system), as well as the serum levels of IL-6 and IL-12 (with a sandwich ELISA kit). GraphPad Prism Version 6.0 was used for all statistical analyses. Results: Each group consisted of 40 participants in this study. Patients had significantly higher levels of IL-6 mRNA expression compared to the healthy controls (P < 0.0001), while no significant difference was found between the two groups in terms of IL-12 mRNA expression (P = 0.5697). A similar pattern was reported for the serum levels of IL-6 (P < 0.0001) and IL-12 (P = 0.0777). The area under the curve (AUC), which indicates the accuracy of the predictive model, was 0.9669 for the level of IL-6 mRNA expression (P < 0.0001). This Area (AUC) was 0.9584 for the serum concentration of IL-6 (P < 0.0001). The levels of IL-6 or IL-12 were not correlated with the severity of symptoms, gender, or age of the patients. Conclusions: The expression level of IL-6 mRNA and the serum concentration of IL-6 were altered in patients with first-episode non-affective psychosis. This finding supports the role of inflammatory pathways in the pathogenesis of psychosis. According to our results, measurement of the circulating level of IL-6 may be useful in distinguishing patients with first-episode psychosis from healthy individuals.

Dietary lysozyme and avilamycin modulate gut health, immunity, and growth rate in broilers

BackgroundAttempts to use dietary lysozyme (LYZ) as an alternative to antibiotics in broilers have been successful, but further research is needed for effective use. Here, we compared the differences between LYZ and avilamycin (AVI) feed additives for growth performance, gut health and immunity of broilers. One-day old, one hundred and twenty broiler chicks (Ross 308) were randomly allocated into three groups consisting forty birds in each group. Standard diet without supplementation was applied as the control group (I), while the chicks of the other groups were supplemented with 100 mg of AVI per kg diet (AVI, group II), and 90 mg LYZ per kg diet (LYZ, group III) for five consecutive weeks.ResultsBody weight, feed conversion ratio, body weight gain, and European production efficiency factor were markedly (p < 0.05) increased in both AVI and LYZ groups in relation to CON group, but the feed intake and protein efficiency ratio were not affected. Both AVI and LYZ significantly (p < 0.001) upregulated the mRNA expression of ileal interleukin-18 (IL-18), interferon-gamma (IFN-γ), and interleukin-10 (IL-10), interleukin-2 (IL-2), and glutathione peroxidase (GSH-PX) genes compared to CON group. However, IL-2, IL-10, IL-18, and GSH-PX genes were markedly (p < 0.01) upregulated in LYZ compared to the AVI group. LYZ treated group had a significant increase (p < 0.05) in the serological haemagglutination inhibition titers of H5N1 vaccination and a significant decrease (p < 0.0001) in coliform counts compared to control and AVI groups, but all growth parameters were nearly similar between AVI and LYZ groups. The VH and VH/CD were markedly higher in LYZ than AVI and control groups.ConclusionExogenous dietary lysozyme supplementation by a dose of 90 mg/kg broilers’ diet induced better effects on intestinal integrity, fecal bacterial counts, immune response, and growth performance which were comparable to avilamycin. Therefore, dietary lysozyme could safely replace avilamycin in the broiler chickens’ diet. However, further experimental studies regarding the use of lysozyme in commercial broilers, both in vitro and in vivo, targeting more communities of intestinal microbiome and explaining more details about its beneficial effects need to be conducted.

Interleukin-4 mRNA Expression in Laryngeal Squamous Cell Carcinoma Patients with or without Lymph Node Involvement

Background: Interleukin (IL)-4 is a member of T helper 2 (Th2) axis produced by T-lymphocyte and mast cell. It has been shown that IL-4 expression changes in tumor tissues. The main objective of this study is to investigate the expression of IL-4 mRNA in patients with Laryngeal Squamous Cell Carcinoma (LSCC) with or without lymph node involvement. Materials and Methods: mRNA expression of IL-4 in LSCC tissues were detected by quantative Real-Time PCR (qRT-PCR). Expression of IL-4 gene was compared between lymph node positive and negative patients with Squamous Cell Carcinoma of Larynx. Results: No statistically significant association was found in expression of IL-4 between lymph node positive and negative patients. Conclusion: It seems that IL-4 has no important effect on the involvement of lymph node in LSCC. However, to achieve a definite conclusion more investigations are certainly required.

