Hongjam is a natural health food that has been shown to have various health-promoting effects, but studies on immunity enhancement have not been done so far. In this study, we investigated whether HongJam extracts could be enhancing innate immunity by protomoting proliferatin of macrophages and their phagocytic or pinocytic abilities to pathogens. (Grant No. PJ017024022023)

Macrophages secrete various cytokines and inflammatory mediators, resulting in playing critical roles in inflammation and immunity. In this study, we investigated anti-inflammatory and immune enhancing properties of PB203, which is a water-soluble extract powder from the fruit of Actinidia polygama, in macrophages. A. polygama is a medicinal plant traditionally known to treat abdominal pain, stroke and rheumatoid arthritis. However, the molecular mechanism for the immune modulation of PB203 is still unclear. Therefore, we assessed the effects of PB203 on the lipopolysaccharide (LPS)-induced inflammation and immune activation, and elucidated its action mechanism in mouse macrophage, RAW264.7 cells. PB203 significantly suppressed not only the levels of nitric oxide (NO), prostaglandin E2, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), but also the mRNA expression of inducible NO synthase, cyclooxygenase-2, TNF-α and IL-1β in LPS-stimulated RAW264.7 cells. We also found that these anti-inflammatory activities of PB203 were mediated through the inhibition of toll-like receptor 4 and nuclear factor kappa B (NF-κB) induced by LPS. On the other hand, in normal macrophages, PB203 dose-dependently elevated the gene expression of immunomodulators including granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor, monocyte chemoattractant protein-1 and TNF-α in a statistically significant manner. The expression of IL-10, IL-1β, IL-6, and interferon-γ were also remarkably upregulated by the treatment of 500 μg/mL PB203. In addition, PB203-mediated production of NO and TNF-α was attenuated by NF-κB inhibition in RAW264.7 cells. Interestingly, PB203 promoted the production of nuclear factor erythroid-2-related factor 2, resulting in the increased level of heme oxygenase-1, which is a representative antioxidant enzyme, in both LPS-stimulated and normal RAW264.7 cells. Taken all together, these results suggest that PB203 may have great potential as the candidate of anti-inflammatory agent for improving inflammatory diseases or immune enhancing agent for preventing infectious diseases. Keywords: Actinidia polygama extract (PB203); macrophages; immunomodulator; nuclear factor kappa B (NF-κB); heme oxygenase-1 (HO-1)

Agar, a heterogeneous polymer of galactose, is the main component of the cell wall of marine red algae. It is well established as a safe, non-digestible carbohydrate in oriental countries. Neoagarooligosaccharides (NAOs) prepared by hydrolyzing agar by microbial β-agarase have been reported to show safety. However, their immunological effects have not been reported yet. Thus, the objective of this study was to investigate immune enhancing effects of neoagarooligosaccharides (NAOs) from marine red algae Gelidium elegans in mice by performing ex vivo experiments. Six-week-old mice were fed ad libitum. NAOs were orally administrated at three different concentrations (100, 500, and 2,500 mg/kg B.W./day) twice a week for four weeks. The group fed with NAOs at 2,500 mg/kg showed the highest proliferation of splenocytes and production levels of cytokines (IL-1β, IL-6, TNF-α) in the ex vivo experiment. In conclusion, NAOs can enhance immune function, increase proliferation of splenocytes, and increase cytokine production by activating macrophages in mice.

