• 약학회지
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• 제51권4호
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• pp.270-276
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• 2007

Silybin is known to be a major active flavonoid component isolated from Silybum marianum, a hepatoprotective medicinal plant. In this study, we examined the immunomodulatory role of silybin on T cell and macrophage-mediated immune responses. To do this, the proliferation of splenic lymphocytes and CD8+ CTLL-2 cells under mitogenic stimulation with lipopolysaccharide (LPS), concanavalin (Con) A and interleukin (IL)-2 and the production of $TNF-{\alpha}$ and NO from LPS- and $IFN-{\gamma}$-activated macrophages was evaluated under silybin treatment. The mitogenic proliferation of splenic lymphocytes induced by LPS and Con A was strongly diminished by silybin in a dose-dependent manner. Moreover, the proliferation of CD8+ CTLL-2 cells was also negatively modulated by the compound. In contrast, silybin did not strongly suppress the proliferation of normal splenocytes and T cell line Sup-T1 cells, indicating that the inhibitory effect of silybin may be due to blocking only mitogenic responses of splenic lymphocytes. In addition, silybin inhibited $TNF-{\alpha}$ production in LPS-stimulated RAW264.7 cells. Effect of silybin however was distinct, according to NO-inducing stimuli. Thus, silybin only blocked NO production induced by $IFN-{\gamma}$ but not LPS and the inhibition was increased when PMA was co-treated with $IFN-{\gamma}$. Unlike NO inhibition, however, this compound protected the cytotoxic damage of RAW264.7 cells induced by both LPS and $IFN-{\gamma}$. Therefore, our data suggest that silybin may participate in host immune responses mediated by T cells and macrophages via regulating mitogenic proliferation, and the production of $TNF-{\alpha}$ and NO, depending on cellular stimuli.

• Archives of Pharmacal Research
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• 제23권3호
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• pp.240-242
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• 2000

The immunomodulating activity of a polysaccharide isolated from Morus alba (PMA) root bark was examined in murine splenic lymphocytes. PMA enhanced proliferation of splenic lymphocytes in a synergistic manner in the presence of mitogens. However, PMA suppressed pri-may IgM antibody production from B cells, which was activated with lipopolysaccharide, a polyclonal activator, or immunized with a T-cell dependent antigen sheep red blood cells. Our observations showed that the immunomodulating activity of PMA increased lymphocyte proliferation and that PMA decreased antibody production from B cells, which was distinct from those of other plant-originated polysaccharides.

• 한국식품영양과학회지
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• 제26권6호
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• pp.1194-1199
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• 1997

The effect of hot taste preference on selected immune responses was investigated in human peripheral immunocompetent cells. Human lymphocytes and natural killer(NK) cells were prepared at a concentration of 2$\times$$10^{6}$ cells/ml in RPMI-1640 containing 10% fetal bovine serum. Lymphocytes proliferation was determined with the [$^{3}H$]-thymidine pulse for 18hrs after concanavalin A, phytohemagglutinin, Salmonella typhimurium mitogen, or media alone. NK cell activity was measured by cytolysis of $^51Cr$-labeled target cells K562. Serum antibodies levels such as IgM, IgG, IgA were also measured by ELISA method. There was no difference of serum IgM level among the groups, but IgG and IgA levels were greater in the group with hot taste preference than those of the group without hot taste preference. In lymphocytes of the group with hot taste preference there was a greater mitogen-induced lymphocyte proliferative responses compared to the group without hot taste preference. In addition, NK cell activity in group with hot taste preference was lower than that of the group without hot taste preference. These results suggest that the eating habit of spicy food containing hot components may affect immune status by modulating selective immunocompetent cells function.

• Asian Pacific Journal of Cancer Prevention
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• 제15권3호
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• pp.1119-1123
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• 2014