Effects of rutin supplementation on intestinal morphology, antioxidant capacity, immunity, and gut microbiota of laying hens fed a diet containing stored soybean meal

Protein oxidation of soybean meal (SBM) during storage may have adverse effects on the intestinal health of laying hens. Moreover, rutin has anti-inflammatory and antioxidant properties which might be used as a feed additive to mitigate the intestinal damage caused by oxidised protein of SBM. This study aimed to investigate the effects of rutin supplementation on intestinal morphology, antioxidant capacity, immunity and caecal microbiota in laying hens fed a diet containing stored SBM. A total of 384 Hy-Line Brown laying hens (220 days) were randomly allocated into four groups with eight replicates of 12 laying hens each according to a 2 × 2 factorial arrangement with 2 types of SBM (FSM: SBM was stored in the cold storage warehouses at −20 °C for 45 days, and was considered as fresh and control SBM; RTSM: SBM was stored in room temperature warehouse (15 °C to 25 °C), average temperature was 20 °C for 45 days) and 2 levels of rutin (0 and 500 mg/kg). The results showed that the RTSM diet decreased the ileal glutathione peroxidase (GSH-Px) activity, the jejunal superoxide dismutase 2 (SOD2) and ileal NAD(P)H: quinone oxidoreductase 1 (NQO1) mRNA expression levels (p < 0.05), and tended to decrease the jejunal superoxide dismutase 1 (SOD1), ileal glutathione peroxidase 1 (GPX1) and increase the jejunal interleukin-1β (IL-1β) and interleukin-4 (IL-4) mRNA expression levels. Dietary rutin decreased the jejunal crypt depth (CD) (p < 0.05), increased the jejunal total antioxidant capacity (T-AOC) and GSH-Px activities (p < 0.05), decreased the content of interferon-γ (IFN-γ) in the jejunum (p < 0.05), and significantly reduced levels of immunoglobulin G (IgG), IL-1β, tumour necrosis factor-α (TNF-α), IFN-γ, and IL-4 in the ileal mucosa (p < 0.05). Dietary rutin increased SOD2, nuclear factor E2-related factor 2 (Nrf2) and NQO1 mRNA expression levels in the jejunum and GPX1, Nrf2 and NQO1 mRNA expression levels in the ileum, and decreased nuclear factor-κB (NF-κB), IL-1β, IFN-γ and IL-4 mRNA expression in the jejunum and NF-κB, IL-1β, TNF-α, IFN-γ and IL-4 mRNA expressions in the ileum. What’s more, dietary rutin changed the caecal microbiota. PCoA analysis indicated significant structural differences among four groups (p < 0.05), and SBM × Rutin interactions were found in Actinobacteriota and unclassifiedk_norank_d_ Bacteria at the phylum level (p < 0.05). These results suggested that RTSM had slight adverse effects on the intestinal health, and dietary rutin improved intestinal morphology, exerted antioxidant and anti-inflammatory effects via Nrf2 and NF-κB signal pathways, and changed the composition of caecal microbiota. Highlights The RTSM diet has adverse effects on the intestinal health of laying hens compared with the FSM diet. Dietary rutin improved the intestinal health by increasing the intestinal morphology, antioxidant capacity and anti-inflammatory effects. Dietary soybean meal and rutin changed the composition of caecal microbiota.

Resveratrol improves emamectin benzoate-induced pyroptosis and inflammation of Ctenopharyngodon idellus hepatic cells by alleviating oxidative stress/endoplasmic reticulum stress

Emamectin benzoate (EMB) is the most widely used pesticide in the world and contributes to water pollution. Owing to the lack of a specific antidote, EMB has a severe negative impact on the health of aquatic organisms. Resveratrol (RES), a substance with antioxidant capacity, is secreted by the fruits of many plants. This study was to explore the protection of RES against EMB-induced pyroptosis and inflammatory response in grass carp (Ctenopharyngodon idellus) hepatic liver (L8824) cells by oxidative stress/endoplasmic reticulum (ER) stress. The results showed that compared to the CON group, EMB induced oxidative stress in L8824 cells with the increase of reactive oxygen species (ROS), methane dicarboxylic aldehyde (MDA), and hydrogen peroxide (H2O2) contents and the decrease of total superoxide dismutase (t-sod) and glutathione peroxidase (gsh-px) activities (P < 0.05). In addition, EMB triggered ERS, increasing the relative mRNA expression of protein kinase R-like endoplasmic reticulum kinase (perk), inositol requiring enzyme 1 alpha (ire1α), glucose-regulated protein 78 (grp78), activating transcription factor 4 (atf4), activating transcription factor 6 (atf6), and CCAAT-enhancer-binding protein homologous protein (chop) and the protein expression of eukaryotic initiation factor 2α (eif2α), chop, atf6, and atf4. Meanwhile, EMB further induced pyroptosis by upregulating the mRNA and protein expression of nlrp3, aptamer protein (asc), caspase-1, gsdmd, interleukin-1β (il-1β), and interleukin-18 (il-18). EMB also induced inflammation in L8824 cells by increasing the mRNA expression of interleukin-2 (il-2), interleukin-6 (il-6), tumor necrosis factor-α (tnf-α), and ifn-γ and decreasing the content of interleukin-10 (il-10). However, compared to the EMB group, the oxidant indices and expression of genes related to ER stress, pyroptosis, and pro-inflammatory factors were significantly down-regulated (P < 0.05), whereas the antioxidant indicators and anti-inflammatory factor were significantly up-regulated in the EMB + RES group (P < 0.05). In conclusion, EMB caused hepatocytes pyroptosis and inflammation in grass carp, and RES could alleviate EMB-induced pyroptosis and inflammation in L8824 cells by ameliorating oxidative stress/ER stress.