The purpose of this study was to investigate probiotic characteristics and immune enhancing effects of Bifidobacterium (B.) longum KBB1-26 and BIF-4, B. breve KBB5-22 isolated from human intestine for probiotic use in humans and animals. We measured acid, bile and heat tolerance, antimicrobial activity against pathogenic bacteria, Escherichia (E.) coli, Salmonella (S.) Enteritidis, Staphylococcus (S.) aureus, and Listeria (L.) monocytogenes. Immune enhancing effects of B. longum and B. breve were investigated by measuring nitric oxide (NO), nuclear factor (NF–κb), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-12 (IL-12) and tumor necrosis factor- α (TNF- α) in RAW 264.7 cells or RAW BLUE cells. B. longum KBB1-26 was survived at pH 2.0. B. longum KBB1-26 and BIF-4, B. breve KBB5-22 also showed tolerance to 0.3% of oxgall bile salt. B. longum KBB1-26 was able to survive at 70 ℃ and 80 ℃ for 20 min. KBB1-26 showed the antimicrobial inhibition zone to pathogenic bacteria such as E. coli (12 mm), S. Enteritidis (14 mm), S. aureus (14 mm) and L. monocytogenes (41 mm). The production of NO (4.5 ± 0.00μM/mL) and IL-1β (39.7 ± 0.55pg/mL) of KBB1-26 significantly higher than BIF-4 and KBB5-22, respectively. In addition, KBB1-26 and KBB5-22 induce the production of high level of TNF-α and IL-6 in macrophages. Collectively, B. longum KBB1-26 have acid, bile, heat tolerance, antimicrobial activity and immune enhancing effects. These results suggest that KBB1-26 can be used as probiotics for humans and animals.

본 연구는 가죽나무(Ailantias altissima) 추출물을 유효성분으로 함유하는 면역 증강용 소재를 개발하기 위하여 수행한 것으로, 가죽나무 에탄올 추출물 단독 또는 면역능을 보유한 유산균과의 혼합처리에 의한 면역 활성능을 비교 분석하였다. 가죽나무 에탄올 추출물에 의한 대식세포 증식에 미치는 영향을 살펴본 결과, 10~500 μg/mL 농도 범위로 처리하였을 때 100 μg/mL 농도에서만 대식세포가 활성화되어 높은 세포 증식률을 나타내었다. NF-κB 활성 평가에서는 유산균과 가죽나무 추출물을 1:0.25 비율로 혼합해서 처리하였을 때 추출물을 단독 처리하였을 때보다 NF-κB 활성능이 증가하였는데, 생균보다 사균과 함께 처리하였을 때 그 값이 더 높았다. 특히 Lactobacillus plantarum 4-25 사균과 추출물을 1:0.25 비율로 혼합 시 가장 높은 활성능을 나타내었다. Nitric oxide(NO) 생성량은 가죽나무 에탄올 추출물의 단독 처리에 의해 5.46±0.06 μM/mL로 LPS 처리 대조군(5.73±0.06μM/mL)과 유사하게 가장 높은 값을 보였지만, 혼합 처리군 에서도 Weissella cibaria JW15와 Lactobacillus rhamnosus GG 등 사균과 가죽나무 추출물을 1:0.25 비율로 처리하였을 때만 대조군보다 높은 생성량을 보였다. TNF-α와 IL-1β 등 사이토카인 생성량에서는 가죽나무 추출물 단독처리군보다 유산균들과의 혼합처리군에서 모두 높은 값을 나타내었는데, 특히 Weissella cibaria JW15 균주를 추출물과 혼합한 경우에만 LPS 처리군보다 높은 TNF-α 활성을 보였고, 생균과 사균 간의 큰 차이 없이 1:0.25 비율로 혼합 시 가장 높은 생성량을 보였다. IL-1β 생성량은 Lactobacillus rhamnosus GG 사균과의 혼합에서는 1:0.25 비율로 처리하였을 때 높은 값을 나타내었고, 가죽나무 추출물 단독 처리와 유사한 수준을 나타내었지만, 나머지 혼합군에서는 IL-1β가 검출되지 않았다. 종합적으로 살펴보면 가죽나무 에탄올 추출물과 유산균주와의 혼합에 의한 면역 활성능을 확인한 결과, 가죽나무 추출물 단독처리보다 유산균과 가죽나무 추출물을 1:0.25 비율로 처리한 혼합물에 의해 높은 면역활성능을 보여, 가죽나무 추출물을 유효성분으로 함유하는 소재를 활용하여 면역 저하증 예방 및 치료용 등 건강기능식품 개발을 위하여 유용하게 이용될 수 있을 것으로 기대된다.

