Nelumbo nucifera Gaertn has been usedas a traditional remedy and food source in South Korea. It promotes gastrointestinal function and controls blood pressures. Nelumbo nucifera Gaertn water extracts supplement at 5, 10, 50, 100, 250, 500, 1,000 μg/mL after a 48 h pre-treatment with the mitogen (ConA or LPS) increased the mouse splenocytes proliferation. Water extract supplement also increased the cytokine production (IL-1β, TNF-α and IFN-γ), measured by a cytokine ELISA kit. For the result of in vitro study, the proliferation of splenocytes and cytokine production activated by peritoneal macrophages increased when water extracts were supplemented in the range of 50~500 μL/mL concentration. Specifically, the levels of the splenocytes proliferation, IL-1β, TNF-α and IFN-γ were the highest at 250 μL/mL concentration. This in vitro study suggestedthat supplementation with Nelumbo nucifera Gaertn water extracts may enhance the immune function by regulating the splenocyte proliferation and enhancing the cytokine production activating macrophage in vitro.

Disulfiram is a drug used to treat alcohol dependence. Recent studies have shown that disulfiram also has anti-cancer effects. Considering that many anti-cancer agents have side effects, including immunosuppression, it is important to check if disulfiram has some cytotoxicity to immune cells. In this study, mouse spleen cells were treated with disulfiram and the metabolic activity was measured. Disulfiram increased the cell death of spleen cells according to annexin V-FITC/PI staining analysis. In addition, disulfiram decreased the mitochondrial membrane potential of spleen cells. The toxicity of disulfiram was concentration dependent. Interestingly, disulfiram affected the population of lymphocytes and the subset of spleen cells was altered. This study provides clinicians and researchers with valuable information regarding the toxicity of disulfiram to mouse spleen cells, particularly lymphocytes.

Doxorubicin has been used to treating cancers, including breast cancer, bladder cancer, and acute lymphocytic leukemia, however, few studies have investigated its anti-inflammatory activity. In this study, we used mouse spleen cells treated with lipopolysaccharide (LPS), a representative inflammatory agent to investigate the effects of doxorubicin. Specially, we investigated the effects of doxorubicin on metabolic activity, cell size, cell death, and cytokine production of LPS-treated spleen cells. Doxorubicin significantly decreased the metabolic activity, even when applied at relatively low concentrations (1.6-8 ng/mL). To investigate the potential mechanism, we measured the mitochondrial membrane potential (MMP) of the LPS-treated spleen cells using Rhodamine 123. Doxorubicin decreased MMP and cell size, and induced cell death. Furthermore, doxorubicin suppressed the production of tumor necrosis factor (TNF)-alpha, a representative cytokine, in LPS-treated spleen cells. Taken together, doxorubicin decreased metabolic activity and the production of inflammatory cytokines, while increasing the death of LPS-induced hyperactivated spleen cells. This results will enable broader application of doxorubicin, as an anti-inflammatory agent, in clinical and research fields.

King oyster mushroom (Pleurotus eryngii), an improved species of oyster mushroom, is a popular ingredient in Asian cuisine. Spleen cells were treated with various concentrations (0, 5, 10, 50, 100, 250, 500, and 1,000 μg/mL) of king oyster water extracts (KOWE); then, the proliferation of the cells was measured 24, 48, and 72 h after each treatment. Also, type 1 T helper cytokine productions (TNF-α, IFN-γ, and IL-2) were measured in activated macrophage by KOWE in seven concentrations. Under the condition of its 50, 100, 250, and 1,000 μg/mL for 48 h, the proliferation of cells was increased. However, there was no significant fluctuation in the spleen cells proliferation for 24 and 72 h-long KOWE exposure. To determine cytokine (TNF-α, IFN-γ, IL-2) productions of type 1 T helper cells, macrophage was stimulated by KOWE for 48 h. Treatment of KOWE gave a rise to the levels of TNF-α and IFN-γ, but not in that of IL-2 productions. These results suggest that king oyster mushroom water extracts may be beneficial for enhancing immune functions in its high concentration.

