• 대한한의학회지
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• 제31권5호
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• pp.112-123
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• 2010

Objectives: Three yin-tonifying formulae (Ssanghwa-tang, Yukmijihwang-tang and Jaeumganghwa-tang) were applied to investigate their immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in the murine macrophage cell line (RAW 264.7) and in ovalbumin (OVA)-immunized mice. Methods: This study was carried out in nitricoxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes in association with three herbal formulas. C57BL/6 mice were immunized intraperitoneally with OVA/aluminum ($100\;{\mu}g/200\;{\mu}g$/mouse) on days 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. Results: All three yin-tonifying formulas significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<0.001). Also, these herbal formulas all significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. Conclusion: These results suggested that the three yin-tonifying formulae could be used as stimulators of immune response.

• The Korean Journal of Physiology and Pharmacology
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• 제10권1호
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• pp.19-24
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• 2006

The intermediate conductance $Ca^{2+}-activated$ $K^+$ channels (SK4, IKCa1) are present in lymphocytes, and their membrane expression is upregulated by various immunological stimuli. In this study, the activity of SK4 was compared between Bal-17 and WEHI-231 cell lines which represent mature and immature stages of murine B lymphocytes, respectively. The whole-cell patch clamp with high-$Ca^{2+}$ ($0.8{\mu}M$) KCl pipette solution revealed a voltage-independent $K^+$ current that was blocked by clotrimazole (1 mM), an SK4 blocker. The expression of mRNAs for SK4 was confirmed in both Bal-17 and WEHI-231 cells. The density of clotrimazole-sensitive SK4 current was significantly larger in Bal-17 than WEHI-231 cells ($-11.4{\pm}3.1$ Vs. $-5.7{\pm}1.15$ pA/pF). Also, the chronic stimulation of B cell receptors (BCR) by BCR-ligation (anti-IgM Ab, $3{\mu}g$/ml, 8∼12 h) significantly upregulated the amplitude of clotrimazolesensitive current from $-11.4{\pm}3.1$ to $-53.1{\pm}8.6$ pA/pF in Bal-17 cells. In WEHI-231 cells, the effect of BCR-ligation was significantly small ($-5.7{\pm}1.15$ to $-9.0{\pm}1.00$ pA/pF). The differential expression and regulation by BCR-ligation might reflect functional changes in the maturation of B lymphocytes.

• 사상체질의학회지
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• 제22권4호
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• pp.77-84
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• 2010

1. Objectives Yanggyuksanhwa-tang for Soyangin was applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses on murine macrophage cell line (RAW 264.7) and ovalbumin/aluminium (OVA/Alum)-immunized mice. 2. Methods This study were carried out in nitric oxide (NO) synthesis on RAW 264.7 cells and cellular proliferation on mouse splenocytes. C57BL/6 mice were immunized intraperitonially with OVA/Alum (100 ${\mu}g$/200 ${\mu}g$) on day 1, 8, and 15. Yanggyuksanhwa-tang was administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. 3. Results Yanggyuksanhwa-tang significantly enhanced cellular proliferation by LPS and Con A on splenocytes from OVA/Alum-immunized mice (p<.001). Yanggyuksanhwa-tang also significantly enhanced plasma OVA-specific IgG (p<.001), IgG1 (p<.001), and total IgM (p<.01) levels compared with the OVA/Alum group. 4. Conclusions These results suggested that Yanggyuksanhwa-tang for Soyangin could be used as immunopotent.

