• Microbiology and Biotechnology Letters
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• v.42 no.3
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• pp.312-315
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• 2014

So as to evaluate its use an immune stimulator in humans, the toxicity and action against immune cells by an anti-yeast substance (AYS), a bacterial proteoglycan, were investigated. The AYS did not possess hemolytic activity with human red blood cells (hRBC). Nor did it exhibit cytotoxicity against human peripheral blood mononuclear cells (hPBMC). In addition, the AYS did not induce hPBMC proliferation, but it did agglutinate hPBMCs in vitro. Moreover, hPBMC induced inflammatory cytokines such as IL-6, IL-5, IFN-${\gamma}$ and TNF-${\alpha}$ with the AYS during culture. Compared with alum, the AYS as an adjuvant has an increased antibody production rate against bovine serum albumin (BSA) in mice.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.631-636
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• 2008

Acanthopanax divaricatus var. albeofructus (ADA) have been shown to have various levels of activity such as antioxidant, anticancer, antivirus, and immunostimulatory effects. However, little is known about its mechanism related to the modulation of immune activities. In this study, a water extract of ADA leaves were used to treat human peripheral blood mononuclear cells (hPBMC) to determine the underlying mechanisms for the immunostimulatory effects. To characterize its immunomodulatory activity, the secretion level of various cytokines including IL-2, IL-4, IL-6, IL-10, IL-12, IFN-$\gamma$, and TNF-$\alpha$ were measured using enzyme-linked immunosorbent assay (ELISA). Treatment of hPBMC with ADA leaf extract in an in vitro experiment induced various Th1 cytokines in a dose-dependent manner. A significant increase of IL-2, IL-12, IFN-$\gamma$, and TNF-$\alpha$ secretion was observed in the presence of ADA leaf extract. In contrast, Th2 cytokines including IL-4 and IL-6 were suppressed. There was no significant change in IL-10 release. Our results showed an increase in Th1 and a decrease in Th2 cytokine secretion which suggests that ADA may influence the immune response towards a predominance of Th1 cytokines in the immune system.

• Journal of Animal Science and Technology
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• v.49 no.2
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• pp.213-224
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• 2007

In this study, hatched male broiler chicks(Ross) were fed on a basal diet and LPS was administered via intraperitoneal injection three times every other day, on the 9th, 11th and 13th days of the experiment, and then PBMC and splenocytes were isolated on day 14. The degree of alama blue reduction was evaluated at 4, 24, 48, 96 and 120 h in the splenocytes, and at 4, 8, 12, 24 and 48 h for PBMC of incubation after the addition of alama blue solution to the media. The cell numbers used in this experiment were 103, 104 and 105 cells per well, and the con A levels were 0.0, 1.0, 5.0, and 10.0 ㎍ per ml of medium. 1. The degree of alama blue reduction was found to increase in a linear fashion with increasing incubation time and cell numbers, for both splenocytes and PBMC. 2. During acute phase response, the degree to which alama blue was reduced was significantly elevated (p<0.05) at an incubation time of 24 hr for the splenocytes, 4 hr for PBMC, and a cell number of 105 cells per well, respectively. 3. The raised reduction of alama blue to control was linear with Con A levels in medium, and higher reduction in Con A 10.0 ㎍ relative to 1.0 or 5.0 ㎍ in ml medium was shown 4. The medium with autologous serum evidenced a significantly (p<0.05) higher reduction of alama blue relative to FBS. 5. Splenocytes and PBMC from the LPS-injected birds evidenced significantly higher levels of alama blue reduction regardless of incubation time, number of cells, level of Con A added, or serum type, as compared with what was observed in normal birds. The results indicated that the assay conditions for proliferative activity using the alama blue method in birds in which the acute phase response had been activated via intraperitoneal LPS injection requires 4 hrs of incubation for PBMC, 24 hrs of incubation for splenocytes, and 10㎍ of Con A per ml of medium.

• Archives of Pharmacal Research
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• v.28 no.5
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• pp.573-581
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• 2005

Flavonoids, a group of low molecular weight phenylbenzopyrones, have various pharmacological properties including antioxidant activity, anticancer, and immunomodulatory effects. In the present study, lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate/phytohemagglutinin (PMA/PHA) were used as stimulants for RAW 264.7 macrophages and human peripheral blood mononuclear cell (hPBMC), and tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-2 productions were measured. In addition, flavonoids were examined for their effects on LPS-induced NO production in RAW 264.7 macrophages. The results showed that all compounds were not strongly cytotoxic at the tested concentrations on hPBMC and RAW 264.7 macrophages. On immunomodulatory properties, catechin, epigallocatechin (EGC), naringenin, and fisetin repressed NO production and TNF-${\alpha}$ secretion. Furthermore, catechin, epigallocatechin gallate (EGCG), epicatechin (EC), luteolin, chrysin, quercetin, and galangin increased IL-2 secretion while EGC, apigenin, and fisetin inhibited the secretion. These results indicated that flavonoids have the capacity to modulate the immune response and have a potential anti-inflammatory activity. There was no obvious structure-activity relationship regard to the chemical composition of the flavonoids and their cell biological effects.