The protective effects of silymarin nanoemulsion on 5-fluorouracil-induced gastrointestinal toxicity in rats

5-Fluorouracil (5FUra) is the third most popular chemotherapeutic component employed to treat solid tumors. In the present study, we aimed to appraise the silymarin (SM) and silymarin nanoemulsion (SMN) effect on 5FUra-induced gastrointestinal toxicity in adult male rats. A total of 30 male Wistar rats were divided into 6 groups including the control (Crl) group, and groups treated with SMN (5 mg.kg−1), SM (5 mg.kg−1), 5FUra + SMN (5 mg.kg−1), and 5FUra + SM (5 mg.kg−1) by IP injection for 14 days. And gastrointestinal toxicity was induced by a single intraperitoneal (IP) injection of 5FUra (100 mg.kg−1) for the last group in the study. Treating rats with SM and SMN diminished elevating malondialdehyde (MDA) levels, and improved total antioxidant capacity (TAC) levels. Also, the intensity of mRNA expression of interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-α) caused by 5FUra in the gastrointestinal tissue tract, and macroscopic oral ulcerations decreased, ass well as weight loss was prevented, particularly in the SMN group. Moreover, in the microscopic scope, there were significant improvements in the levels of hyperemia, hyaline, and inflammatory cell infiltration in the tongue, esophagus, and intestinal tissues in the FUra + SMN and FUra + SM groups compared to 5FUra. Hence, treatment with SM and SMN reduced oxidative stress, histopathological degeneration, and gene expression of inflammatory markers in the gastrointestinal tract. According to the results, treatment with SM and SMN markedly decreases the gastrointestinal toxicity caused by 5FUra.

Association of Undiagnosed Pre-Diabetes and Type-2 Diabetes Mellitus with Interleukin-2 mRNA Expression Among Adults in Bayelsa State, Nigeria

Many cases of hyperglycemia are undiagnosed in Nigeria due to poverty and inadequate access to health care. Although hyperglycemia and type 2 diabetes mellitus (T2DM) are inflammatory conditions, however, the role of inflammatory mediators such as interleukin-2 in hyperglycemia and T2DM is not fully understood. The aim of the present study was to evaluate the prevalence and association of undiagnosed pre-diabetes and T2DM with IL-2 mRNA level among adults in Bayelsa State, Nigeria. A cross sectional design was employed to randomly select 130 adults and they were grouped into three: normal control, prediabetes, and T2DM. Fasting blood glucose concentration was measured using Accu-answer glucometer, while mRNA level of IL-2 was quantified using real time PCR method. The prevalence of undiagnosed pre-diabetes was 43.07% (males, 19.23%; females, 23.85%) while undiagnosed T2DM was 16.15% (males, 6.92%; females 9.23%). The expression of IL-2 mRNA was significantly upregulated in both pre-diabetes and T2DM subjects compared with normal control (p &lt; 0.05), but was significantly higher among T2DM subjects. Age ≥ 50 years, physical inactivity and marriage was significantly associated with hyperglycemia and T2DM (p &lt; 0.05). The present study found a high prevalence of undiagnosed pre-diabetes and T2DM among adults in Bayelsa State, Nigeria. Association of prediabetes and T2DM with IL-2 mRNA expression was found. IL-2 mRNA level might be a useful biomarker for monitoring diabetes mellitus progression. Directing diabetes survey and interventions to rural communities is highly recommended. Keywords: [Undiagnosed Diabetes, pre-diabetes, IL-2, rural communities, Bayelsa State, Nigeria]

Expression analysis of IL-2, TBX21 and SOCS1 in peripheral blood cells of celiac disease patients reveals the diagnostic potential of IL-2.

