• Biomolecules & Therapeutics
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• 제2권3호
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• pp.236-243
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• 1994

A splenocyte culture system supplemented with liver microsomes was developed to detect immunotoxic chemicals which require metabolic activation using cyclophosphamide as a positive standard. When liver microsomes were added to splenocyte cultures isolated from female B6C3Fl mice, the proliferation of splenocytes by lipopolysaccharide (LPS) was increased and the proliferation by concanavalin A (Con A) was decreased. However, when compared with each corresponding control, cyclophophamide was successfully activated to metabolites capable of suppressing Iymphoproliferative responses. This suppression was clearly dependent upon the amounts of microsomes added and/or the concentration of cyclophosphamide exposed. In these cultures, the proliferation of splenocytes was suppressed when the cells were exposed to cyclophosphamide on the day of culture initiation. On the other hand, microsome was responsible for the increase in LPS mitogenicity and NADPH was responsible for the decrease in Con A mitogenicity. Finally, our present culture system was compared with the hepatocyte-splenocyte coculture system which we had developed earlier. We found that the hepatocyte-splenocyte coculture was better able to activate cyclophosphamide to metabolites capable of suppressing the antibody response to sheep erythrocytes. Although our present culture system was relatively poor to activate cyclophosphamide in cultures for antibody response, it will be useful as a simple screening method to detect suppression of certain in vitro immunotoxic parameters like LPS mitogenicity by chemicals which require metabolism.

• 약학회지
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• 제45권1호
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• pp.55-63
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• 2001

In order to investigate the effects of bisphenol A (BPA) on immune system in mice we examined the various immunological parameters. After single oral administration of BPA to female ICR mice, the weights of bodies and lymphoid organs, splenic cellularity and hematological parameters were examined on day 2 and 7. Among them WBC and splenic cellularity were slightly decreased on day 2. To assess the effects of BPA on humoral immune responses, splenic IgM plaque forming cell (PFC) and serum IgM were assayed. When BPA was administered after immunization with SRBC, but not before immunization, IgM PFC against SRBC was significantly lowered in a dose dependent manner. Serum IgM level was also decreased on day 4 when high dose (2000 mg/kg) of BPA was administrated after injection of OVA-antigen. The indexes of splenocyte proliferation (SP) to concanavalin A (Con A) and bacterial lipopolysaccharide (LPS) were measured in vitro by MTT assay. At low concentration BPA slightly increased splenocyte proliferation but at higher concentration it showed significant inhibitory effects on cell proliferation. Mitogen-stimulated SP was also determined with spleen cells from BPA treated mice. Con A-induced SP was slightly decreased and LPS-induced SP was especially inhibited at 1000 mg/kg and 2000 mg/kg of BPA. These results indicate that BPA is able to acutly evoke humoral and cell mediated immune suppression in mice.

• Nutrition Research and Practice
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• 제17권2호
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• pp.206-217
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• 2023

BACKGROUND/OBJECTIVES: The immunomodulatory effect of Platycodon grandiflorum (PG) has been reported, but studies on its mechanism are still lacking. This study was undertaken to confirm whether the hydrolyzed and fermented PG extract (HFPGE) obtained by adding hydrolysis and fermentation to the extraction process has an immune-enhancing effect in the in vivo system. MATERIALS/METHODS: Five-week-old BALB/c mice were divided into 4 groups: normal control group (NOR), control group (CON), 150 mg/kg body weight (BW)/day HFPGE-treated group (T150), and 300 mg/kg BW/day HFPGE-treated group (T300). The mice were administered HFPGE for 4 weeks and intraperitoneally injected with cyclophosphamide (CPA, 80 mg/kg BW/day) on day 6, 7, and 8, respectively, to induce immunosuppression. The levels of immunoglobulins (Igs) and cytokines were measured in the serum. In splenocytes, proliferation and cytokine levels were measured. RESULTS: Serum IgA, IgG, and IgM levels were observed to decrease after CPA treatment, which was recovered by HFPGE administration. The levels of serum interleukin (IL)-12, tumor necrosis factor (TNF)-α, IL-8, and transforming growth factor (TGF)-β were also decreased after exposure to CPA but increased after HFPGE administration. Decreased splenocyte proliferation was seen in CPA-treated mice, but was observed to increase in the T150 and T300 groups as compared to the NOR group. Compared to the CON group, splenocyte proliferation stimulated with concanavalin A (ConA) or lipopolysaccharide (LPS) in the HFPGE-treated groups was significantly increased. The cytokines secreted by ConA-stimulated splenocytes (IL-2, IL-12, interferon-γ, TNF-α) were increased in the T150 and T300 groups, and cytokines secreted by LPS-stimulated splenocytes (IL-4, IL-8, TGF-β) were also increased by HFPGE administration. CONCLUSION: These results suggest that HFPGE stimulates the immunity in immunosuppressed conditions, thereby enhancing the immune response. Therefore, it is expected that HFPGE has the potential to be used as functional food and medicine for immune recovery in various immunocompromised situations.