• Korean Journal of Pharmacognosy
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• v.37 no.4 s.147
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• pp.221-228
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• 2006

A lectin was purified from Serrognathus platymelus castanicolor larvae and named as SPL. The purification was carried out by ion-exchange chromatography on DEAE Sephadex A-50 and gel filtration chromatography on Sephadex G-200. The purity of the protein was verified by polyacrylamide gel electrophoresis and the purified lectin agglutinated erythrocytes of rabbit and human A, B, O, AB. SPL was tested it's ability to enhance the expressions of cytokines, $IL-1\alpha$, IL-2, IL-6, $TNF\alpha$ and $IFN\gamma$ by human peripheral blood mononuclear cells (PBMC) obtained from healthy donors. mRNA analyses were performed by RT-PCR at the moment of 1, 4, 8, 24, 48, 72 and 96 h after stimulation of PBMC with purified SPL. The patterns of IL-2 band were slightly expressed from 24 h and the strongest band was appeared at 96 h. The expressions of $IL-1\alpha$ and IL-6 mRNA were strong from 1 to 8 h and those of $TNF\alpha$ were from 48 to 96 h. The mRNA encoding $IFN\gamma$ were not detected. The addition of SPL for macrophage cultures induced production of nitric oxide (NO) by cells in a dose-dependent manner. NO release was partially inhibited by $TNF\alpha$ antibodies. These results suggest that SPL has the ability to enhance cytokine expressions in PBMC and to induce the NO release by TNFa in macrophage cultures from PBMC cultures.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.8
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• pp.1182-1187
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• 2003

Two experiments involving 92 crossbred, 21 day old weaned pigs were used to evaluate the effect of glutamine supplement in a dietary or culture medium on lymphocyte proliferation. In Exp. 1, 88 pigs were fed diets supplemented with 0, 0.5, 1.0, or 1.5% glutamine for 28 days. Lymphocytes were prepared from peripheral blood mononuclear cells (PBMC), ileal Peyer's patches (PP), the mesenteric lymph node (MLN), and the spleen in each dietary supplement group on days 7, 14, or 28 postweaning. Lymphocytes were cultured at $37^{\circ}C$ for 72 h in a RPMI-1640 medium with or without mitogen-stimulated, and pulsed with 3Hthymidine for an additional 18 h. The stimulation index of PBMC proliferation in 1.0% dietary glutamine supplement group and both of the MLN and splenocytes proliferation in 1.5% dietary glutamine supplement group was significantly (p<0.05) increased at 14 days postweaning. In Exp. 2, four weaned pigs were fed a basal diet for 14 days. The 3H-thymidine incorporation of PBMC, PP, and MLN cells, incubated with 0.125 to 0.25 mM glutamine in culture medium were markedly enhanced with Con A-stimulated, however, the splenocyte proliferation was not affected in the addition of glutamine medium. These observations suggest that dietary glutamine supplement might enhance the lymphocyte proliferation of weaned pigs.

• YAKHAK HOEJI
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• v.43 no.4
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• pp.474-480
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• 1999

The purpose of this study is to define whether Asterina pectinifera Lectin (APL) is effective on the cytokine production. Isolated mRNA from hPBMC (human peripheral blood mononuclear cells) stimulated with APL for various reaction times (1 to 96 hours) was detected by RT-PCR. The intensity of band for IL-1 and $IFN{\gamma}$ mRNA was markedly increased at l hour, and IL-2 mRNA was strongly expressed at 4 hours. The mRNA band of APL-induced IL-2 and $IFN{\gamma}$ was weaker than that of IL-1, IL-6 and $TNF{\alpha}$. The mRNA expression of 4 cytokines (IL-1, IL-2, $IFN{\gamma}$ and $TNF{\alpha}$) was detected up to 48 hours, and that of IL-6 was detected until 72 hours. ELISA was used to look protein secretion of the cytokine gene with IL-1, IL-2 and TNF$\alpha$expressed strongly in RT-PCR. The highest protein secretion was at 4 hours with IL-1, at 8 hours with IL-2 and at 4 hours with $TNF{\alpha}$. These results suggest that APL can induce the production of some cytokines and the immune response from PBMC was done within the first few hours of stimulation with APL.