Aging is a growing concern of age-associated aberrations in immune system or immunosenescence. Probiotics contain not only these general health advantages but also other the regulation of host immune function. Among those probiotics, the immunomodulatory effects of Weissella cibaria JW15 (JW15) strain has already been reported in our previous studies. The objective of this study is to assess improved immunity of JW15 strain in aged mice. In experiment, mice were divided into five groups. Twenty eight–month-old C57BL/6J mice were given a daily dose of 1×109 CFU/ mouse (JW15-H group) and 1×108 CFU/mouse (JW15- L group) of viable JW15. The young mice group (YM) and old mice group (OM) were given PBS. After four weeks of administration, mice were euthanized. Mice blood was collected and analyzed for complete blood cell count (CBC). Then, mice spleen was weighed and analyzed for splenocyte. Finally, we measured the concentration of cytokine secreted by splenocyte and serum. The results showed splenocyte proliferation of JW15-L group by the addition of LPS and concanavalin A was significantly higher than that of OM group. Splenocyte TNF-α production was significantly increased by JW15 intake. In particular, splenocyte TNF-α production of JW15-L group by the addition of LPS (5 μg/mL) was significantly greater than that other group. Splenocyte IL-6 production was also the highest in JW15-L group. Serum IFN-γ production of JW15-L (59.13 ± 16.13 pg/mL) group was significantly higher than that of OM group (39.71 ± 2.51 pg/ mL). When compared to the OM group (32.22 ± 1.92 pg/mL), Serum TNF-α production was significantly increased by JW15-H intake (69.01± 26.51 pg/mL). In conclusion, JW15 enhanced immunomodulative effects. In particular, the effect was significantly excellent in JW15-L group (1 × 108 CFU/mouse). Therefore, Weissella cibaria JW15 would be suitable for consideration as a helpful functional food for companion animals and humans.

Lactic acid bacteria (LAB) are known to promote activation of macrophages, selectively eliminate harmful intestinal bacteria that enter the body, and increase secretion of cytokines, chemokines, and inflammatory mediators that boost the immune system. In the current study, the immune-enhancing effects of five Leuconostoc stains (Leu. kimchii WK18, Leu. citreum DH33, Leu. mesenteroides DH34, SE34, and GY17) isolated from traditional Korean fermented food, Kimchi, were evaluated. Lactobacillus rhamnosus GG (LGG), which is known for its extraordinary immunopotentiating activity, was used as a reference strain. According to the results, several heat-killed Leuconostoc strains induced a higher level of NO, IL-1β, or TNF-α production than the LGG strain. Collectively, results of the current study suggest that Leuconostoc strains could enhance immune response through activation of macrophages, and could be significant references when verifying the possibility for use of Leuconostoc strains as probiotics, starter, dietary supplement, or immune-enhancing medicine.

Recently, as a natural substance has been emphasized interest in research to enhance the immune function. Green lettuce (Lactuca sativa L.) is a popular vegetable used fresh and it contains various phytochemicals and antioxidant compounds, and has been reported to have various physiological activities such as antibacterial, antioxidant, antitumor and anti-mutagenic. However, only a few studies have investigated on the mechanism of action of immune-enhancing activity of lettuce. Therefore, in this study, the immunomodulatory activities and potential mechanism of action of Green lettuce extracts (GLE) were evaluated in the murine macrophage cell line RAW264.7. GLE significantly increased NO levels by RAW264.7 cells, as well as expressions of immunomodulators such as iNOS, COX-2, IL-1β, IL-6, IL-12, TNF-α and MCP-1. Although GLE activated ERK1/2, p38, JNK and NF-κB, GLE-mediated expressions of immunomodulators was dependent on p38, JNK and NF-κB. In addition, TLR4 inhibition blocked GLE-mediated expressions of immunomodulators and activation of p38, JNK and NF-κB. Taken together, these results demonstrated that TLR4-MAPK/NF-κB signalling pathways participated in GLE-induced macrophage activation and GLE could be developed as a potential immunomodulating functional food.