• Archives of Pharmacal Research
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• 제20권2호
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• pp.128-137
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• 1997

The effects of cylindan, a polysaccharide isolated from the basidiocarps of Agrocybe cylindracea, on murine sarcoma 180 tumor and murine immune cells were examined after intraperitoneal administration. Cylindan exhibited a marked life extension effect in mice against ascite forms of sarcoma 180 and Lewis lung carcinoma at a dose of 50 mg/kg/day, although it did not show any direct cytotoxicity against sarcoma 180, X5563, and MM46 murine tumor cells. Cylindan increased numbers of bone marrow stem cells as well as peritoneal exudate cells in flow cytometry using monoclonal antibodies. The tumor bearing mice group apparently showed the increase of macrophages and cytotoxic T lymphocytes in mouse spleen cells during the early stage of tumor growth. But during the later stage, the control group decreased immune cells and cylindan restored the decreased immune cells in the tumor bearing mice to the normal level. In non-specific immune response, cylindan stimulated the bacterial phagocytosis and acid phosphatase production in macrophages. It also activated components of the alternative complement pathway and natural killer activity against YAC-1 lymphoma. In number of plasma cells as token of stimulation of the differentiation of B lymphocytes. In cellular immunity, cylindan restored the depressed response of delayed type hypersensitivity in the tumor bearing mice to 60% of the normal level and increased the interleukin-2 (IL-2) responsiveness in the IL-2 dependent CTLL-2 cells. These results suggest that cylindan did not show direct cytotoxic effects on tumor cells but restored the decreased immune response of the tumor bearing mice.

• Journal of Veterinary Science
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• 제25권1호
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• pp.4.1-4.14
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• 2024

Background: Lawsonia intracellularis is the causative agent of proliferative enteropathy and is associated with several outbreaks, causing substantial economic loss to the porcine industry. Objectives: In this study, we focused on demonstrating the protective effect in the mouse model through the immunological bases of two vaccine strains against porcine proliferative enteritis. Methods: We used live-attenuated Salmonella Typhimurium (ST) secreting two selected immunogenic LI antigens (Lawsonia autotransporter A epitopes and flagellin [FliC]-peptidoglycan-associated lipoprotein-FliC) as the vaccine carrier. The constructs were cloned into a Salmonella expression vector (pJHL65) and transformed into the ST strain (JOL912). The expression of immunogenic proteins within Salmonella was evaluated via immunoblotting. Results: Immunizing BALB/c mice orally and subcutaneously induced high levels of LI-specific systemic immunoglobulin G and mucosal secretory immunoglobulin A. In immunized mice, there was significant upregulation of interferon-γ and interleukin-4 cytokine mRNA and an increase in the subpopulations of cluster of differentiation (CD) 4⁺ and CD 8⁺ T lymphocytes upon splenocytes re-stimulation with LI antigens. We observed significant protection in C57BL/6 mice against challenge with 10^6.9 times the median tissue culture infectious dose of LI or 2 × 10⁹ colony-forming units of the virulent ST strain. Immunizing mice with either individual vaccine strains or co-mixture inhibited bacterial proliferation, with a marked reduction in the percentage of mice shedding Lawsonia in their feces. Conclusions: Salmonella-mediated LI gene delivery induces robust humoral and cellular immune reactions, leading to significant protection against LI and salmonellosis.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.103-108
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• 1996

Through the previous studies(I,II), it was observed that human follicular fluid(HFF) was more effective than human fetal cord serum(HFCS) on promoting meiotic resumption of oocytes and improving embryonic development of mouse in vitro. On the basis of these results, we have gradually exchanged HFCS with HFF as protein supplement in human ART. This study was performed to investigate the efficiency of HFF on improving the pregnancy rate in ART. Oocytes were retrieved transvaginally from patients treated with pituitary suppression with GnRH-agonist and ovarian stimulation with human menopausal gonadotro-pin(HMG) and pure follicle stimulating hormone(FSH). Aspirated oocytes were rinsed and cultured in TCM-199 containing HFF, and the concentrations of HFF were adjusted to 10, 20, and 30% according to the use for insemination, embryo growth and embryo transfer, respectively. As possible as, we tried to do embryo transfer into fallopian tube to mimic the coincidence of the cell stage with the place of sojourn in vivo, so we performed various ART programs(IVF & ET; in vitro fertilization, ZIFT; zygote intra fallopian-tube transfer, ZIFT & ET) according to the tubal conditions of patients. On the while, intra cytoplasmic sperm injection(ICSI) was used to assist IVF of the patients who had shown poor standard IVF results by immunological or severe male factor. Of the 255 cycles of ART programs using HFF as protein supplement, 118 cycles were turn out to be succeeded in pregnancy(46.2%, per cycle, p<0.05), while 21 pregnancies were achieved in the 69 cycles using HFCS(30.4%). The 255 cycles using HFF were subdivided into cycles with the type of ART programs, and each pregnancy rate of the ART programs were 44.7% (IVF & ET, 76/170 cycles), 53.4%(ZIFT, 31/58 cycles) and 40.7% (ZIFT & ET, 11/27 cycles), respectively. In the 61 ICSI cycles using HFF, 28 cycles succeed in pregnancy(45.9%), while 7 pregnancies were obtained in the 17 ICSI cycles using HFCS. Also the ongoing pregnancy rate in the group using HFF(78.8%, 93/118 cycles) was higher than that in the group using HFCS(61.9%). Therefore, we found that the use of HFF as protein supplement was more suitable and effective than the use of HFCS to improve the pregnancy rate in ART.