Head and neck cancers (HNC) are extremely complex disease types and it is likely that chromosomal instability is involved in the genetic mechanisms of its genesis. However, there is little information regarding the background levels of chromosome instability in these patients. In this pilot study, we examined spontaneous chromosome instability in short-term lymphocyte cultures (72 hours) from 72 study subjects - 36 newly diagnosed HNC squamous cell carcinoma patients and 36 healthy ethnic controls. We estimated chromosome instability (CIN) using chromosomal aberration (CA) analysis and nuclear level anomalies using the Cytokinesis Block Micronucleus Cytome Assay (CBMN Cyt Assay). The proliferation rates in cultures of peripheral blood lymphocytes (PBL) were assessed by calculating the Cytokinesis Block Proliferation Index (CBPI). Our results showed a significantly higher mean level of spontaneous chromosome type aberrations (CSAs), chromatid type aberration (CTAs) dicentric chromosomes (DIC) and chromosome aneuploidy (CANE UP) in patients (CSAs, $0.0294{\pm}0.0038$; CTAs, $0.0925{\pm}0.0060$; DICs, $0.0213{\pm}0.0003$; and CANE UPs, $0.0308{\pm}0.0035$) compared to controls (CSAs, $0.0005{\pm}0.0003$; CTAs, $0.0058{\pm}0.0015$; DICs, $0.0005{\pm}0.0003$; and CANEUPs, $0.0052{\pm}0.0013$) where p<0.001l. Similarly, spontaneous nuclear anomalies showed significantly higher mean level of micronuclei (MNi), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) among cases (MNi, $0.01867{\pm}0.00108$; NPBs, $0.0156{\pm}0.00234$; NBUDs, $0.00658{\pm}0.00068$) compared with controls (MNi, $0.00027{\pm}0.00009$; NPBs, $0.00002{\pm}0.00002$; NBUDs, $0.00011{\pm}0.00007$).The evaluation of CBPI supported genomic instability in the peripheral blood lymphocytes showing a significantly lower proliferation rate in HNC patients ($1.525{\pm}0.005552$) compared to healthy subjects ($1.686{\pm}0.009520$) (p<0.0001). In conclusion, our preliminary results showed that visible spontaneous genomic instability and low rate proliferation in the cultured peripheral lymphocytes of solid tumors could be biomarkers to predict malignancy in early stages.

• Journal of Ginseng Research
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• 제33권1호
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• pp.33-39
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• 2009

각 체질별(태음인, 소양인, 소음인)로 5명씩 혈액을 채취하여 임파구를 분리한 후 백삼 및 홍삼 에탄을 추출물을 투여하여 면역효능(proliferation, NO생성 및 TNF-$\alpha$의 분비능)을 측정한 결과, 한방에서 음인을 보하는 약재로 알려진 백삼의 경우 소음인 군의 임파구에서 가장 높은 임파구 증식능, NO 생성 및 TNF-$\alpha$의 분비능을 나타냈으나, 소양인군에서는 가장 낮은 면역활성을 나타내었다(P<0.05). 그러나 홍삼 추출물의 경우 각 체질군에서 면역활성의 차이가 없이 유사한 것으로 나타났다(P > 0.05). 또한, 체질별 임파구에 LPS처리한 대조군보다 백삼 및 홍삼 추출물을 첨가한 처리군에서 NO생성 및 TNF-$\alpha$의 분비능이 현저하게 높은 것으로 나타났다(P<0.05).백삼과 달리 홍삼 추출물에서 각 체질군에 면역활성의 차이가 없는 것은 인삼의 가공처리 과정에서 성분변화의 차이에 기인하는 것으로 사료된다. 따라서 본 연구 결과, 홍삼추출물로부터 체질에 영향을 미치지 않는 성분을 규명한다면 새로운 맞춤형 체질마커로 개발할 수 있으리라 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제14권8호
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• pp.1179-1187
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• 2001

In fish both humoral and cell mediated immune responses have been reported whereas antibodies recognizing specific cellular populations have not yet been developed except for ones recognizing surface Ig molecules on B lymphocytes. Our aim was to develop and characterize monoclonal antibodies (Mabs) specific for the immune-related cells. Mabs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously sensitized against Israeli carp (I. carp) kidney mononuclear cells. We obtained 44 Mabs positively reacting with I. carp kidney mononuclear cells and partially characterized 7 Mabs in the morphological and mitogen-based proliferative aspects. Fluorescence-activated cell sorter (FACS) analysis against I. carp kidney cells by using 7 different Mabs showed 80.3% for ICK 17-4, 65.1% for ICK 2-3, 64.1% for ICK 25-1, 67.5% for lCK 22-1, 70.8% for ICK 16-2, 76.8% for ICK 13-2, 79.7% for ICK II-I. Panning method was used for the isolation of Mabs specific mononuclear carp spleen cells followed by Wright's stain. The stained cell populations were identified as monocytes (ICK 17-4, ICK 2-3, ICK 25-1, ICK 22-1 and ICK 16-2), lymphocytes (ICK 11-1), and a mixed cell population of monocytes and lymphocytes (ICK 13-2). In cell proliferation assay, monocytes purified by ICK 17-4, 2-3 and 22-1 efficiently responded to Con A and PHA, while ones separated by ICK 25-1 did not react with any mitogens. Lymphocytes isolated by ICK 11-1, though it is not known whether they are T or B cells, were more responsive to Con A than PHA or LPS, suggesting that fish immune cells are somewhat different from mammalian cells in responding to mammalian T or B cell mitogens.

• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.611-616
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• 2014

Objectives: Dendritic cell (DC)-based tumor immunotherapy needs an immunogenic tumor associated antigen (TAA) and an effective approach for its presentation to lymphocytes. In this study we explored whether transduction of DCs with lentiviruses (LVs) expressing the human interleukin-12 gene could stimulate antigen-specific cytotoxic T cells (CTLs) against human lung cancer cells in vitro. Methods: Peripheral blood monocyte-derived DCs were transduced with a lentiviral vector encoding human IL-12 gene (LV-12). The anticipated target of the human IL-12 gene was detected by RT-PCR. The concentration of IL-12 in the culture supernatant of DCs was measured by ELISA.Transduction efficiencies and CD83 phenotypes of DCs were assessed by flow cytometry. DCs were pulsed with tumor antigen of lung cancer cells (DC+Ag) and transduced with LV-12 (DC-LV-12+Ag). Stimulation of T lymphocyte proliferation by DCs and activation of cytotoxic T-lymphocytes (CTL) stimulated by LV-12 transduced DCs pulsed with tumor antigen against A549 lung cancer cells were assessed with methyl thiazolyltetrazolium (MTT). Results: A recombinant lentivirus expressing the IL-12 gene was successfully constructed. DC transduced with LV-12 produced higher levels of IL-12 and expressed higher levels of CD83 than non-transduced. The DC modified by interleukin -12 gene and pulsed with tumor antigen demonstrated good stimulation of lymphocyte proliferation, induction of antigen-specific cytotoxic T lymphocytes and antitumor effects. Conclusions: Dendritic cells transduced with a lentivirus-mediated interleukin-12 gene have an enhanced ability to kill lung cancer cells through promoting T lymphocyte proliferation and cytotoxicity.

• 한국약용작물학회지
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• 제8권4호
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• pp.291-296
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• 2000

구기자나무의 부위별 물추출물이 LPS (lipopolysaccharide) 및 Con A (concanavalin A)에 의한 마우스의 비장세포 증식능에 미치는 영향을 시험한 결과 부위별 면역 활성중 과실에서 의 No mil 처리구에서는 $0.1mg/ml{\sim}0.5mg/ml$ 농도에서, LPS 처리구에서는 $0.01mg/ml{\sim}0.1mg/ml$의 농도에서 B-cell (체액성 면역)의 분열증식을 촉진시켰다. 잎과 뿌리는 LPS 처리구에서 0.1mg/ml 까지 비장세포의 분열증식을 촉진시켰으며 Con A 처리구에서는 T-cell (세포성 면역)의 증식효과가 없었다.

• 대한한방내과학회지
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• 제21권2호
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• pp.243-250
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• 2000

Objectives : Yukmi-Jihwang-Tang(YJT) has been used in Oriental Medicine as a drug for tonifying and nourishing yin. So, the purpose of this Study was to investigate effects of Yukmi-Jihwang-Tang extract(YJTE) on the T-lymphocytes and peritoneal macrophages in mice. Methods : YJTE consists of the following components; Rehmaniae Radix Preparata(熟地黃), Dioscoreae Rhizoma(山藥), Corni-Fructus(山茱萸), Hoelen alba(白茯), Moutan Cortex Radicis(牧丹皮), Alismatis Rhizoma(澤瀉). Result : The results of this Study were obtained as follow; The administration of YJTE did not affect T-lymphocytes apoptosis. YJTE decreased sub-population of TH and TC cells, and proliferation of T-lymphocytes too. But YJTE accelerated phagocytic activity and Nitric Oxide(NO) production from peritoneal macrophages in mice. Conclusions : These results suggest that the administeration of YJTE suppresses T-cell mediated immunity, but activates peritoneal macrophages.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.405-411
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• 2007

Gal-13 is an antimicrobial peptide isolated from chicken intestine. Ninety chickens were randomly divided into two groups (45 chickens for each group) to determine the effect of oral administration of Gal-13 on the acquired immune response. The chickens in the first group were fed a diet without Gal-13 as the control, and the chickens in the second group were fed the same diet, except that Gal-13 ($1{\mu}g/ml$) was suspended in drinking water just after hatching. Samples of blood, thymus, bursa of fabricius and spleen were taken at day 1, 4, 7, 10 and 17. The chickens in both groups received infectious bursal disease virus vaccine at day 20, and then sera samples were collected for analysis at 14, 21, 28 and 35 days after vaccination. The results showed: (1) Gal-13 could enhance the content of immunoglobulin (Ig)G at the age of 4 to10 days (p<0.05) and IgM at the age of 4 and 10 days (p<0.05) in the serum; (2) In vitro experiments showed that Gal-13 (0.625-1.250${\mu}g/ml$) enhanced the proliferation of peripheral blood lymphocytes of the chickens stimulated by lipopolysaccharide (LPS) and concanavlin A (ConA). Compared to the control, Gal-13 (1 ${\mu}g/ml$) enhanced the proliferation of bursa lymphocytes at 17 days of age (p<0.01) and thymus lymphocytes at 7 days of age (p<0.01), but restrained lymphocyte proliferation in chicken spleen and differed significantly at day 10 (p<0.01); (3) Gal-13 enhanced infectious bursal disease virus antibody in sera of chickens 21 days after infectious bursal disease virus vaccine administration (p<0.05). These results suggested that Gal-13 could modulate adaptive immune responses of chickens.