Lymphocyte Proliferation Activity Research Articles

Stability of oil-in-water emulsion and immunomodulating activity in S180 tumor-bearing mice.

The stability and nutritional integrity of emulsions are susceptible to various factors including thermal treatment, solid-liquid ratio, and sterilization. In this study, the physicochemical stability and immunomodulatory activities of an oil-in-water emulsion containing immune peptides (TUFSE) were assessed through particle size, zeta potential, related cytokines, and so on. When the temperature was 70°C and a solid-liquid ratio of 1:4, the emulsion revealed stability at high-pressure homogenization, with the small particle size. The loss rates of vitamins were 8.57%-62.26% in 6 months at 25°C. After treatment with cyclophosphamide (CTX), lymphocyte proliferation activity in TUFSE-H group increased (p<0.05), and immune globulin levels were notably elevated (p<0.05) in TUFSE groups compared to model group. It confirms the parameters of the emulsion, suggesting its ability to be prepared with minimal vitamin loss while simultaneously improving the disease status in CTX-treated tumor-bearing mice. It shows potential as an immune-enhancing supplement with significant potential value. PRACTICAL APPLICATION: This study validated the parameters of the oil-in-water emulsion and showed that it can be stably prepared with minor vitamin loss while simultaneously improving the disease status in CTX-treated tumor-bearing mice. TUFSE-H group exhibited a notable increase in lymphocytes proliferation activity, whereas serum cytokines and immune globulin levels were elevated compared to MC group, indicating its potential as an immune-enhancing supplement with substantial value.

Preparation Optimization and Immunological Activity Studies of Portulaca oleracea L. Polysaccharides Liposomes.

This study combines traditional Chinese medicine polysaccharides with nanomaterials to enhance drug bioavailability and immunological activity. The study of polysaccharide preparation, structure identification, pharmacological activity, and mechanism of action is deepening, but the research combined with the new drug delivery system is relatively weak, so the application of polysaccharides is still facing great limitations. In order to prolong the action time of polysaccharides and improve their bioavailability, liposome has become the most promising delivery carrier. The purpose of this study was to optimize the preparation process of Portulaca oleracea L. polysaccharides liposomes (POL-PL) and evaluate the immunoactivity in vitro. POL-PL was prepared by reverse evaporation, and the preparation process was optimized using the response surface methodology. The characteristic analysis of POL-PL was detected by the indicators including morphology, particle size, zeta potential, encapsulation efficiency, release, and stability. The effects of POL-PL on the proliferation and immunological activity of mouse spleen lymphocytes and RAW264.7 cells were evaluated in vitro. POL-PL is highly homogeneous in morphology and particle size, and its sustained release improves the bioavailability of Portulaca oleracea L. polysaccharides (POL-P). Moreover, POL-PL treatment significantly enhanced the proliferation and phagocytic activity of RAW264.7 cells and increased the secretion of IL-6, TNF-α, IL-1β, and NO. This study suggested that POL-PL were prepared successfully by reverse evaporation method, and POL-PL had immunoenhancing activity in vitro. The results provided a theoretical basis for further application of POL-PL.

Hepatic P53 upregulation and the genotoxic potential of acesulfame-K treatment in rats with a special emphasis on in vitro lymphocyte and macrophage activity testing.

Acesulfame-k (Ace-k) is a widely used artificial sweetener in various products, and long-term cumulative and multisource exposure is possible despite inadequate toxicological data confirming its safety. Ninety male rats were divided into two main groups according to their body weight into immature and mature rats. Each group was subdivided into 3 subgroups: control untreated, 30 and 90mg/kg b. w of Ace-k via gastric intubation. The treatment was performed daily 5days per week for 12weeks. At the end of the experimental period, blood samples were collected for in vitro testing of lymphocyte proliferation rate, comet assay, and macrophage activity about nitric oxide (NO) production. In addition, the collection of liver specimens was performed for P53 gene expression and histopathological evaluation. The results revealed that Ace-k induced modulation in lymphocyte proliferation rate and affected the production of NO by macrophages while increasing in tail moment in a dose-dependent manner that varied among different age groups. The upregulation of P53 in the liver was correlated with increased polyploidization and necro apoptotic reaction and various histopathological hepatic alterations. The present data revealed that chronic treatment of rats with Ace-k affects lymphocyte proliferation and macrophage activity in a dose-dependent manner. In addition, the genotoxic and hepatotoxic potential of Ace-k were confirmed.

