• Title/Summary/Keyword: Immune Cells

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Activation of the NF-$\kappa$B p50/p65 Complex in Human Lung Cancer Cell Lines (인체 폐암세포주에서 NF-$\kappa$B p50/p65 Complex의 활성화)

  • Choi, Hyung-Seok;Yoo, Chul-Gyu;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.2
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    • pp.185-194
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    • 1999
  • Background: NF-$\kappa$B is a characteristic transcriptional factor whose functional activity is determined by post-translational modification of protein and subsequent change of subcellular localization. The involvement of the NF-$\kappa$B family of the transcription factors in the control of such vital cellular functions as immune response, acute phase reaction, replication of certain viruses and development and differentiation of cells has been clearly documented in many previous studies. Several recent observations have suggested that the NF-$\kappa$B might also be involved in the carcinogenesis of some hematological and solid tumors. Investigating the possibility that members of the NF-$\kappa$B family participate in the molecular control of malignant cell transformation could provide invaluable information on both molecular pathogenesis and cancer-related gene therapy. Method: To determine the expression patterns and functional roles of NF-$\kappa$B family transcription factors in human lung cancer cell lines NCI-H792, NCI-H709, NCI-H226 and NCI-H157 were analysed by western blot, using their respective antibodies. The nuclear and the cytoplasmic fraction of protein extract of these cell lines were subsequently obtained and NF-$\kappa$B expression in each fraction was again determined by western blot analysis. The type of NF-$\kappa$B complex present in the cells was determined by immunoprecipitation. To detect the binding ability of cell-line nuclear extracts to the KB consensus oligonucleotide, electrophoretic mobility shift assay(EMSA) was performed. Results: In the cultured human lung cancer cell lines tested, transcription factors of the NF-$\kappa$B family, namely the p50 and p65 subunit were expressed and localized in the nuclear fraction of the cellular extract by western blot analysis and immunocytochemistry. Immunoprecipitation assay showed that in the cell, the p50 and p65 subunits made NF-$\kappa$B complex. Finally it was shown by Electrophoretic Mobility Shift Assay(EMSA) that nuclear extracts of lung cancer cell lines are able to bind to NF-$\kappa$B consensus DNA sequences. Conclusion: These data suggest that in human lung cancer cell lines the NF-$\kappa$B p50/p65 complex might be activated. and strengthen the hypothesis that NF-$\kappa$B family transcription factors might be involved in the carcinogenesis of human lung cancer.

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IL-12 Regulates B7-H1 Expression in Ovarian Cancer-associated Macrophages by Effects on NF-κB Signalling

  • Xiong, Hai-Yu;Ma, Ting-Ting;Wu, Bi-Tao;Lin, Yan;Tu, Zhi-Guang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5767-5772
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    • 2014
  • Background and Aim: B7-H1, a co-inhibitory molecule of the B7 family, is found aberrantly expressed in ovarian cancer cells and infiltrating macrophage/dendritic-like cells, and plays a critical role in immune evasion by ovarian cancer. IL-12, an inducer of Th1 cell development, exerts immunomodulatory effects on ovarian cancer. However, whether IL-12 regulates B7-H1 expression in human ovarian cancer associated-macrophages has not been clarified. Therefore, we investigated the effects of IL-12 on the expression of B7-H1 in ovarian cancer-associated macrophages and possible mechanisms. Methods: PMA induced THP-1-derived macrophages or human monocyte-derived macrophages were treated with recombinant IL-12 (rIL-12) or infected with adenovirus carrying human IL-12 gene (Ad-IL-12-GFP) for 24 h, then cocultured with the SKOV3 ovarian cancer cell line for another 24 h. Macrophages were collected for real-time PCR and Western blot to detect the expression of B7-H1, and activation of the NF-${\kappa}B$ signaling pathway. Moreover, supernatants were collected to assay for IL-12, IFN-${\gamma}$ and IL-10 by ELISA. In addition, monocyte-derived macrophages treated with IFN-${\gamma}$ were cocultured with SKOV3 and determined for the expression of B7-H1. Furthermore, the expression of B7-H1 in monocyte-derived macrophages was also evaluated after blocking NF-${\kappa}B$ signaling. Results: The expression of B7-H1 was significantly upregulated in monocyte-derived macrophages treated with rIL-12 or Ad-IL-12-GFP compared with the control groups (p<0.05), accompanied by a remarkable upregulation of IFN-${\gamma}$ (p<0.05), a marked downregulation of IL-10 (p<0.05) and activation of NF-${\kappa}B$ signaling. However, the upregulation of B7-H1 was inhibited by blocking the NF-${\kappa}B$ signaling pathway (p<0.05). Expression of B7-H1 was also increased (p<0.05) in monocyte-derived macrophages treated with IFN-${\gamma}$ and cocultured with SKOV3. By contrast, the expression of B7-H1 in THP-1-derived macrophages was significantly decreased when treated in the same way as monocyte-derived macrophages (p<0.05), and IL-10 was also significantly decreased but IFN-${\gamma}$ was almost absent. Conclusions: IL-12 upregulates the expression of B7-H1 in monocyte-derived macrophages, which is possible though inducing the secretion of IFN-${\gamma}$ and further activating the NF-${\kappa}B$ signal pathway. However, IL-12 downregulates the expression of B7-H1 in THP-1-derived macrophages, associated with a lack of IFN-${\gamma}$ and inhibition of expression of IL-10.