Celiac disease (CD) is a chronic immune-mediated enteropathy and a cytokine network is involved in its pathogenesis. Interleukin-2 (IL-2) has a key role in the adaptive immune pathogenesis of CD and has been reported to be one of the earliest cytokines to be elicited after gluten exposure by CD patients. This study aimed at investigating the expression level of IL-2 and functionally related genes SOCS1 and TBX21 in active and treated CD patients compared to controls. Peripheral blood (PB) samples were collected from 40 active CD (ACD), 100 treated CD, and 100 healthy subjects. RNA was extracted, cDNA was synthesized and mRNA expression levels of the desired genes were investigated by Real-time PCR. The gene-gene interaction network was also constructed by GeneMANIA. Our results showed a higher PB mRNA expression of IL-2 in ACD patients compared to controls (p = 0.001) and treated CD patients (p˂0.0001). The mRNA expression level of TBX21 was also significantly up-regulated in ACD patients compared to controls (P = 0.03). SOCS1 mRNA level did not differ between active and treated CD patients and controls (p˃0.05) but showed a significant correlation with the patient's aphthous stomatitis symptom (r = 0.37, p = 0.01). ROC curve analysis suggested that the use of IL-2 levels can reach a high specificity and sensitivity in discriminating active CD patients. The PB level of IL-2 has the potential to be introduced as a diagnostic biomarker for CD. Larger cohort studies, including pediatric patients, are needed to achieve more insights in this regard.

Berberine promotes M2 macrophage polarisation through the IL-4-STAT6 signalling pathway in ulcerative colitis treatment

AimThis study focusses on the anti-inflammatory and immune-modulatory roles of berberine (BBR) in ulcerative colitis (UC) treatment. Additionally, the underlying mechanisms of BBR were systematically explored. MethodsA 3% (w/v) dextran sodium sulphate (DSS) solution was used for establishing the mice UC model. M2 macrophage polarisation was induced in RAW 264.7 cells using interleukin 4 (IL-4), whereas M1 macrophage polarisation was induced using lipopolysaccharide. Colon length, colon mucosa damage index (CMDI), and haematoxylin–eosin (HE) staining were used to evaluate colon damage induced by DSS. M1/M2 macrophages in the colon tissue were identified using immunofluorescence (IF) staining with CD86+ or CD163+. M1/M2 macrophages in the abdomen were examined using flow cytometry. An enzyme-linked immunosorbent assay was conducted to identify M1/M2 macrophage supernatant biomarkers in RAW 264.7 cells. Western blotting, immunohistochemical staining, and real-time PCR were performed to investigate the potential mechanisms of BBR for treating UC in vivo and in vitro. ResultsBBR was found to prolong colon length, ameliorate CMDI and alleviate the colon's pathological changes in UC mice. In DSS-induced UC mice, M1 macrophages predominated. BBR promoted M2 macrophages and suppressed M1 macrophages in the colon and abdomen of DSS-induced UC mice. Additionally, BBR significantly decreased M1-specific markers (IFN-γ and IL-1β) while increasing M2-specific markers (IL-10 and TGF-β) in the supernatants of RAW 264.7 cells. BBR upregulated the mRNA expression of IL-4, STAT6, and Chil3 while downregulating TNF-α, IFN-γ, and NOS2 expression in vivo. Moreover, BBR activated the downstream targets of the IL-4-STAT6 signalling pathway and enhanced the phosphorylation of STAT6 in vivo and in vitro to polarise M2 macrophage. ConclusionIn UC mice, BBR suppressed M1 macrophages while promoting M2 macrophages. M1 macrophage suppression and M2 macrophage activation were strongly correlated with the anti-inflammatory and immune-modulating activities of BBR. BBR induced the polarisation of M2 macrophages by activating the IL-4-STAT6 signalling pathway, which contributed to its therapeutic efficacy against UC.

Multi-component immune knockout: A strategy for studying the effective components of traditional Chinese medicine