Влияние никотина на систему клеточного иммунитета у мужчин с хронической обструктивной болезнью легких

INTRODUCTION: One of common pulmonary diseases associated with tobacco smoking, is chronic obstructive pulmonary disease (COPD). Despite a large number of publications devoted to the problem of COPD, there remain many unclear issues concerning the involvement of the immune system in the development of the disease. The effect of nicotine on cell-mediated immunity in COPD has not been studied in practice. AIM: To study the influence of nicotine on the proliferating activity of lymphocytes of the peripheral blood in response to phytohemagglutinin (PHA) in patients with COPD. MATERIALS AND METHODS: Fifty two patients with COPD were examined. The control group included 32 patients of similar gender and age, without any significant history of smoking and of respiratory diseases. All the patients completed CAT questionnaire and were tested on mMRC scale, and underwent spirometry, examination for the proliferating activity of lymphocytes in response to phytohemagglutinin and for the functional activity of cells synthesizing cyclooxygenase. RESULTS: In patients with COPD, a decrease in the proliferative activity of lymphocytes in response to PHA was found in comparison with this parameter of patients without COPD (50.4 ± 10.2% vs. 55.0 ± 10.2%, p &lt; 0.05), as well as an increase in the activity of cells synthesizing cyclooxygenase (1.2 ± 0.3 vs. 1.0 ± 0.2 p &lt; 0.001). Addition of a combination of nicotine and PHA to cell culture led to a decrease in the proliferative activity of cells in response to mitogen in most patients, and to its increase in a third of patients. CONCLUSION: In patients with COPD with the underlying inflammation, suppression of cell-mediated immune reactions was noted. Nicotine in vitro suppresses the response of lymphocytes to mitogen in most patients, however, in some patients it causes stimulation, the cause of which remains unclear.

The signal pathway of melatonin mediates the monochromatic light-induced T-lymphocyte apoptosis in chicken thymus

Our previous study revealed that under monochromatic red light (RL), the melatonin nuclear receptor reduces the proliferation activity of broiler thymic lymphocytes through the P65 signaling pathway. The main objective of this study was to investigate the signal mechanism by which RL decreases thymic lymphocyte proliferation. Initially, broilers were purchased and randomly assigned to be fed under white light (WL), red light (RL), green light (GL), and blue light (BL). Pinealectomy was performed 3 d later, and the broilers were euthanized after 14 d. The results showed that the expression of the antiapoptotic proteins Bcl-2/Bcl-xl decreased under RL, while the expression of the pro-apoptotic factor Bax/caspase-3 and the pro-inflammatory factors INF-γ/TNF-α/IL-6 increased. After pinealectomy, the expression of Bax/TNF-α/IL-6 increased in conjunction with the decrease in Bcl-2 expression. In vitro experiments demonstrated that exogenous melatonin decreased the expression of Bax/TNF-α/IL-6 in thymic lymphocytes of chicks reared under RL. This melatonin-induced effect was enhanced by SR1078 (RORα/RORγ agonist) but attenuated by SR3335 (RORα antagonist) and BAY (P65 antagonist). These findings revealed that the melatonin nuclear receptor RORα/RORγ promotes the expression of the pro-apoptotic factor Bax/caspase-3 and the pro-inflammatory factors INF-γ/TNF-α/IL-6, while inhibiting the expression of the antiapoptotic factor Bcl-2/Bcl-xl. Our research suggested the signaling pathway of monochromatic red light impacts the apoptosis of thymus lymphocytes in broiler.

Dietary propolis improves the growth performance, redox status, and immune response of Nile tilapia upon a cold-stress challenge.

