• Proceedings of the Korea Society of Poultry Science Conference
• /
• 2003.11a
• /
• pp.82-83
• /
• 2003

To study effect of bean extracts to lessen the growth-suppressing-effect of krill meal diet, dietary krill meal with bean extracts on the performance of broiler chicks and proliferation of splenocytes and peripheral blood mononuclear cells(PBMC) and levels of circulating TNF-$\alpha$ and ovotransferrin in plasma was assayed. The krill meal with bean extracts diet lessened the growth-suppressing effect of the krill meal diet. During acute phase responce, the krill meal with bean extracts diet decreased the proliferation of splenocytes and increased the proliferation of the PBMC and reduced the circulating levels of TNF-$\alpha$ and ovotransferrin in plasma. The results Indicated that the krill meal with bean extracts diet related with the acute phase response in broiler chicks.

• Proceedings of the Korea Society of Poultry Science Conference
• /
• 2005.11a
• /
• pp.76-77
• /
• 2005

Effects of dietary killed salmonella and vancomycin on the performance and immune response was investigated in broiler chicks inoculated with Salmonella typhimurium. During 3 week(27 d) of age, experimental diet did not affect daily gain, feed intake and feed efficiency. Dietary killed salmonella and vancomycin decreased MnSOD activity. At 7day after Salmonella typhimurium inoculation, dietary killed salmonella and vancomycin increased, daily gain and feed efficiency of broiler chicks. At 7 and 15 day after salmonella inoculation, dietary killed salmonella and vancomycin decreased erythrocyte peroxidase activity, but elevated proliferation of PBMC stimulated with LPS and supernatant IL-1 level secreted by the PBMC. The results suggested that dietary killed salmonella and vancomycin improved the performance of broiler chicks due to modulate antioxidant system and innate immune response of broiler chicks innoculated with Salmonella typhimurium.

• Korean Journal of Veterinary Research
• /
• v.42 no.2
• /
• pp.209-217
• /
• 2002

This study was performed to produce monoclonal antibodies (mAb) specifically reacting with chicken leukocyte surface antigens. Popliteal lymph node cells of BALB/c mice previously immunized through foot-pad with peripheral blood mononuclear cells (PBMC) of chickens separated by Ficoll-Histopaque method. They were fused with P3X63Ag14 mouse myeloma cells. A total of 34 hybridomas secreted antibodies specifically binding to the PBMC. According to the reactivity patterns with PBMC, the mAbs were divided into 4 groups. Group 1 mAbs (IIB3, IIB10, IIE10) specifically reacted with non-adherent lymphocytes but not with adherent cells which were mainly composed of thrombocytes and monocytes in PBMC culture. These mAbs were reactive with 25-59% of thymus cells and 42-64% of spleen cells of chickens. They did not show any significant reactivity with cells in the bursa of Fabricius, T-cell (MDCC-MSB1) and B-cell (LSCC-1104B1) lines. These results indicate that Group I mAbs specifically reacted with T-lymphocyte subpopulation. Monoclonal antibodies in Group II (IC6, IG2-2 and IID9) showed specific reactivity with monocytes but not with thrombocytes or non-adherent cells in PBMC culture. These mAbs, though not reacted with the chicken macrophage cell line, HD11, also bound to macrophages of the spleen and lung in immunohistochemical staining. Five mAbs in Group III showed characteristics of binding to lymphocytes and monocytes, but not to thrombocytes. Twenty-three mAbs in Group IV showed specific reactivity to lymphocytes, monocytes, and thrombocytes. Two mAbs (IC3 and IE9) in Group IV reacted with most of PBMC.

• Asian-Australasian Journal of Animal Sciences
• /
• v.27 no.4
• /
• pp.471-478
• /
• 2014

Investigating gene expression of immune cells of whole blood or peripheral blood mononuclear cells (PBMC) under polyinosinic:polycytidylic acid (poly I:C) stimulation is valuable for understanding the immune response of organism to RNA viruses. Quantitative real-time PCR (qRT-PCR) is a standard method for quantification of gene expression studies. However, the reliability of qRT-PCR data critically depends on proper selection of reference genes. In the study, using two different analysis programs, geNorm and NormFinder, we systematically evaluated the gene expression stability of six candidate reference genes (GAPDH, ACTB, B2M, RPL4, TBP, and PPIA) in samples of whole blood and PBMC with or without poly I:C stimulation. Generally, the six candidate genes performed a similar trend of expression stability in the samples of whole blood and PBMC, but more stably expressed in whole blood than in PBMC. geNorm ranked B2M and PPIA as the best combination for gene expression normalization, while according to NormFinder, TBP was ranked as the most stable reference gene, followed by B2M and PPIA. Comprehensively considering the results from the two programs, we recommended using the geometric mean of the three genes, TBP, PPIA and B2M, to normalize the gene expression of whole blood and PBMC with poly I:C stimulation. Our study is the first detailed survey of the gene expression stability in whole blood and PBMC with or without poly I:C stimulation and should be helpful for investigating the molecular mechanism involved in porcine whole blood and PBMC in response to poly I:C stimulation.