Periploca forrestii Schltr., a traditional Chinese medicine (TCM), is commonly used to treat autoimmune diseases such as rheumatoid arthritis (RA). However, its mechanism, involving a variety of cardiac glycosides, remains largely unknown. The immune knockout strategy can highly selectively deplete target components by immunoaffinity chromatography (IAC). We aimed to identify the common structural features of cardiac glycosides in P. forrestii and design IAC to specifically recognize these features to achieve the multi-component knockout of potential active substances from the extracts of P. forrestii. A content detection experiment confirmed that the content of a compound with periplogenin structure (CPS) in the extract of P. forrestii was reduced by 45% by IAC of periplogenin. The immunosuppressive ability of the extract on H9 human T lymphocytic cells was weakened after CPS knockout from P. forrestii extract. Molecular biology experiments showed that mRNA expression of interferon-γ (IFN-γ), interleukin-2 (IL-2), and interleukin-6 (IL-6) in H9 cells was up-regulated after CPS knockout, while no significant changes in the expression of interleukin-4 (IL-4) were found. CPS knockout from P. forrestii extract did not cause significant changes in the proliferation of lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells incubated with this extract. These results indicate that CPS exhibited immunosuppressive effects via inhibiting the T helper 1 (Th1) cell immune response and not via the anti-inflammatory components in P. forrestii. This is the first use of IAC to achieve multi-component knockout in TCM extracts for identifying effective compounds. This method is effective and reliable and warrants further exploration.

Cold Drinking Water Boosts the Cellular and Humoral Immunity in Heat-Exposed Laying Hens

This study aimed to investigate the effects of cold drinking water on cellular and humoral immunity in heat-exposed laying hens. One hundred and eight laying hens at 19 weeks old were placed into three treatments with six replicates of six hens in each group as follows: (1) hens were provided with normal drinking water (NW) under the control of thermoneutral temperature (CT: 25 ± 1 °C; CT + NW), (2) hens were provided with NW under high ambient temperature (HT: 35 ± 1 °C; HT + NW) for 8 h/d for a month, and (3) hens were treated under HT with cold drinking water (CW: 15 ± 1 °C; HT + CW) for 8 h/d for a 4-weeks. Then, the feed consumption, egg production, egg weight, feed conversion ratio, and blood immune parameters were investigated. The results showed that cold drinking water (CW) caused a significant (p < 0.05) recovery in the reduction of food intake and egg production due to heat stress; however, there was no significant effect (p > 0.05) on egg weight and feed conversion ratio. Moreover, CW significantly (p < 0.05) restored the immune-suppressing effects of heat stress on the contents of peripheral blood mononuclear cells, including B-cell (BU-Ia), helper T cell (CD4), and the ratio of helper/cytotoxic T cell (CD4/CD8). In addition, CW significantly (p < 0.05) recovered the reduction on the level of mRNA expression of interleukin-2 (IL-2) and interferon-gamma (IFN-γ), as well as significantly (p < 0.05) restored the reduction of plasma concentration of IL-2, IFN-γ and immunoglobulin G in heat-stressed laying hens. These results prove that CW increased heat dissipation and enhanced feed intake, egg production, and cellular and humoral immunity in heat-exposed laying hens.

Anti-inflammatory effects of the prostacyclin analogue iloprost in an in vitro model of inflamed human dental pulp cells.

Iloprost's anti-inflammatory effects on human dental pulp stem cells (HDPCs) are currently unknown. We hypothesized that iloprost could downregulate the expression of inflammatory-related genes and protein in an inflamed HDPC in vitro model. To induce inflammation, the HDPCs were treated with a cocktail of interleukin-1 beta, interferon-gamma, and tumour necrosis alpha, at a ratio of 1:10:100. Iloprost (10-6 M) was then added or not to the cultures. Interleukin-6 (IL-6) and interleukin-12 (IL-12) mRNA expression were assessed by real-time polymerase chain reaction. IL-6 protein expression was assessed by enzyme-linked immunosorbent assay. The results were analysed using one-way ANOVA or the Kruskal-Wallis test. The cytokine cocktail induced more robust IL-6 expression than LPS treatment. Iloprost slightly, yet significantly, downregulated IL-6 and IL-12 mRNA expression. These findings suggest that iloprost might be used as a beneficial component in vital pulp therapy.

Immunomodulatory Effects of Chicken Broth and Histidine Dipeptides on the Cyclophosphamide-Induced Immunosuppression Mouse Model.