The purpose of this research was to demonstrate the potential of adding propolis (PR) to the diet of Nile tilapia (Oreochromis niloticus) to mitigate the harmful effect of cold stress (CS) on the growth performance, redox status, and immunological response. Two trials were conducted in this study. First, 210 Nile tilapia fingerlings (28.61±0.20 g) were used in a preliminary trial to determine the appropriate PR level and supplementation period to be applied for the main trial. Fish were assigned into 7 treatment groups (3 aquaria replicates × 10 fish per aquarium in each treatment group) according to the rate of PR supplementation in the fish diets at 0, 2, 4, 6, 8, 10, and 12 g/kg for 6 consecutive weeks. The average body weight and body weight gain were determined weekly. It was found that PR supplementation at 10 g/kg in fish diet for 4 weeks was enough to obtain significant results on the growth performance of Nile tilapia. For the main trial of the present study, 480 Nile tilapia fingerlings (average weight 29.93±0.11 g) were distributed into randomized 2 PR × 2 CS factorial treatment groups (6 replicate aquariums containing 20 fish in each group). Fish of PR groups received a basal diet for a feeding period of 4 weeks, included with 10 g/kg PR (+ PR group) or without PR inclusion (- PR group). Fish of the CS groups were either challenged with cold stress at 18°C (+ CS group) or maintained at a temperature of 26°C during the feeding period (- CS group). The results showed that CS challenge significantly (p < 0.05) impaired the growth indices, redox status, and immune response in the challenged fish compared to the non-challenged fish. On contradictory, the inclusion of PR into fish diets enhanced (p < 0.05) the feed intake, growth indices, antioxidant enzyme activity, and immunological parameters. Moreover, PR treatment alleviated the CS deterioration of fish weights, specific growth rates, feed efficiency, antioxidant enzyme activity, lymphocyte proliferation, and phagocytosis activity and alleviated the elevated mortality, H/L ratio, and malondialdehyde levels by cold stress. It is concluded that the inclusion of propolis at 10 g/kg in the diet of Nile tilapia fish could be approved as a nutritional approach to enhance their performance, especially when stressed by low-temperature conditions.

The effect of COVID-19 infection on the white blood cell count and lymphocyte proliferation activity at the early stage of the disease

Since the appearance of COVID-19 at the end of December 2019 in Wuhan, China, and its prevalence in many countries, the symptoms of this disease extended from respiratory problems to a wide range of symptoms associated with the invasion of the virus to many organs and tissues in the body of patients. The white blood cells, particularly T lymphocytes, are the main effectors in defense against viral infections. This study was performed to investigate the response of white blood cells to the infection of SARS-COV-2 at the early stage of the disease. T cells decreased in number in the circulation, but this decrease was not associated with an impairment of their activity. Moreover, stimulation of virus-infected T cells with nonspecific mitogen revealed increased cell proliferation. This study concluded that T lymphocytes are highly activated during SARS-COV-2 infection, despite lymphopenia, at least at the early stages of the disease. Keywords: COVID-19, lymphocyte proliferation, mitotic index

Combination of Probiotics and Resveratrol to Target Drug Resistance in Breast Cancer: An In vitro and In vivo Study

Background: Probiotics exhibit significant activity in the immune system by activating multiple immune mechanisms. Resveratrol is a non-flavonoid polyphenol. It has various pharmacological effects, including anti-oxidative, anti-inflammatory, anti-proliferative, and anti-angiogenesis effects. Objectives: The present study investigates the potential immunomodulatory and antitumor activity of the combination treatment of probiotics and resveratrol in sensitive and cisplatin-resistant breast cancer. Methods: In the in vivo study, tumor-bearing mice received one of the following treatments: 2.5*108 CFU/ml probiotics, 50 mg/kg resveratrol, the combination of probiotics and resveratrol, vehicle, or cisplatin. Balb/C mice were inoculated with sensitive EMT6/P and cisplatin resistance EMT6/CPR cancer cell lines, and in vivo antitumor activity was evaluated. Results: The antiproliferative activity of the probiotics, resveratrol, and their combination treatments was assessed using an MTT assay to evaluate lymphocyte proliferation activity. LDH colorimetric assay was conducted to measure the effectiveness of the treatments on the activity of natural killer cells. Nitro blue tetrazolium assay and neutral red method were used to evaluate macrophage function. Conclusion: The combination treatment showed an enhanced effect in splenic lymphocyte proliferation, macrophage function, phagocytosis, and pinocytosis in both cell lines. A significant reduction in tumor size and weight in EMT6/P and EMT6/CPR-bearing mice occurred. Hence, the combination treatment of probiotics and resveratrol was found to have a valuable activity against sensitive and cisplatin-resistant breast cancer cells and might act as a stimulator of the immune system. Therefore, the combination of probiotics and resveratrol deserves further analysis to be used in cancer prevention and treatment.