• 대한미생물학회지
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• 제21권1호
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• pp.163-170
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• 1986

Balb/c mouse spleen cells in vitro sensitized against ICR spleen cells were cultured in conditioned media(CM). The CM was produced by ICR spleen cells stimulated with Concanavalin-A(Con-A), and sensitized lymphoid cells were grown in CM. ICR mouse spleen cells were appeared to be a good generator of IL-2. Optimal growth was seen in growth medium containing 20% fetal calf serum. and 25% CM. When cultures were initiated at 1, 5, $10{\times}10^4\;cells/ml$, the cells were increased in numbers by about 20, 13, 5-fold, respectively, every 9 days. Such growth pattern was sustained for about 4-6 weeks and thereafter the cell growth was diminished gradually. Direct immunofluorescence indicated that 93% of the lymphoid cells grown in CM(for 10 days) expressed Thyl surface antigen. And the cells grown in CM were cytotoxic to the sesitizing ICR mouse spleen cells though cytotoxicity level was not high. According to these results, the cells grown in CM were considered to be cytotoxic T lymphocytes. The lymphoid cells grown for 20 days were nearly unresponsive to Con-A and therefore dependent only IL-2 to be used for IL-2 assay.

• 약학회지
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• 제50권3호
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• pp.184-190
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• 2006

We investigated the effect of fresh Rehmanniae radix methanol extracts (RGMeOH) on the in vitro production of cytokines by splenocytes and peritoneal macrophages isolated from C57BL/6 mice. Peritoneal macrophages and splenocytes were incubated with various concentrations of RGMeOH in the presence of $10\;{\mu}g/ml$ of lipopolysaccharide (LPS) $1\;{\mu}g/ml$ of concanavalin A (Con A) for cytokine assay, These results showed that RGMeOH remarkably attenuated LPS-increased production of $TNF-{\alpha}$ but not IL-6 by peritoneal macrophages and enhanced LPS-stimulated production of IL-10 in a dose-dependent manner RGMeOH significantly augmented the LPS- or Con A-stimulated production of IL-2 and $IFN-{\gamma}$ by splenocytes. These findings suggest that RGMeOH may attenuate inflammatory responses through down-regulation of $TNF-{\alpha}$ and up-regulation of IL-10, and that RGMeOH may up-regulate cell-mediated immune responses through increase in IL-2 and $IFN-{\gamma}$ production.

• 동의생리병리학회지
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• 제19권4호
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• pp.908-912
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• 2005

Bio-Q is a modified prescription with the activities of supplementing Qi and blood in human body. In the present study, immunomodulatory effect of Bio-Q was examined. After oral administration of Bio-Q for 7 days to Balb/c mice, splenocytes were isolated and immunological experiments were performed. Bio-Q significantly increased the proliferation of splenocytes exposed to concanavalin A (Con A), while it did not in case of lipopolysaccharide (LPS) stimulation. Bio-Q also significantly increased CD3/CD19, CD4/CDB and NK cells by flow cytometric analysis. In addition, Bio-Q significantly enhanced the level of $INF-\gamma$ in splenocytes, but not $TNF-\alpha$ by ELISA. These results strongly suggest that Bio-Q has immunomodulatory activity through the regulation of T cell mediated immune pathway.

• 사상체질의학회지
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• 제25권3호
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• pp.218-232
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• 2013

Objectives The purpose of this study was to investigate the effect of Yongyukjowitang(YJT) distillate on the immune activity of spleen cells of aged SD rats. Methods Spleen cells from 10w, 52w, 72w old rats were stimulated with Concanavalin A(Con A) and treated with Vitamin C(Vit.C) or Yongyukjowitang distillate(YJT). After 24 hours, levels of IL-2, IL-4, IL-10, IFN-${\gamma}$ were measured using ELISA in spleen cells. Results and Conclusions 1. The concentration of IL-2, IL-4 in spleen cells of 52 weeks old SD rats(YJT group) significantly decreased in comparison with ConA group. 2. The concentration of IFN-${\gamma}$ in spleen cells of 72 weeks old SD rats significantly increased in comparison with 10 weeks old SD rats(NT group), but decreased in comparison with 10 weeks old SD rats(YJT group). These results suggest that Yongyukjowitang distillate has an effect on the immune activity of spleen cells of aged SD rats.