Plantago asiatica L., observed frequently in East Asia, is a known herb used in traditional medical remedies several studies report that P. asiatica L has anti-inflammatory and antioxidant effects. To determine the production of cytokines (IL-2, IFN-γ, and TNF-α) induced by lipopolysaccharide (LPS) and non-LPS-stimulated macrophages, an ELISA assay was conducted using cytokine kits. Mice splenocytes were cultured for 48 h with various concentrations of P. asiatica L. (5, 10, 50, 100, 250, 500, and 1,000 μg/mL) or with mitogens (ConA or LPS). P. asiatica L. increased the proliferation of mice splenocytes, especially under the condition of its concentration ranging from 250 to 1,000 μg/mL. In addition, Plantago asiatica L. notably induced cytokine production of (IL-2, IFN-γ, and TNF-α) at its concentration of 250~500 μg/mL. These results suggest that supplementation with P. asiatica L. water extracts may play a potential role in enhancing immune function by mediating splenocyte proliferation and cytokine production through its anti-inflammatory activit.

Fucoidan is a sulfated polysaccharide, purified from brown algae. It has multiple biological activities including anti-cancer and anti-inflammatory effects. Our previous reports demonstrated that fucoidan can stimulate spleen cells and especially high molecular weight fucoidan is responsible for the immunostimulatory activity. However, we recently found that the activity of fucoidan can be dependent on its individual batch or sources. Four different fucoidans (fucoidan A, fucoidan B, high molecular weight fucoidan, and low molecular weight fucoidan) were used for this study. MTT assay and flow cytometry analysis were performed for analysis of the activity of fucoidan. MTT assay showed that fucoidan B significantly decreased the cellular activity of spleen cells compared to fucoidan A. In addition, fucoidan B consistently killed spleen cells based on the cell size by flow cytometry analysis and the morphology by an inverted microscope. To elucidate the detailed mechanisms of cytotoxicity, fucoidan B-treated spleen cells were stained with Rhodamine 123 solution and Annexin V-FITC/propidium iodide for measurement of mitochondrial membrane potential (MMP) and early/late apoptosis, respectively. From these two assays, fucoidan B decreased the MMP and induced early apoptosis of spleen cells. Taken together, we suggest that different batches or origin of fucoidan may have differential activities on spleen cells, immunostimulatory and cytotoxic activity. The present study may provide some valuable information regarding use of fucoidan in the clinical area and in basic research.

Taxol is an anti-cancer agent that stabilizes the microtubules of cancer cells, resulting in inhibition of mitosis and thus preventing the proliferation of cancer cells. However, many anti-cancer agents including taxol work on normal cells as well as cancer cells, resulting in side effects such as immunosuppression. A marine algae-derived sulfated polysaccharide, fucoidan, an anti-cancer agent, also showed immunostimulating effects. This study investigated the effects of fucoidan on taxol-treated spleen cells. Spleen cells were treated with taxol in a concentration-dependent manner and in combination with fucoidan. MTT assay and flow cytometry analysis were performed to measure the viability and activity of treated cells. Two assays demonstrated that taxol induced the death of spleen cells. Fucoidan clearly inhibited the cell death induced by taxol. In addition, fucoidan enhanced the production of nitric oxide in spleen cells, which was decreased by taxol. Taken together, taxol can induce the cell death of spleen cells, a major type of immune cells and fucoidan protects spleen cells from taxol-induced cell death. This finding suggests that taxol and fucoidan can be used in combination for lowering the immunosuppressive effects of taxol.