• 한국식품영양과학회지
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• 제33권3호
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• pp.469-474
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• 2004

본 연구에 사용한 아라비녹실란화합물은 미강에서 추출한 헤미셀룰로즈에 표고버섯균사체 배양물에서 분리한 효소를 작용시켜 생성된 식물성다당류이며, PSP(polysaccharide peptide)는 담자균류의 일종인 Coliolus versicocor Sov-1의 균사체에서 추출 정제하여 얻은 단백다당체 성분이다. 이들에 대한 면역세포 활성화 효과, 항암 효과 및 알레르기 억제효과 등에 관한 실험 결과는 다음과 같다. 비장세포의 B cell과 T cell에 arabinoxylane과 PSP의 항체생성과 세포활성능을 조사한 결과 PFC형 성은 10 ∼ 15%, RFC검출은 10 ∼ 30% 증가하였다 복강침출세포로부터 대식세포의 탐식능 효과는 대조군에 비하여 PSP는 약 18%, arabinoxylane은 22% 정도 증가하였다. S-180 고형암에 의한 항암실험에서 arabinoxylane은 66.2%(p<0.01), PSP는 49.0%(p<0.05)의 억제 효과를 확인할 수 있었다. 복수암에 의한 수명 연장효과는 PSP, arabinoxylane이 대조군에 비해 다소의 수명연장효과가 나타났으며, PSP가 아라비녹실란에 비해 약간의 높은 수명연장효과가 나타났다. 항알레르기 효과를 보기 위한 PCA 역가를 측정한 결과는 대조군이 80인데 비해 PSP군은 40, arabinoxylane군은 20으로 대조군에 비하여 다소 경감되었으며, arabinoxylane군이 다소 우수한 결과를 보였다. 혈중 histamine 함량도 PCA와 유사한 결과로 arabinoxylane군과 PSP군이 히스타민 함량을 다소 감소시키는 경향을 보였다.

• 한국식품영양과학회지
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• 제33권2호
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• pp.278-286
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• 2004

Arabinoxylane과 PSP는 농도 의존적으로 비장세포의 증식과 IFN-${\gamma}$ 생산을 유도하였으며 특히, PSP는 IL-2, IL-4, IL-10의 생산도 유도하였다. 그리고, 항체생산 세포수를 나타내는 PFC은 11∼14%, 활성화된 T세포수를 나타내는 RFC은 9∼30% 증가하였다. Arabinoxylane은 T세포의 증식을 유도하지 못하였지만, PSP는 높은 농도에서 T세포의 증식반응을 유도하였다 Arabinofylane과 PSP는 비장세포에서 분리한 B세포의 증식을 유도하였고, 대식세포의 탐식능은약 20∼22%정도 증가하였다. 그리고, 농도 의존적으로 대식세포의 일산화질소 생산을 유도하였으며, TNF-$\alpha$와 IL-6의 생산을 현저하게 유도하였다. 특히, PSP는IL-1$\beta$의 생산도 유도하였다. 또한, 대식세포 표면의 주조직적합 복합체II(MHC Class II)의 발현을 유도하였다. 이상의 결과로 arabinoxylane과 PSP는 면역반응에 관여하는 T세포, B세포 그리고 대식 세포의 다양한 활성을 유도하는 효과가 있는 것으로 생각되어 면역기능을 강화하는 소재와, 노약자나 회복기 환자 등을 위한 특수 식이용 소재로서 사용이 가능한 것으로 생각된다.