Immunomodulatory and neurotropic activities of synthetic peptides in a model of brain injury in rats

Treatment of consequences of traumatic brain injury (TBI) remains one of the current problems of medicine. To increase the effectiveness of treatment of post-traumatic complications, various drugs are recommended, including the peptide with neuromodulatory activity Semax.The present study aims to determine the presence of neuro- and immunoprotective properties of the synthetic peptide PR5, composed of fragments of proline-rich antimicrobial peptides.The work was performed on male Wistar rats weighing 300-350 g. The “falling weight” model of mechanical brain injury was used, which mainly causes diffuse brain damage. The synthesized peptide PR5, composed of fragments of known proline-rich peptides of animal neutrophils, and the peptide preparation Semax in the form of a 1% aqueous solution were used. The drugs were administered intranasally 1 hour after TBI, then twice a day for 4 days at a dose of 100 mg/kg body weight. Control animals received physiological saline in the same regimen as the peptide preparations.TBI led to a significant decrease in body weight, but in rats receiving the peptide preparation Semax, the decrease in body weight was significantly less than in control animals, and the PR5 preparation completely prevented the decrease in body weight after TBI. After TBI, the proliferative activity of lymphocytes was suppressed and the cytotoxicity of NK cells decreased. In animals treated with peptide preparations, there was no significant suppression of NK cell cytotoxicity, and the proliferative activity of lymphocytes was restored to the level of control animals by day 14 after TBI. Both peptide preparations used contributed to higher locomotor activity, and in animals treated with the PR5 peptide, this type of activity reached the parameters of control animals. The reduction in freezing duration in groups treated with peptide preparations indicates the presence of a sedative effect.The peptide preparation PR5 was active in this series of experiments, showing immunotropic and neuroprotective activity comparable to the Semax preparation. Further studies aimed at confirming the identified types of activity of the peptide preparation PR5 may justify its prospects for clinical use as a new nootropic agent.

Evaluation of the Quality of Codonopsis Radix in Different Growth Years by the AHP-CRITIC Method.

The quality of traditional Chinese medicines (TCM) has a significant correlation with the source and growth years. However, there is no research on the relationship between the growth period and the quality of Codonopsis Radix (CR). This work aims to evaluate the quality of CR in different growth years (2-5 years). First, the content of 6 efficacy-related and 28 nutrient-related components in different growth years of CR was analyzed. The results showed that with the increase in growth years of CR, the content of some components increased, while some decreased. Then, the AHP-CRITIC method was performed to score the CR in different growth years, the results showed that the comprehensive score of CR increased with the increase of growth years, and the 5-year-CR had the highest score. Finally, in vitro activity assays were designed to verify the evaluation results. The results demonstrated that with the increase of growth years, the spleen lymphocyte proliferation activity and DPPH free radical scavenging activity of CR were enhanced, which proved that the AHP-CRITIC method is reasonable to evaluate the quality of CR in different growth years. The aforementioned findings demonstrated that CR quality improved with longer planting years.

Correction of Immunological and Behavioral Parameters of Rats with Experimental Traumatic Brain Injury with a Preparation of Monoclonal Antibodies to the C3 Component of Complement

After traumatic brain injury (TBI), inflammation develops in the CNS, an active participant in which is the complement system. Activated complement fragments initiate inflammation, and subsequently significantly affect the processes of repair and regeneration. The aim of the work is to reduce neuroimmune disorders after experimental TBI by blocking excessive inflammation in the early stages of traumatic disease with monoclonal antibodies to the C3 component of complement. The work was carried out on 65 male Wistar rats using the “falling weight” model. To correct neuroinflammation, a preparation of a recombinant monoclonal antibody 3A8, specific for the C3 neodeterminant of the rat complement component, blocking the activation of the alternative complement pathway was administered (i.v., 100 mg/kg). As a reference drug, a recombinant human interleukin-1 receptor antagonist (rIL-1RA) was used, which was administered s.c. (dose of 50 mg/kg). Both drugs were administered once after 30 min of TBI (mode 1) or 24 hours after TBI (mode 2). We studied the levels of corticosterone in the blood, the cytotoxic and proliferative activity of lymphocytes, and behavioral responses in the “plus maze” test. The obtained data indicate that on the 7th day after TBI in rats treated with 3A8 antibodies in mode 1, post-traumatic weight loss was decreased, the natural cytotoxicity of splenocytes and their proliferative activity were increased, and motor and exploratory activity were increased with a significant decrease in the level of anxiety. The introduction of rIL-1RA in these regimens, as well as the combined use of both drugs, did not have a significant effect on the studied parameters.