The carnosine and anserine, which represent histidine dipeptides (HD), are abundant in chicken broth (CB). HD are endogenous dipeptide that has excellent antioxidant and immunomodulatory effects. The immunomodulatory effect of CB hydrolysate (CBH) and HD in cyclophosphamide (CTX)-induced immunosuppressed mice was examined in this study. CBH and HD were given to mice via oral gavage for 15 days, accompanied by intraperitoneal CTX administration to induce immunosuppression. CBH and HD treatment were observed to reduce immune organ atrophy (p &lt; 0.05) and stimulate the proliferation of splenic lymphocytes (p &lt; 0.05) while improving white blood cell, immunoglobulin M (IgM), IgG, and IgA levels (p &lt; 0.05). Moreover, CBH and HD strongly stimulated interleukin-2 (IL-2) and interferon-gamma (IFN-γ) production by up-regulating IL-2 and IFN-γ mRNA expression (p &lt; 0.05) while inhibiting interleukin-10 (IL-10) overproduction and IL-10 mRNA expression (p &lt; 0.05). In addition, CBH and HD prevented the inhibition of the nitric oxide (NP)/cyclic guanosine monophosphate-cyclic adenosine monophosphate (cGMP-cAMP)/protein kinase A (PKA) signaling pathway (p &lt; 0.05). These results indicate that CBH and HD have the potential to prevent immunosuppression induced by CTX. Our data demonstrate that CBH can effectively improve the immune capacity of immunosuppressed mice similar to the same amount of purified HD, which indicates that CBH plays its role through its own HD.

Effectiveness of Indonesian house dust mite allergenic extract in triggering allergic rhinitis sensitivity in a mouse model: A preliminary study

Perennial allergic rhinitis (AR) is a chronic upper respiratory disease, with inflammation mediated by immunoglobulin E in the nasal mucosa caused by house dust mites. Recently, allergen immunotherapy showed promising allergic healing in patients with a definite history of sensitization. Based on this finding, a product was developed using Indonesian house dust mite (IHDM). This study aimed to optimize the allergenic rhinitis mouse model that was generated using IHDM to test the in vivo sensitivity and safety of this product. Seven groups of mice were used for effectiveness testing - normal, negative control with IHDM challenge, positive control with 0.1% histamine challenge, and AR group by both IHDM-induced sensitization at 12.5, 50, 250, or 500 μg and IHDM challenge. Mice were sensitized by intraperitoneal administration of IHDM once a week for 3 consecutive weeks. Thereafter, the challenge was given intranasally 5 times on alternate days. The number of nose rubbing and sneezing was noted. Eosinophil infiltration was assessed histologically using hematoxylin and eosin staining. The expression of interleukin-5 (IL-5) mRNA in the nasal mucosa was determined using semi-quantitative reverse transcription-polymerase chain reaction. The induction of AR with IHDM significantly increased the number of nose rubbing and sneezing in the mouse model. Eosinophil infiltration was observed in the nasal mucosa; however, no significant change occurred in the expression of IL-5 mRNA. Overall, these data indicate that IHDM allergenic extract could be an effective sensitizing agent in a mouse model of AR. Although the use of IHDM is a limitation of this study because other sources of house dust mites might have different effects, this study provides a proper model for immunotherapy effectivity testing for in vivo pre-clinical studies.

Effects of different levels of rutin on growth performance, immunity, intestinal barrier and antioxidant capacity of broilers

This study investigated the effects of different doses of rutin on the growth performance, immunity, intestinal barrier, and antioxidant capacity of broilers. A total of two hundred and fifty-six 1-day-old male broilers were divided into 4 groups and fed basal diets supplemented with 0 (control group), 250, 500 and 1,000 mg rutin/kg, respectively, for 42 days. In the starter period (days 1–21) and whole trial period (days 1–42), broilers fed diets containing 500 mg rutin/kg had significantly (p < 0.05) higher average daily feed intake, body weight and average daily gain and better feed conversion ratio. Dietary 500 mg rutin/kg increased the villus height, villus height to crypt depth ratio and villus area, while reducing the B cell lymphoma 2 associated X mRNA expression (p < 0.05). Dietary 500 mg rutin/kg increased the level of IgA in serum and decreased serum tumour necrosis factor-α (TNF-α) content, and the nuclear factor kappa-B, interleukin-2 and TNF-α mRNA expression in jejunal mucosa (p < 0.05). Simultaneously, 500 and 1,000 mg rutin/kg significantly decreased serum diamine oxidase activity, D-lactic acid and lipopolysaccharide concentration, increased activities of total superoxide dismutase and total antioxidant capacity in jejunal mucosa (p < 0.05), and upregulated mRNA expressions of genes related to intestinal barrier and antioxidant capacity in jejunal mucosa (p < 0.05). These findings indicate that dietary rutin, especially at 500 mg/kg, improves broilers’ growth performance, intestinal barrier function, immunity, and antioxidant capability, which may be related to the nuclear factor erythroid-2-related factor 2 (Nrf2) pathway. Highlights Dietary rutin improved growth performance, jejunal morphology, and intestinal barrier function. Dietary rutin enhanced the immunity via inhibiting NF-κB, and improved antioxidant capacity via activating Nrf2/HO-1 pathway in broilers. The optimal dose of rutin was 500 mg/kg in broiler diet.