• Tuberculosis and Respiratory Diseases
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• v.44 no.3
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• pp.470-478
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• 1997

Background : Phagocytosis is probably the first step for mycobacteria to be virulent in host because virulent strains are more readily phagocytosed by macrophage than attenuated strains. According to the traditional concept, multi-drug resistant strains have been regarded as less virulent. However, this concept has been challenged, since recent studies(reported) showed that the degree of virulence and drug-resistance is not related. The purpose of this study is to evaluate whether the phagocytic activity of M.tuberculosis by peripheral blood mononuclear cells(PBMC) is different according to drug-resistance or host factor. To evaluate this, we estimated the difference of phagocytic activity of drug-resistant and drug-sensitive M.tuberculosis and also estimated the phagocytic activity of PBMC from intractable tuberculosis patients and healthy controls. Methods : PBMC from ten intractable tuberculosis patients and twelve healthy control, and three different strains of heat-killed M.tuberculosis, ie, ADS(all drug sensitive), MDR(multi-drug resistant), and ADR(all drug resistant) were used. After incubation of various strains of M.tuberculosis with PBMC, the phagocytic activity was evaluated by estimating proportion of PBMC which have phagocytosed M.tuberculosis. Results : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains(Percentage of PBMC phagocytosed M.tuberculosis in healthy control : ADS : $32.3{\pm}2.9%$, ADR : $49.6{\pm}3.4%$, p = 0.0022, Percentage of PBMC phagocytosed M.tuberculosis in intractable tuberculosis patients : ADS : $34.9{\pm}3.6%$, ADR : $50.7{\pm}4.5%$, p = 0.0069). However, there was no difference in phagocytic activity of PBMC from healthy control and intractable tuberculosis patients. Conclusion : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains and host factors does not seems to influence the phagocytosis of M.tuberculosis.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.13
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• pp.5343-5348
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• 2015

Promoter hypermethylation of the runt-related transcription factor 3 (RUNX3) gene is associated with increased risk of hepatocellular carcinoma (HCC). Oxidative stress plays a vital role in both carcinogenesis and progression of HCC. However, whether oxidative stress and RUNX3 hypermethylation in HCC have a cause-and-effect relationship is not known. In this study, plasma protein carbonyl and total antioxidant capacity (TAC) in patients with hepatitis B virus (HBV)-associated HCC (n=60) and age-matched healthy subjects (n=80) was determined. RUNX3 methylation in peripheral blood mononuclear cells (PBMC) of subjects was measured by methylation-specific PCR. Effect of reactive oxygen species (ROS) on induction of RUNX3 hypermethylation in HCC cells was investigated. Plasma protein carbonyl content was significantly higher, whereas plasma TAC was significantly lower, in HCC patients than healthy controls. Based on logistic regression, increased plasma protein carbonyl and decreased plasma TAC were independently associated with increased risk for HCC. PBMC RUNX3 methylation in the patient group was significantly greater than in the healthy group. RUNX3 methylation in hydrogen peroxide ($H_2O_2$)-treated HepG2 cells was significantly higher than in untreated control cells. In conclusion, increase in oxidative stress in Thai patients with HBV-associated HCC was demonstrated. This oxidative increment was independently associated with an increased risk for HCC development. RUNX3 in PBMC was found to be hypermethylated in the HCC patients. In vitro, RUNX3 hypermethylation was experimentally induced by $H_2O_2$. Our findings suggest that oxidative stress is a cause of RUNX3 promoter hypermethylation in HCC cells.

• Animal Bioscience
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• v.36 no.7
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• pp.1143-1149
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• 2023

Objective: We investigated the effects of outdoor access for 1 h per day on the animal welfare (AW) of tethered cows, in terms of lying and sleeping postures, and immune function. Methods: A total of five dry cows were tethered all day indoors (tethering) for 30 days and then tethered indoors with 1 h daily outdoor access (ODA-1h) for 30 days. To analyze the effects of ODA-1h, we calculated the total duration and bout frequency per day, and bout duration of lying and sleeping postures during the last five days of each treatment period. We also analyzed the populations of T cells, B cells, and NK cells in peripheral blood mononuclear cells (PBMC) by fluorescence-activated cell sorting and determined the concanavalin A (Con A) -induced proliferation rate of T cells. Results: The mean total time per day of lying during the ODA-1h treatment was significantly shorter than that during the tethering treatment (p<0.001). The Con A-induced proliferation rate of T cells during the ODA-1h treatment was significantly higher than that during the tethering treatment (p = 0.007). The proportion of NK cells in PBMC during the ODA-1h treatment tended to be higher than that during the tethering treatment (p = 0.062). Conclusion: Although ODA-1h may decrease lying time, it increases the available space for tethered cows towards that typically found in grazing and free barn feeding systems. This increased available space promotes the expression of normal behaviors such as walking and social behaviors except lying and may also improve the immune function of tethered dry cows, thereby improving their overall welfare.