Effect of Surfactin on Growth Performance of Weaning Piglets in Combination with Bacillus subtilis BC1212 (바실러스 섭틸리스 BC1212와 설팩틴의 병용투여가 이유돈의 성장에 미치는 영향)

  • Kim, Myoung-Seok;Lim, Jong-Hwan;Park, Byung-Kwon;Hwang, Yun-Hwan;Song, In-Bae;Park, Seung-Chun;Yun, Hyo-In
    • Journal of Veterinary Clinics
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    • v.26 no.2
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    • pp.117-122
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    • 2009
  • The aim of this study is to investigate the effects of surfactin in combination with Bacillus subtilis BC1212 isolated from Korean soybean paste, on feed utilization and growth performance during 4 weeks in weaning piglets. Eighteen weaning piglets(Landrace$\times$Yorkshire$\times$Duroc; weighing $7.68{\pm}0.97\;kg$) were divided into control(n=9) and experimental groups(n=9). The treatments included a control group consisting of the basal diet with no additives(control) and an experimental group consisting of the basal diet supplemented with 1 g of surfactin C and $1.0{\times}10^9CFU$ of Bacillus subtilis BC1212/kg feed. Piglets fed Bacillus subtilis BC1212 increased in average daily weight gain and feed efficiency. In comparison with the control group, the fecal Bacillus subtilis were significantly increased and the fecal coliform bacteria were markedly reduced in the experimental group. In addition, Bacillus subtilis BC1212 had excellent acid and bile tolerance. The treatment of surfactin($50{\mu}g\;ml^{-1}$) in lipopolysaccharide(LPS)-stimulated swine peripheral blood mononuclear cells(PBMCs) for 6 h showed a significant inhibitory effect on INF-$\gamma$, TNF-$\alpha$ and NO secretion(p<0.05) in comparison with LPS treatment alone but not on IL-10 secretion, with levels of secreted IL-10 similar to those secreted by PBMCs stimulated with LPS alone. Supplementation with surfactin in combination with Bacillus subtilis BC1212 in diets improved the ecosystem of gastrointestinal tract by increasing probiotic population and enhanced the systemic immune response in weaned piglets.

Potentials of Synbiotics for Pediatric Nutrition and Baby Food Applications: A Review (소아 영양 및 유아식 응용을 위한 신바이오틱스의 잠재력: 총설)

  • Jung, Hoo Kil;Kim, Sun Jin;Seok, Min Jeong;Cha, Hyun Ah;Yoon, Seul Ki;Lee, Nah Hyun;Kang, Kyung Jin
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.2
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    • pp.111-118
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    • 2015
  • Probiotic, prebiotic, and synbiotic substances as well as microorganisms were added to infant formula in an attempt to influence the intestinal microflora with an aim to stimulate the growth of lactic acid bacteria, especially bifidobacteria and lactobacilli. Over the last 10 years, new synbiotic infant formulas containing probiotics and prebiotics have been proposed in order to simulate the effect of breast-feeding on the intestinal microflora. Owing to their synergistic effect, the new synbiotics are expected to be more helpful than using probiotics and prebiotics individually. Maintenance of the viability of the probiotics during food processing and the passage through the gastrointestinal tract should be the most important consideration, since a sufficient number of bacteria ($10^8cfu/g$) should reach the intended location to have a positive effect on the host. Storage conditions and the processing technology used for the manufacture of products such as infant formula adversely affect the viability of the probiotics. When an appropriate and cost-effective microencapsulation methodology using the generally recognized as safe (GRAS) status and substances with high biological value are developed, the quality of infant formulas would improve. The effect of probiotics may be called a double-effect, where one is an immunomodulatory effect, induced by live probiotics that advantageously alter the gastrointestinal microflora, and the other comprises anti-inflammatory responses elicited by dead cells. At present, a new terminology is required to define the dead microorganisms or crude microbial fractions that positively affect health. The term "paraprobiotics" (or ghost probiotics) has been proposed to define dead microbial cells (not damaged or broken) or crude cell extracts (i.e., cell extracts with complex chemical composition) that are beneficial to humans and animals when a sufficient amount is orally or topically administered. The fecal microflora of bottle-fed infants is altered when the milk-based infant formula is supplemented with probiotics or prebiotics. Thus, by increasing the proportion of beneficial bacteria such as bifidobacteria and lactobacilli, prebiotics modify the fecal microbial composition and accordingly regulate the activity of the immune system. Therefore, considerable attention has been focused on the improvement of infant formula quality such that its beneficial effects are comparable to those of human milk, using prebiotics such as inulin and oligosaccharides and potential specific probiotics such as bifidobacteria, which selectively stimulate the proliferation of beneficial bacteria in the microflora and the indigenous intestinal metabolic activity of the microflora.