• 생명과학회지
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• 제22권6호
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• pp.828-836
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• 2012

일반적인 버섯 배지(미강 10%), 폐 한방슬러지를 10% 첨가한 배지, 그리고 미강 10%와 폐 한방슬러지 10%를 혼합한 배지에서 재배한 팽이버섯의 면역기능 강화효과를 알아보기 위해 물 추출물과 에탄올 추출물로 분리하여 실험하였다. 그 중 폐 한방슬러지 10%를 첨가한 배지에서 재배한 팽이버섯의 에탄올 추출물을 첨가하였을 때, 비장세포의 증식을 크게 유도하였으며 또한 IL-6, TNF-${\alpha}$, IFN-${\gamma}$ 분비를 유도하였다. 그리고 B세포의 증식반응과 면역글로불린 생산도 증가하였으며, 대식세포주의 일산화질소 분비와 IL-6, TNF-${\alpha}$, GM-CSF 생산도 증가하였다. 또한 복수암 유발 종양세포를 이용한 항종양 효과를 측정 한 결과 대조군의 평균 21.1일보다 실험군은 24.5일로 16.1%의 수명 연장효과가 나타났다. 따라서 폐 한방슬러지를 이용하면 수입에 의존하고 있는 버섯 재배 배지 원료를 절약할 수 있으며 면역세포조절 기능을 강화시키는 기능성 버섯을 얻을 수 있다고 생각한다.

• Clinical and Experimental Pediatrics
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• 제48권6호
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• pp.634-639
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• 2005

목 적 : 2세 미만의 소아에서는 임상적으로 확실한 증상이 있거나 알레르기 가족력이 있는 것을 제외하고는 실제적으로 아토피 유무를 판별하는 것은 성인에 비해 제한점이 많다. 본 연구에서는 영유아기에 알레르기 질환과 관련된 사이토카인이 유전자 배열에서 발현하는 지를 말초혈액의 Th2 림프구에서 interleukin-4 유전자의 DNA 탈메틸화를 통하여 알아보고자 하였다. 방 법 : 대상은 한양대학교 구리병원 소아알레르기클리닉을 내원한 3세 미만 소아에 대해 소아의 병력과 가족력을 조사하여 family allergy score와 개인력을 기준으로 알레르기질환에 대한 고위험군 7명, 저위험군 8명, 정상 대조군 7명이었다. 이들 모두에서 총 IgE치와 Der f II 특이 IgE를 측정하였고 이들의 말초혈액에서 T세포를 Der f II로 자극하여 배양한 후 DNA 추출을 하여 IL-4 유전자 두 번째 인트론의 CpG 섬에 대한 메틸화 분석을 실시하였다. 결 과 : 정상군에 비해 알레르기 환아군에서 IL4 유전자의 두번째 인트론에 위치한 CpG 섬의 두 부위 모두에서 메틸화 특이 PCR 신호강도가 약하게 나타나는 반면 정상아에서는 118 bp PCR 산물과 205 bp PCR 산물이 모두 강하게 나타나서 메틸화된 것으로 보이며, 저위험군 알레르기 환아에서 최소한 한 부위에서 탈메틸화가 일어났으며 고위험군 알레르기 환아에서는 두 부위 모두에서 탈메틸화가 일어났다. 결 론 : 정상 대조군의 T 세포에서는 IL-4 유전자의 메틸화가 일어나고 알레르기 환아에서 고위험군과 저위험군 모두 정도의 차이는 있으나 메틸화의 소실이 관찰되었다. 이를 이용하여 향후 영유아기에서 아토피 유무를 판별할 수 있는 가능성을 제시하였으나 더 많은 연구가 필요할 것으로 사료된다.