Eosinophilic Esophagitis: Cytokines Expression and Fibrotic Markers in Comparison to Celiac Disease.

Introduction: Eosinophilic esophagitis (EoE) is now recognized as the main inflammatory condition that leads to fibrosis, unlike other chronic inflammatory gastrointestinal diseases, such as celiac disease. The aim of our study is to characterize the collagen deposition and cytokine expression involved in the fibrogenic response in patients affected by EoE in comparison to celiac disease. Materials and Methods: Consecutive patients with a clinical suspicion of untreated EoE or active celiac disease were enrolled. In the control group, patients with negative upper endoscopy were included. Total RNA was isolated from biopsy specimens using a commercial kit (SV Total RNA Isolation System, Promega Italia Srl). Quantitative real-time PCR (qRT-PCR) was performed in triplicate using a StepOne™ Real-Time PCR instrument (Thermo Fisher Scientific, Monza, Italy). mRNA encoding for inflammatory molecules: interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 13 (IL-13), and fibrotic markers: transforming growth factor beta 1 (TGF-β), mitogen-activated protein kinase kinase kinase 7 (MAP3K7), serpin family E member 1 (SERPINE1), were quantified using TaqMan Gene Expression Assays (Applied Biosystems). RESULTS. In EoE, the qPCR analysis showed an increase in all the inflammatory cytokines. Both IL-5 and Il-3 mRNA expression resulted in a statistically significant increase in oesophageal mucosa with respect to the celiac duodenum, while no differences were present in IL-4 expression. TGF-β expression was similar to the controls in the mid esophagus but reduced in the distal EoE esophagus (RQ: 0.46 ± 0.1). MAP3K7 expression was reduced in the mid esophagus (RQ: 0.59 ± 0.3) and increased in the distal esophagus (RQ: 1.75 ± 0.6). In turn, the expression of SERPINE1 was increased in both segments and was higher in the mid than in the distal esophagus (RQ: 5.25 ± 3.9, 1.92 ± 0.9, respectively). Collagen deposition was greater in the distal esophagus compared to the mid esophagus [18.1% ± 8 vs. 1.3% ± 1; p = 0.008]. Conclusions: The present study confirms the esophageal fibrotic involution involving the distal esophagus and shows that the inflammatory pathway in EoE is peculiar to this disease and different from other chronic inflammatory gastrointestinal disorders such as celiac disease.

Porcine reproductive and respiratory syndrome virus reinfection causes the distribution of porcine interleukin-4 in close proximity to B lymphocytes within lymphoid follicles and a reduction in B and T lymphocytes

Interleukin 4 (IL-4) plays a major role in T-lymphocyte development and is thought to be a central regulator as a cofactor in resting B-lymphocyte proliferation. Primary infection with porcine reproductive and respiratory syndrome virus (PRRSV) induces minimal IL-4 production, whereas an IL-4 response occurs in the peripheral blood of piglets reinfected by PRRSV. The locations and interaction partners for the massive volume of IL-4 triggered by PRRSV reinfection remain unclear. This study aimed to investigate the characteristics of IL-4 secretion and location changes in peripheral immune organs induced by PRRSV infection and reinfection. Our results show that PRRSV reinfection induced higher levels of IL-4 mRNA and protein expression in the peripheral immune organs (e.g., lymph node and spleen) and peripheral blood compared with PRRSV primary infection. Importantly, we found that, following PRRSV reinfection, an obvious large-scale migration of IL-4 occurred in the lymph nodes. During PRRSV primary infection, IL-4 was mainly concentrated around the lymphoid follicles and paracortical regions of the lymph node and also located in the marginal area and periarterial lymphatic sheath region of the spleen. During PRRSV reinfection, the now abundant IL-4 gathered into the lymphoid follicles of the lymph node and spleen. Notably, IL-4 changed its location state from scattered and sparse during primary infection to clinging to B lymphocytes in the lymphoid follicles during reinfection. During reinfection, IL-4 was often co-localized with T and B lymphocytes; furthermore, the percentages of several T lymphocyte subsets, N protein-specific antibody levels, and viral load in the peripheral blood or lymph tissues underwent remarkable variation. Another important finding of this study was that the numbers of B lymphocytes and T lymphocytes in the lymphoid nodes were significantly reduced after PRRSV infection or reinfection, presumably due to PRRSV-induced acute bone marrow failure and autophagy in thymic epithelial cells. This study revealed the characteristics of IL-4 migration and distribution in the peripheral lymph organs induced by PRRSV reinfection and provides valuable clues for further exploration of the interactions between IL-4, B lymphocytes, and T lymphocytes during PRRSV infection and reinfection.