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Anti-Inflammatory Effect of Ethanol Extract from Onion (Allium cepa L.) Peel on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears (LPS로 유도된 RAW 264.7 세포와 마우스 귀 조직에 대한 양파(Allium cepa L.) 껍질 에탄올 추출물의 항염증 효과)

  • Ahn, Na-Kyung;Kang, Bo-Kyeong;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Bae, Nan-Young;Park, Ji-Hye;Park, Sun-Hee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1612-1620
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    • 2015
  • Inflammation is a complex process involving a variety of immune cells, which defend the body from harmful stimuli. However, pro-inflammatory cytokines and inflammatory mediators can also exacerbate diseases such as cancer. Onion peel contains several phenolic compounds, including quercetin at an amount 20 times greater in peel than edible flesh. Therefore, in this study, the anti-inflammatory effects of onion peel ethanol extract (OPEE) were investigated lipopolysaccharide-induced inflammatory response. In our results, NO production decreased in a dose-dependent manner. Secretion of IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ was suppressed by 44%, 53%, and 60% respectively, at $100{\mu}g/mL$. Moreover, OPEE also suppressed expression of COX-2, iNOS, $NF-{\kappa}B$, and MAPKs in a dose-dependent manner. Formation of mice ear edema was reduced at the highest dose tested compared to the control, and reduction of ear thickness was observed in the histological analysis as well. In the acute toxicity test, no morality was observed in mice administered 5,000 mg/kg body weight of OPEE over a 2-week observation period. These results suggest that OPEE may have significant effects on inflammatory factors and be a potential anti-inflammatory material.

Immunomodulatory and Anti-Inflammatory Activity of Mulberry (Morus alba) Leaves Fermented with Hericium erinaceum Mycelium by Solid-State Culture (Solid-State Culture를 이용하여 조제한 노루궁뎅이버섯 균사체-뽕잎발효물의 면역 및 항염증 활성)

  • Kim, Hoon;Jeong, Jae-Hyun;Shin, Ji-Young;Kim, Dong-Goo;Yu, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.9
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    • pp.1333-1339
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    • 2011
  • After mulberry (Morus alba) leaves were fermented with Hericium erinaceum mycelium by solid-state culture to enhance physiological activity, fermented mulberry leaves (MA-HE) was extracted by hot-water (MA-HEHW) and ethanol (MA-HE-E). MA-HE-HW showed enhanced mitogenic and intestinal immune system modulating activities (1.41 and 1.52 fold of saline control, respectively) compared to hot-water extracts of non-fermented mulberry leaves (MA-HW) and H. erinaceum mycelium (HE-HW) at $100\;{\mu}g$/mL. Meanwhile, when we tested the inhibitory effects of extracts on nitric oxide (NO), tumor necrosis factor (TNF)-${\alpha}$, and interleukin (IL)-$1{\beta}$ and IL-6 production, MA-HE-E significantly inhibited these pro-inflammatory mediators in LPS-stimulated RAW 264.7 cells (45.1, 41.3, 70.2, and 55.7% inhibition of LPS control at $1,000\;{\mu}g$/mL). In addition, MA-HE-HW and MA-HE-E did not show any cytotoxicity on RAW 264.7 cells at $1,000\;{\mu}g$/mL whereas HE-E and MA-E indicated cytotoxicity (80.1 and 30.7% cell viability of saline control). These results suggest that mulberry leaves fermented with H. erinaceum by solid-state culture might have enhanced immunomodulatory and anti-inflammatory effects compared to non-fermented mulberry leaves, resulting in ingredients biotransformed for fermentation with H. erinaceum mycelium.