• Tuberculosis and Respiratory Diseases
• /
• v.57 no.4
• /
• pp.336-344
• /
• 2004

Background : Immunotherapy for cancer has not been successful because of several obstacles in tumor and its environment. Inappropriate secretions of cytokines and growth factors by tumors cause substantial changes in the immune responses against tumors, affording the tumors some degree of protection from immune attack. Uteroglobin (UG, Clara cell secretory protein) has been known to have anti-inflammatory, immunomodulatory and anti-cancer activities. However, in lung cancer cells, UG expression is decreased. This study investigated the role of UG in the immunomodulation of lung cancer. Methods : The UG protein was overexpressed by Adenovirus(Ad)-UG transduction in non-small cell lung cancer cell lines. The concentration of Prostaglandin $E_2$ ($PGE_2$) was measured by Enzyme Immunoassay (EIA). Peripheral blood mononuclear cells (PBMC) from whole blood were prepared with Ficoll. PBMC were cultured in RPMI 1640, supernatant of A549, or A549 with UG or NS-398. Concentration of Th 1 type and Th 2 type cytokines from PBMC were measured by ELISA. Results : UG suppressed $PGE_2$, Cyclooxygenase-2 (COX-2) product. Both Th1 type such as Interleukin-2 (IL-2), Interferon-${\gamma}$ (IFN-${\gamma}$) and Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and Th2 type cytokines such as IL-10 and Tumor growth factor-${\beta}$ (TGF-${\beta}$) were increased when PBMC were cultured with supernatant of non small lung cancer cells. UG and COX-2 inhibitor, NS-398 induced normal immune response of PBMC. Although Th 1 type cytokines were increased, Th 2 type cytokines were reduced by UG. Conclusion : UG suppressed PGE2, COX-2 product. Supernatant of NSCLC induced imbalance of immune response of PBMC. However, UG reversed this imbalance. These results suggest that UG may be used in the development of immunotherapy for lung cancer.

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

• The Journal of Korean Medicine
• /
• v.22 no.1
• /
• pp.53-62
• /
• 2001

Objective : The aim of this study was to evaluate the efficacy of Injinchunggan-tang on Hepatitis C virus infection, and to clarify the mechanism of treatment by indentifying the effect of Injinchunggan-tang on cytokine secretion. Methods : In vitro model of HCV infection in MOLT 4 cell was used. The effect of Injinchunggan-tang on the attachment of HCV on MOLT 4 cell was studied by PCR method. The change of cytokine secretion according to Injinchunggan-tang treatment was investigated by ELISA. Results : Injinchunggan-tang inhibited the attachment of HCV on MOLT 4 in the concentration of $10-2{\mu\textrm{g}}/\mu\textrm{\ell}$ and $10-1{\mu\textrm{g}}/\mu\textrm{\ell}$. In cytokine assay, Injinchunggan-tang increased the secretion of IL-4 of mouse splenocytes and PBMC in 48 hour culture as well as the secretion of IL-12 of mouse splenocytes and PBMC in 48 hour culture, whereas it decreased the secretion of $IFN-{\gamma}$ of mouse splenocytes in 24 and 48 hour culture. Conclusion : The results of this study show that Injinchunggan-tang has an inhibitory effect on the attachment on HCV on Mo1t4 Cell, and that it increases the secretion of IL-4 and IL-12 of mouse splenocyte and PBMC.

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

• Proceedings of the Korea Society of Poultry Science Conference
• /
• 2005.11a
• /
• pp.78-79
• /
• 2005

Effects of dietary lactose or killed Salmonella on the performance, immune response and anti-oxidant system was studied in chicks innoculated with Salmonella typhimurium. In 27 days of age broiler, dietary lactose decreased performance, while dietary lactose and killed Salmonella elevated plasma peroxidase activity and IL-1 level in supernatant of PBMC stimulated with LPS. When broiler chicks innoculated with Salmonella, performance, activities of erythrocyte MnSOD and plasma peroxidase were enhanced after 7 days of the innoculation. Dietary lactose and killed Salmonella increased activity of erythrocyte MnSOD, plasma peroxidase, proliferation of PBMC stimulated with LPS and IL-1 level in the supernatant after 15 days of the innoculation. The result indicated that dietary lactose and killed Salmonella have modulated innate immune response and antioxidant system in broiler chicks innoculated with Salmonella typhimurium.

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