This study investigated the immune-enhancing effects of polysaccharides extracted from Gloiopeltis furcata (red seaweed) with different molecular weights. A crude polysaccharide mixture was hydrolyzed using acid treatment (0.1 N HCl) and three molecular weight fractions were generated and filtered using centrifugation: (≤10 kDa, 10 to 100 kDa, and 100 kDa. Nitric oxide (NO) production in RAW264.7 cells treated with 0.01-0.5 μg/mL polysaccharides ≥100 kDa was 12.28-19.05 μM. Treatment with polysaccharides ≥100 kDa increased cytokine levels, including TNF-α and IL-6 levels, in a dose-dependent manner. Polymerase chain reaction analysis also revealed marked increases in iNOS and COX-2 mRNA expression levels. These findings lead us to conclude that macrophage activation induced by polysaccharides ≥100 kDa was greater than that induced by polysaccharides ≤10 kDa or between 10 and 100 kDa. The polysaccharides ≥100 kDa extracted from Gloiopeltis furcata investigated herein are potentially useful as natural immune-enhancing agents. These findings provided further insights into the potential use of ≥100 kDa as immunopotentiator or new function food.

국화과 약용식물들의 각 추출물은 정상세포에 대한 생육도를 측정한 결과, 두 종류 모두 0.5g/L 이하의 농도로 투여시는 정상세포 생존율을 85%이상으로 유지시켜 정상세포에 대한 안정성이 유지되었다. 또한 암세포 생육억제활성의 경우, 국화과 Herb들은 증류수 및 에탄올 추출물 모두 0.5mg/ml이상의 농도에서 유방암세포주인 MCF7에 대하여 70%이상, 폐암세포주인 A549에 대하여 60%이상의 높은 활성을 나타내었다. 간암세포주인 Hep3B에 대하여서는 증류수 추출물들은 0.5mg/ml이상의 농도에서 약 40%정도, 에탄올 추출물들은 70%이상의 높은 암세포 생육억제활성을 나타내었으며, 위암세포주인 AGS에 대하여 증류수 추출물들은 0.5mg/ml이상의 농도에서 약 60%정도, 에탄올 추출물들은 80%이상의 높은 암세포 생육억제활성을 나타내었다. 정상세포에 대한 암세포의 상대적 사멸도인 selectivity에 있어서 각 추출물이 0.1mg/ml에서 1.0mg/ml의 농도에서 3이상의 수치를 나타내어 암세포를 선택적으로 사멸시키는 기작을 나타내었다. 또한 인간 B 세포와 T 세포에 대한 생육촉진 실험 결과, Echinacea, 엉겅퀴의 증류수 추출물과 Echinacea 에탄을 추출물은 0.5 mg/ml 이상의 농도에서 T-cell의 생장을 2배 이상, B-cell의 경우 1.5배 이상의 생육을 증진시켰으며, Echinacea 경우 증류수 및 에탄올 추출물에서, 0.5mg/ml이상의 농도에서 T-cell의 생장을 2~2.6배 이상 B-cell의 경우 1.4배 이상의 생육증진효과를 나타내었다. 시료들의 IL-6와 TNF-α의 cytokine생성량을 측정한 결과 IL-6는 엉겅퀴의 에탄올 추출이 0.5mg/ml의 농도에서 96pg/ml의 분비량을 TNF-α는 Echinacea의 에탄을 추출물이 94pg/ml의 분비량을 나타내었다. Echinacea의 에탄올 추출물의 인간 간암 세포에 대한 산화는 추출물 투여 후 세포의 산화가 급격히 나타났다. 위의 연구들을 미루어 볼 때 국화과 약용식물들은 기능성 식품 소재로서의 가능성이 매우 높은 것으로 사료되어진다.