In vitro 실험을 통한 가시오가피 물 추출물 첨가가 마우스 의 면역세포 증식에 미치는 영향에 대한 연구 결과, 음의 대 조군에 비해 가시오가피 물 추출물을 첨가한 모든 농도에서 비장세포 증식능이 증가하였으며, 특히 고농도인 500~1,000 μg/ mL 농도에서 유의적으로 증가하였다. 반면, 복강 대식세 포로부터 유도된 사이토카인 생성의 경우, IL-2, IFN-γ, TNF- α 사이토카인 생성량을 측정한 결과, TNF-α 사이토카인은 가시오가피 물 추출물 50 μg/mL 농도와 250~1,000 μg/mL 농 도에서 대조군보다 유의적으로 높은 분비량을 보인 반면, IFN-γ에서는 변화를 보이지 않았다. IL-2의 경우, 100~500 μg/ mL에서 유의적으로 증가하는 경향을 보여주었다. 이상의 결 과에 의하면 가시오가피 물 추출물은 마우스 비장 세포를 증 식시키고, 사이토카인 분비량에도 영향을 줄 것으로 보이며, 면역 기관의 주요 기능을 증진시킬 가능성이 있을 것으로 사료된다. 따라서 가시오가피 물 추출물이 면역 증진 기능성식 품 개발의 소재로 활용될 가능성이 있을 것으로 기대된다

In vitro 실험을 통한 느타리버섯 물 추출물 첨가가 마우스의 면역세포 증식에 미치는 영향에 대한 연구 결과, 음의 대조군에 비해 느타리 물 추출물을 첨가한 모든 농도에서 비장세포 증식능이 증가하였으며, 특히 100~1,000 μg/mL 농도에서 유의적으로 증가하였다. 반면, 사이토카인 생성의 경우, IL-2, IFN-γ, TNF-α 사이토카인 생성량을 측정한 결과, 느타리버섯 물 추출물 50~500 μg/mL 농도에서 생성된 IL-2, IFN-γ, TNF-α 사이토카인은 대조군보다 높은 생성량을 보였다. IFN-γ, TNF-α 사이토카인 모두 50, 100, 250, 500 μg/mL의 농도에서 높은 생성량을 나타내었다. IL-2의 경우, 생성량이 유의적인 차이는 보이지 않았지만, 50 μg/mL 이상에서 증가하는 경향을 보여주었다. 이상의 결과에 의하면 느타리버섯 물 추출물은 마우스 비장 세포를 증식시키고, 사이토카인 생성량에도 영향을 주어, 면역 기관의 주요 기능을 증진시킬 가능성이 있을 것으로 사료되며, 느타리버섯이 면역 증진 기능성 식품 개발의 소재로 활용될 가능성이 있을 것으로 기대된다.

Mycoplasma hyopneumoniae (M. hyopneumoniae) is one of the causative bacteria that can induce chronic enzootic pneumonia, resulting in low production in the swine industry. Potentiation of porcine reproductive and respiratory syndrome virus-induced pneumonia by M. hyopneumoniae has also been recognized. Although some available vaccines have been developed for prevention of M. hyopneumoniae infection, protective immunity is still poor. In this study, in order to provide valuable information on vaccine antigen, we investigated the immunogenicity of M. hyopneumoniae on mouse spleen cells. Concanavalin A (ConA) and lipopolysaccharide (LPS) were used for generation of activated T and B lymphocytes. M. hyopneumoniae made clusters of spleen cells and also affected the cellular activity and viability of spleen cells by alone or with mitogens. Of particular interest, it induced a significant increase in production of TNF-alpha in ConA- treated spleen cells, meaning T helper 1 response. In addition, cell size and mitochondrial membrane potential of M. hyopneumoniae–treated spleen cells were measured by flow cytometric analysis. M. hyopneumoniae did not affect the cell size by alone, whereas ConA or LPS profoundly increased the cell size. Taken together, M. hyopneumoniae significantly affect the cellular activity and cytokine production of spleen cells by alone or in a combination of ConA. This study provides valuable information for production of the vaccine against M. hyopneumoniae.

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~0.5mg/ml 농도에서, LPS 처리구에서는 0.01mg/ml~0.1mg/ml의 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.