Preparation and anti-tumor effect of pig spleen ethanol extract against mouse S180 sarcoma cells in vivo

Numerous studies have shown that extracts derived from animal spleens have anti-tumor effects, but the biological activity of ethanol extract obtained from pig spleen has not been tested. Therefore, in the present study, we used ethanol as the extraction solvent to obtain the pig spleen ethanol extract (PSE) with immune activity, which provided a theoretical basis for further development and utilization of pig spleen as an anti-tumor drug. Then the structure characteristics and immune activity of PSE were investigated. According to the characterization analysis results, PSE was a high-purity protein complex mainly composed of hydrophobic amino acids, with an average molecular weight of 2.66 × 106 Da. The secondary structure was mainly α-helix and β-sheet. In vivo, PSE inhibited the growth of the S180 tumor, and immune organ toxicity was not detected. PSE induced S180 tumor cells to block and apoptosis in the S phase through the ROS-mediated mitochondrial pathway. Meanwhile, PSE improved the immunity of the body, such as promoting the proliferation activity of lymphocytes and increasing the level of IgG. In conclusion, PSE had significant cytotoxic and antiproliferative effects on S180 sarcoma cells.

Potential of Fig Leaf (Ficus carica ) as Immunomodulatory Agent in vitro and in Silico

Natural remedies are increasingly being used as alternatives to conventional medicine, and their popularity is growing. The fig plant (Ficus carica L.), which is known to contain secondary metabolites such as alkaloids, flavonoids, phenolics, terpenoids, steroids, and saponins, has the potential to act as an immunomodulator. The purpose of this study was to determine the content of flavonoid compounds based on Thin Layer Chromatography (TLC), total flavonoid content, total phenolic content, and immunomodulatory activity of ethanol extract and fig leaf fraction in vitro and silico (molecular docking). The fig leaf powder was extracted with 70% ethanol and fractionated with n-hexane and ethyl acetate to obtain ethanol extract, ethyl acetate fraction, n-hexane fraction, and water fraction of fig leaves. The extracts and fractions were then identified using the TLC method, and the total flavonoid and total phenolic levels were measured using the colorimetric method. Furthermore, the proliferation of lymphocyte cells and macrophage phagocytic activity were used to gauge the in vitro immunomodulatory activity. The results showed that the ethyl acetate fraction sample contained the highest total flavonoid and total phenolic content, namely 3.4630±0.04 mgQE/g sample and 220.1801±0.604 mgGAE/g sample. The immunomodulatory activity test findings revealed that fig leaf extract and fraction might enhance macrophage phagocytic activity in comparison to control cells. In the lymphocyte proliferation test, the value of IS>3 in the ethyl acetate fraction indicates that it has lymphocyte proliferation activity. This study showed that fig leaf extract and fraction could increase the phagocytic activity of macrophage cells. The ethyl acetate fraction of fig leaf could increase lymphocyte cell proliferation in vitro and silico.