• 한국어병학회지
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• 제20권2호
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• pp.189-200
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• 2007

잉어의 선천성 면역 인자가 관여하는 항바이러스 면역 반응을 조사하기 위해 UV-inactivated SVCV, Poly I:C 및 Con A를 주사한 후 3일째 라이소자임 활성, 혈청 내 보체의 살균능력 및 식세포의 활성산소량을 조사하였다. 그 결과, 모든 시험구에서 혈청 내 라이소자임의 활성은 유의적인 차이를 나타나지 않았으나, 두신 조직의 라이소자임 활성은 대조구에 비해 유의적으로 증가하였다. 또한, 혈청 내 보체의 살균 능력도 모든 시험구에서 대조구와 유의적인 차이가 없었다. 그러나 식세포의 활성은 UV-inactivated SVCV 시험구에서는 농도에 따라 증가한 것으로 나타났으며, Poly I:C 및 Con A 시험구에서는 저농도에서 활성이 증가한 것으로 나타났다. 바이러스에 대한 방어력을 조사하기 위해 주사 후 4일째 1×104 TCID50/fish 농도의 SVCV로 인위 감염한 결과 UV-inactivated SVCV 및 Poly I:C 시험구에서는 Con A 시험구에 비해 높은 방어력을 나타내었다. 또한, Poly I:C 시험구에서 라이소자임 및 식세포 활성이 다소 감소한 고농도에서도 높은 방어력이 유도된 것으로 나타나 이러한 결과는 Poly I:C에 의해 자극된 또 다른 비특이적 면역 인자가 SVCV에 대한 방어반응에 관여한 것으로 추정된다.

• Journal of Ginseng Research
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• 제32권3호
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• pp.264-269
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• 2008

Ginsenosides are major components in Panax ginseng and known to have numerous pharmacological activities such as anti-cancer, anti-diabetes, anti-viral and anti-atherosclerosis effects. In this study, the regulatory activities of G-Rg3 and its derivative 25-hydroxy Rg3 (G-Rg3-2H) on the production of nitric oxide (NO) in macrophages and the proliferation of lymphocytes prepared from spleen and bone marrow under treatment of lipopolysaccharide (LPS) or concanavalin (Con) A were examined. G-Rg3 and G-Rg3-2H dose-dependently inhibited NO production from LPS-activated RAW264.7 cells and in agreement, these compounds protected RAW264.7 cells from LPS-mediated cytotoxicity. In contrast, G-Rg3-2H dose-dependently inhibited lymphocyte proliferation induced by both LPS and Con A, while there was no inhibition by G-Rg3. Therefore, our data suggest that these compounds may be applied for NO-mediated or lymphocyte-mediated immunological diseases.

• The Korean Journal of Physiology and Pharmacology
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• 제1권1호
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• pp.91-96
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• 1997

Lipocortin 1 has been proposed as a putative mediator of anti-inflammatory actions of glucocorticoids. We investigated the role of lipocortin 1 in the effect of dexamethasone using rat splenic leukocytes. Concanavalin A(ConA; 1 ${\mu}g/ml$) increased the leukocyte proliferation and nitric oxide(NO) generation, which were measured as $[^3H]-thymidine$ uptake by the cells and nitrite accumulation in the culture media, respectively. Dexamethasone suppressed ConA-induced cell proliferation, in a concentration-dependent manner with $EC_{50}$ around 50nM. The addition of anti-lipocortin l(Anti-LCl) reversed dexamethasone effects: 0.24, 1.2, 6 ${\mu}g/ml$ of Anti-LC1 reversed dexamethasone(50 nM)-induced suppression of thymidine uptake by $9{\pm}3%$, $16{\pm}3%$, $36{\pm}5%$, respectively; 0.24, 1.2, and 6 ${\mu}g/ml$ of Anti-LCI reversed dexa-methasone-induced decrease of nitrite concentration by $49{\pm}16%$, $61{\pm}20%$, $77{\pm}19%$, respectively. The present data indicate that lipocortin 1 mediates, at least in part, glucocorticoids-induced suppression of leukocyte proliferation and blockade of NO generation.