Characterization and Distribution of Glycoconjugates in Human Pulmonary Tubercles by Lectin Histochemistry (폐결핵 결절에서 복합당질의 분포에 관한 Lectin 조직화학적 연구)

  • Yoon, Sik;Kim, Ji-Hong;Shin, Cheol-Shik;Jeong, Suk;Son, Mal-Hyun;Song, Sun-Dae;Kim, Jin-Jeong
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.3
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    • pp.248-261
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    • 1994
  • Background: Lectins are proteins or glycoproteins of non-immune origin that recognize a specific sequence of sugar residues. The availability of a large number of lectins has provided the capacity to identify selectively glycoconjugates possessing distinctive chemical structure in diverse sites of highly specialized biological activity. The purpose of the present study was to investigate the lectin binding patterns of various components in human pulmonary tubercles. Method: Biopsy specimens of tuberculous lung were obtained from male adult patients who underwent a surgical resection for severe pulmonary tuberculosis. The specimens were processed and stained with 13 kinds of biotinylated lectins according to some modification of Hsu and Raine's methods. Results: 1) In the caseous necrotic lesions, BS $I-B_4$ showed negative reaction and BS I were also negative except some irregularly-shaped cells located in the marginal zone. All other lectins, however, showed a positive reaction with various binding patterns. 2) The epithelioid cells were broadly divided into three groups according to the reaction patterns in the cytoplasms and cell membranes. 3) WGA, ECL, PHA-L, PHA-E and LCA showed strong staining in the lymphocytes. 4) SBA showed a different binding patterns between the endothelial layers located in the region beyond the fibrous layers and those located within the fibrous layers. 5) PNA showed a positive reaction in the outer 1/3 to 1/2 of the fibrous layer, but showed no staining in the inner 1/2 to 2/3 of the fibrous layers. Conclusion: The present lectin histochemical study provided a useful information to assess the characterization and distribution of various glycoconjugates in each constituent of human pulmonary tubercles. The results demonstrate structural differences in the glycoconjugate composition of various components of the tubercles and reveal changes in glycosylation in the components during soft tubercle formation. This study provides a new data useful for the studies on the pathogenesis and pathology of human pulmonary tubercles.

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Pharmacological Activity of Chaga Mushroom on Extraction Conditions and Immunostimulating Polysaccharide (추출조건에 따른 차가버섯 생리활성 및 면역활성 다당)

  • Baek, Gil-Hun;Jeong, Heon-Sang;Kim, Hoon;Yoon, Taek-Joon;Suh, Hyung-Joo;Yu, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.10
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    • pp.1378-1387
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    • 2012
  • To investigate the pharmacological activity of chaga mushroom (Inonotus obliquus) on extraction conditions, chaga was extracted using water (reflux at $50^{\circ}C$, decoction over $90^{\circ}C$, pressure at $121^{\circ}C$) or ethanol (reflux at 50, 70, or $90^{\circ}C$). When water extract was further fractionated into crude polysaccharide (IO-CP), yields of IO-CP (4.8~16.8%) were higher than those of ethanolic extracts (IO-E, 1.9~2.7%) at increased temperature. For antioxidant activity, crude polysaccharide (IO-CP-121) obtained by pressurized extraction showed the highest polyphenolic and flavonoid contents (35.10 mg TAE/g and 18.48 mg QE/g, respectively) as well as DPPH and ABTS free radical scavenging activities (26.08 and 27.99 mg AEAC/100 mg, respectively). Meanwhile, IO-CP-D (decoction) and IO-CP-50 (reflux) had more potent mitogenic effects (2.10- and 1.95-fold of saline control at 100 ${\mu}g/mL$) as well as intestinal immune system modulating activities (6.30- and 5.74-fold) compared to IO-CP-121, whereas ethanolic extracts showed no activity. Although no IO-CP showed cytotoxicity against RAW 264.7 cells at 0.1 mg/mL, IO-CP-121 significantly inhibited TNF-${\alpha}$ and NO production as pro-inflammatory factors in LPS-stimulated RAW 264.7 cells (29.2 and 63.5%, respectively). Ethanolic extracts also showed no cytotoxicity at 0.1 mg/mL, whereas inhibition of TNF-${\alpha}$ and NO production was significantly low compared to that of IO-CP-121. In addition, active IO-CP-D was further fractionated into an unadsorbed (IO-CP-I) and seven adsorbed fractions (IO-CP-II~VIII) by DEAE-Sepharose CL-6B column chromatography in order to isolate immunostimulating polysaccharide. IO-CP-II showed the most potent mitogenic effect and macrophage stimulating activity (4.51- and 1.64-fold, respectively). IO-CP-II mainly contained neutral sugars (61.86%) in addition to a small amount of uronic acid (2.96%), and component sugar analysis showed that IO-CP-II consisted mainly of Glc, Gal, and Man (molar ratio of 1.00:0.55:0.31). Therefore, extraction conditions affect the physiological activity of chaga, and immunostimulating polysaccharide fractionated from chaga by decoction is composed mainly of neutral sugars.