PATHOLOGICAL IMMUNITY CHANGES IN COMORBID PATIENTS WITH EXACERBATION OF BRONCHIAL ASTHMA

The aim of the study – to investigate immunological changes in patients with exacerbation of bronchial asthma with different severity and controllability depending on the presence of comorbid pathology. Materials and methods of research. The work is based on the analysis of data from a comprehensive clinical and laboratory examination of 190 patients with exacerbation of asthma. The relative content of T–lymphocytes and their main subpopulations was determined on a flow cytofluorimeter FACS Calibur (Canada) by phenotyping lymphocytes with monoclonal antibodies (BECKMAN COULTER, USA) to surface membrane differentiation antigens (CD): CD3+19– (pan T-cells), CD4+–8+– (T−helpers/inducers), CD4–8+ (cytotoxic T lymphocytes), CD3–16+ (natural killers). To identify the imbalance of immunoregulatory subpopulations of T cells, the immunoregulatory index was calculated – the ratio of the number of CD4+8– Lf and CD4-8+ Lf. To calculate the absolute content in the peripheral blood of individual populations of Lf, leukogram indicators determined on the hematological analyzer ABH-miscros60, France were used. The proliferative activity of lymphocytes was evaluated in cultures of whole blood in the reaction of blast transformation of these cells with morphological accounting of the results. Statistical processing of the obtained data was carried out using licensed software products included in the Microsoft Office Professional 2007 package, license Russian Academic OPEN No Level № 43437596 in Excel. Conclusions. In moderate asthma, the frequency of pathological changes in the T-system of immunity did not depend on the presence of comorbid pathology, as a rule. In severe asthma, an increase in the content of pan–T cells occurred 1.2 times more often than in moderate asthma, but this was not observed in people with concomitant pathology. In this group, the frequency of increasing the content of T–helpers and decreasing the content of cytotoxic T – cells in patients with severe asthma without concomitant diseases was 1.3 times more frequent than in patients with comorbid pathology. No pathological changes in the T–system were detected in patients with controlled asthma. In partially controlled asthma, these changes were registered 1.5 times more often than in controlled asthma, and in its uncontrolled course, the prevalence of pathological changes in the T–system of immunity was much higher than in the two previous groups. In patients with concomitant diseases, these changes, as a rule, had a higher frequency than in patients without comorbid pathology.

In Vitro Immunomodulatory Activity of Fig Fruit Ethanol Extract (Ficus carica Linn) against Phagocytosis Macrophages and Lymphocyte Proliferation

Fig (Ficus carica L.) is a natural product that potentially can improve the immune system because it has flavonoids that have the potential as immunostimulants. The research aims to determine the possibility of fig fruit ethanol extract as an immunomodulator. Immunomodulatory activity is determined by knowing the activity of macrophage phagocytosis and lymphocyte proliferation in vitro and the levels of flavonoids in the extract. The research began with extraction, and then the sample was tested with TLC and colorimetry methods. Furthermore, the sample in the immunomodulatory activity test in vitro was measured through the activity of macrophage phagocytosis and lymphocyte proliferation. In the phagocytosis activity test, macrophage cells were given samples in various concentrations and latex beads. The number of activated macrophages and the number of latex phagocyted by the macrophage is then calculated. For tests of lymphocyte proliferation activity, lymphocyte cells were sampled with different concentrations and induced hepatitis B vaccine. Then the cell absorbance was read with an Elisa reader at 550nm wavelength. The study results found that the samples contained flavonoid compounds, and the total flavonoid levels obtained were 0.74±0.01 mgEQ/g samples. The immunomodulatory activity showed that the sample increased phagocytosis activity of macrophages compared to cell control. The lymphocyte proliferation test produced stimulation index&lt;2 values, showing no effect on the proliferation of lymphocytes. This study indicated that fig fruit ethanol extract could increase the phagocytosis activity of macrophage cells but did not affect the proliferation of lymphocyte cells in vitro.

Salvia miltiorrhiza polysaccharide activated macrophages and improved the disease resistance of sturgeon against Aeromonas hydrophila

The use of plant polysaccharides in aquaculture is recognized as a healthy strategy to enhance disease resistance and reduce medication use. Salvia miltiorrhiza polysaccharide (SMP) can regulate the immune function of higher vertebrates. However, the effects of SMP on fish have not been fully investigated. In this study, the ability of SMP to activate the macrophages of Siberian sturgeon (Acipenser bareii) was analyzed in vitro. The effects of SMP on immune cell activity of hybrid sturgeon (A. baerii ♀ × Acipenser schrenckii ♂) and resistance to Aeromonas hydrophila were further detected in vivo. The in vitro results showed that SMP up-regulated phagocytosis, respiratory burst, inducible nitric oxide synthase activity, nitric oxide (NO) concentration, and cytokine mRNA expression of macrophages. The in vivo results showed that dietary supplementation with SMP enhanced the respiratory burst of macrophages and proliferative activity of lymphocytes. Dietary supplementation with SMP increased serum concentrations of lysozyme and NO, and improved the survival rate of hybrid sturgeon challenged with A. hydrophila. Collectively, these results suggest that SMP can improve the immune function and disease resistance of sturgeon. This study provides a theoretical basis for the application of SMP for healthy farming of sturgeon.