Immunomodulatory Effect of Ginsenoside Rb2 Against Cyclophosphamide-Induced Immunosuppression in Mice.

Ginsenoside Rb2 (Rb2), a fundamental saponin produced and isolated from ginseng (Panax ginseng C.A. Meyer), has a wide range of biological actions. The objective of this investigation was to see if ginsenoside Rb2 has any immunomodulatory properties against cyclophosphamide (CTX)-induced immunosuppression. For the positive control group, levamisole hydrochloride (LD) was used. We discovered that intraperitoneal injection of Rb2 (5, 10, 20 mg/kg) could relieve CTX-induced immunosuppression by enhanced immune organ index, reduced the pathological characteristics of immunosuppression, promoted natural killer (NK) cells viability, improved cell-mediated immune response, boosted the IFN-γ (Interferon-gamma), TNF-α (Tumor necrosis factor-alpha), IL-2 (Interleukin-2), and IgG (Immunoglobulin G), as well as macrophage activity like carbon clearance and phagocytic index. Rb2 significantly elevated the mRNA expression of IL-4 (Interleukin-4), SYK (Tyrosine-protein kinase-SYK), IL-2, TNF-α, and IL-6 (Interleukin-6) in the spleen of CTX-injected animals. Molecular docking results showed that Rb2 had excellent binding properties with IL-4, SYK, IL-2, TNF, and IL-6, indicating the target protein might be strongly correlated with the immunomodulatory effect of Rb2. Taken together, ginsenoside Rb2 can improve the immune function that is declined in CTX-induced immunosuppressed mice, the efficacy maybe due to the regulation of related cytokine and mRNA expression.

Acupoint catgut embedment may reduce airway inflammation reaction by down-regulating ICAM-1 and EOS by suppressing p38MAPK signaling in lung tissue of asthmatic rats

To observe the effect of acupoint catgut embedment(ACE) on expression of p38 mitogen activated protein kinase (p38MAPK), intercellular adhesion molecule-1(ICAM-1), interleukin-4 (IL-4) and eosinophils (EOS) in lung tissue of asthmatic rats, so as to explore its mechanism underlying improvement of asthma. Forty male Wistar rats were randomly divided into control, model, ACE and dexamethasone (DEX) groups, with 10 rats in each group. The asthmatic model was established by intraperitoneal injection of mixture suspension (1 mL) of ovalbumin (OVA,10%) and 10% Al (OH)3+ normal saline, followed by inhalation of atomized 1% OVA solution for 30 min, once daily for 2 weeks to trigger occurrence of asthmatic symptoms. The ACE was applied once to "Feishu" (BL13), "Dingchuan" (EX-B1) and "Danzhong" (CV17). Rats of the DEX group were given intraperitoneal injection of DEX once a day for 2 weeks. H.E. staining was used to evaluate histopathological changes of the lung tissue. The relative number of EOS in the bronchoalveolar lavage fluid (BALF) was detected by Wright Giemsa staining. The apoptosis level of EOS in the lung tissue was detected by TUNEL staining. The ultrastructural changes of EOS in the lung tissues were observed by transmission electron microscope (TEM). The expression of p38MAPK, ICAM-1 and IL-4 mRNAs in the lung tissue was detected by quantitative real-time PCR. Findings of optical microscope and TEM showed obvious bronchial deformation and inflammatory cell infiltration, rupture of EOS cell membrane, uneven cytoplasm with swelling and uneven density of eosinophilic granules in EOS of the model group, which was relatively milder in the ACE and DEX groups. Compared with the control group, the EOS number in BALF and the expressions of p38MAPK, ICAM-1 and IL-4 mRNAs in the lung tissue were significantly increased (P<0.01), and the apoptosis index of EOS in the lung tissue was significantly decreased (P<0.01) in the model group. After intervention, the EOS number in BALF and expression levels of p38MAPK, ICAM-1 and IL-4 mRNAs in the lung tissue of ACE and DEX groups were significantly decreased (P<0.01, P<0.05), as well as the apoptosis index of EOS in the lung tissue was significantly increased in both ACE and DEX groups (P<0.01) in comparison with the model group. The EOS number in BALF and expression of ICAM-1 mRNA were significantly lower in the DEX group than those in the ACE group (P<0.05). Catgut embedding at acupoints may alleviate the airway inflammatory response in asthma rats, which may be related with its effects in down-regulating p38MAPK signaling, ICAM-1 and IL-4 mRNA expression, reducing the aggregation of EOS, and promoting the apoptosis of EOS.