THE ROLE OF TYPE 2 DIABETES AS A PREDISPOSING RISK FACTOR ON THE PULPO-PERIAPICAL PATHOGENESIS: REVIEW ARTICLE (치수 치근단 병소의 전구 위험요인으로서의 제 2 형 당뇨의 역할에 관한 소고)

  • Kim, Jin-Hee;Bae, Kwang-Shik;Seo, Deog-Gyu;Hong, Sung-Tae;Lee, Yoon;Hong, Sam-Pyo;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.34 no.3
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    • pp.169-176
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    • 2009
  • Diabetes Mellitus (DM) is a syndrome accompanied with the abnormal secretion or function of insulin, a hormone that plays a vital role in controlling the blood glucose level (BGL). Type land 2 DM are most common form and the prevalence of the latter is recently increasing, The aim of this article was to assess whet her Type 2 DM could act as a predisposing risk factor on the pulpo-periapical pathogenesis. Previous literature on the pathologic changes of blood vessels in DM was thoroughly reviewed. Furthermore, a histopathologic analysis of artificially-induced periapical specimens obtained from Type 2 diabetic and DM-resistant rats was compared. Histopathologic results demonstrate that the size of periapical bone destruction w as larger and the degree of pulpal inflammation was more severe in diabetic rats, indicating that Type 2 D M itself can be a predisposing risk factor that makes the host more susceptible to pulpal infection. The possible reasons may be that in diabetic state the lumen of pulpal blood vessels are thickened by atheromatous deposits, and microcirculation is hindered, The function of polymorphonuclear leukocyte is also impair ed and the migration of immune cells is blocked, leading to increased chance of pulpal infection. Also, lack of collateral circulation of pulpal blood vessels makes the pulp more susceptible to infection. These decrease the regeneration capacity of pulpal cells or tissues, delaying the healing process, Therefore, when restorative treatment is needed in Type 2 DM patients, dentists should minimize irritation to the pulpal tissue un der control of BGL.

Effect of Jehotang Extract on the Growth of Intestinal Bacteria and Immunostimulation (제호탕의 장내 세균 및 면역 활성에 미치는 연구)

  • Ji, Myoung-Soon;Park, Min-Jung;Lee, Mi-Young;Kim, Jong-Goon;Ko, Byoung-Seob
    • Korean Journal of Food Science and Technology
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    • v.38 no.1
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    • pp.104-108
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    • 2006
  • Water extracts of Jehotang were evaluated for their growth-promoting effects on Bifidobacterium longum, Lactobacillus sp., L. acidophilus, and Clostridium perfringens. Addition of Jehotang water extract to modified EG media at 0.1 mg/mL increased growths of B. longum, Lactobacillus sp., and L. acidophilus, with 1.8-fold increase in growth of L. acidophilus compared to that of control. Studies on these strains by agar diffusion method showed Lactobacillus sp. and L. acidophilus were activated by addition of Jehotang extract at 10 mg/disc. Proliferation responses of mice splenocytes and Peyer's patch cells to ConA by LPS-stimulation at 500 mg/kg B.W./day Jehotang extract were investigated in vitro. Upon treatment of 1 mg/mL Jehotang water extract to mice, proliferations of splenocytes and Peyer's patch cells increased 1.4- and 1.6-fold compared to control, respectively. In mice administered Jehotang extract, production of intestinal secretory IgA (sIgA) increased 2.4-fold compared to control. These results indicate water extract of Jehotang stimulated intestinal immune system of mice. In mice treated with Jehotang extract, production of lymphocytes was 4% lower, whereas those of granulocytes and platelets were 4% and slightly higher than control, respectively.