Immunomodulatory and Anticancer Activities of Barley Bran Grown in Jordan: An in vitro and in vivo Study.

The Mediterranean diet is regarded as one of the most healthful dietary patterns in the world, owing to a combination of foods high in antioxidants and anticancer constituents. Barley bran is one of the components of the Mediterranean diet. It has nutritional and beneficial effects in different pathological conditions. Many studies were achieved to assess the nutritious values of barley bran, but there is no research indicating immunomodulatory and anticancer activities of barley bran grown in Jordan. The present study aims to examine and assess the potential immunomodulatory and anti-tumor activities of ethanol, n-hexane, aqueous/methanol, and water extracts obtained from barley bran. The Maceration method was utilized to prepare ethanol, n-hexane, aqueous/methanol, and water extracts. Various phytochemical groups were determined by using qualitative phytochemical tests. The antiproliferative activity of extracts was determined against MCF-7, HCT-116, A549, and EMT6/p by the MTT assay. The Folin-Ciocalteu reagent was used to detect the total phenolic content in extracts. Furthermore, immunomodulatory activity was assessed by determining the effect of extracts on splenocytes proliferation in the presence and absence of mitogens. The nitro blue tetrazolium assay and the neutral red method were used to assess the effect of each extract on the phagocytic activity of macrophages and pinocytosis, respectively. For the in vivo part, three different concentrations (10, 20, and 30% w/v) of barley bran were used to test the prophylactic effect in four Balb/C mice groups inoculated with EMT6/p cell-line subcutaneously. Also, serum samples were collected to assess the effect on cytokines (IFN-gamma, IL-2, IL-4, and IL-10). Barley bran extracts inhibited cancer cell proliferation. According to immunoassays, n-hexane and aqueous/methanol extracts could significantly rise lymphocyte proliferation and pinocytosis activity of macrophages. The activity of phagocytosis was increased by n-hexane and ethanol extracts. For the in vivo part, the average tumor size and weight of mice given the 30% barley bran group was significantly reduced (p < 0.05) compared with the control group. During our study, higher levels of TH1 cytokines (IFN- γ, IL-2) and lower levels of TH2 cytokine (IL-4) and T regulatory cytokine (IL-10) were obtained due to consumption of barley bran in food. Barley bran can be used as a prophylactic agent because it has anti-cancer and immunomodulatory activities.

Phytochemical Constituents of Propolis Flavonoid, Immunological Enhancement, and Anti-porcine Parvovirus Activities Isolated From Propolis

Propolis is widely used in health preservation and disease healing; it contains many ingredients. The previous study had revealed that the ethanolic or water extracts of propolis have a wide range of efficacy, such as antiviral, immune enhancement, anti-inflammatory, and so on, but its antiviral components and underlying mechanism of action remain unknown. In this study, we investigated the chemical composition, anti-porcine parvovirus (PPV) effectiveness, and immunological enhancement of propolis flavone ethanolic extracts. The chemical composition of propolis flavone was distinguished by ultra-performance liquid chromatography-quadrupole/time-of-flight tandem mass spectrometry analysis. In this study, the presence and characterization of 26 major components were distinguished in negative ionization modes to evaluate the effects of propolis flavonoid used as an adjuvant on the immune response of Landrace–Yorkshire hybrid sows immunized with an inactivated vaccine of PPV. Thirty Landrace-Yorkshire hybrid sows were randomly assigned to one of three groups, and the sows in the adjuvant groups were intramuscularly injected with PPV vaccine with a 2.0-ml propolis flavonoid adjuvant (PA) and oil emulsion adjuvant. After that, serum hemagglutination inhibition antibody titers and specific immunoglobulin (Ig)M and IgG subclasses were measured to evaluate the adjuvant effects of propolis flavonoid on the humoral immune responses, as well as peripheral lymphocyte proliferation activity and serum concentrations of Th1 and Th2 cytokines for cellular immunity. Results indicated an enhancing effect of PA on IgM, interleukins 2 and 4, interferon-γ, and IgG subclass responses. Especially in the effect of improving cellular immune response, the PA was the best. These findings suggested that PA can significantly enhance the immune responses against the PPV vaccine and could be an alternative way to improve PPV vaccination in sows. Furthermore, we screened the PF chemical components to the effectiveness of anti-PPV. Ferulic acid has an excellent anti-PPV effect.

Effects of Dietary Astragalus Polysaccharide Supplementation on the Th17/Treg Balance and the Gut Microbiota of Broiler Chickens Challenged With Necrotic Enteritis.

This study aimed to investigate the effects of dietary astragalus polysaccharide (APS) supplementation on the immune function, gut microbiota and metabolism of broiler chickens challenged with necrotic enteritis (NE). Two hundred forty Arbor Acres broiler chicks (one day old) were randomly assigned using a 2 × 2 factorial arrangement into two groups fed different levels of dietary APS (0 or 200 ppm of diet) and two disease challenge groups (control or NE challenged). The results showed that NE infection significantly increased FCR, mortality rate, Th17/Treg (Th17 cells% in blood and ileum, Th17/Treg, IL-17 and IL-17/IL-10 in blood), NO, lysozyme activity and IL-1β in blood, intestinal immune cell proportion and activity (Tc%, Treg% and monocyte phagocytic activity in ileum), intestinal inflammatory cytokines (TLR2, NF-κB, TNF-α and IL- 6) gene expression levels, and the number of Clostridium perfringens in cecum. NE infection significantly reduced body weight gain, thymus index, lymphocyte proliferation activity in blood and ileum, villus height and V/C in jejunum, Th cells% and Mucin2 gene expression in ileum. Dietary APS supplementation significantly increased body weight, feed intake, proportion of immune cells (T cells in blood and Tc, Treg in ileum), lymphocyte proliferation activity, V/C in jejunum, and ZO-1 gene expression in ileum. Dietary APS supplementation significantly reduced FCR and mortality rate, Th17/Treg, Th17%, intestinal pathology scores, intestinal inflammatory cytokine gene expression levels, and the number of Clostridium perfringens in cecum. In addition, broilers challenged with NE significantly increased Staphylococcus and Turicibacter and reduced α diversity of microbiota in ileum. Dietary APS supplementation significantly increased α diversity, Romboutsia, Halomonas, propionic acid, butyric acid, formononetin, taurine, cholic acid and equol and downregulated uric acid, L-arginine and serotonin in ileum. Spearman’s correlation analysis revealed that Romboutsia, Turicibacter, Staphylocpccus, Halomonas, Streptococcus, Escherichia-Shigella, Prevotella, uric acid, L-arginine, jerivne, sodium cholate and cholic acid were related to inflammation and Th17/Treg balance. In conclusion, APS alleviated intestinal inflammation in broilers challenged with NE probably by regulating intestinal immune, Th17/Treg balance, as well as intestinal microbiota and metabolites.

Immunostimulatory and antioxidant activities of the selenized polysaccharide from edible Grifola frondosa.

Grifola frondosa polysaccharide (GFP2) was extracted and purified by anion‐exchange chromatography. A selenized G. frondosa polysaccharide, SeGFP2, was modified in selenylation by nitric acid–sodium selenite (HNO3‐Na2SeO3) method. Structural features were investigated, and the lymphocyte proliferation and antioxidant activities were compared taking GFP2 as control. SeGFP2 with a molecular weight of 2.12 × 104 Da was composed of mannose, glucose, and galactose with a ratio of 3.5:11.8:1.0. A typical absorption of selenium ester was observed in SeGFP2 molecule. SeGFP2 was proposed as a branched polysaccharide, which consisted of 1,3‐D‐Glcp, 1,6‐D‐Glcp, 1,4,6‐D‐Galp, and 1,3,6‐D‐Manp. SeGFP2 showed a linear filamentous structure with some branches. SeGFP2 could significantly promote T‐ or B‐lymphocyte proliferation and the enhancement was higher than GFP2. The in vitro antioxidant activities of SeGFP2 were more potent than GFP2. These present data suggested that selenylation could significantly improve the lymphocyte proliferation and in vitro antioxidant activities of GFP2.