• IMMUNE NETWORK
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• 제3권3호
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• pp.188-200
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• 2003

Background: Immunization of dendritic cells (DCs) pulsed with tumor antigen can activate tumor-specific cytotoxic T lymphocytes (CTL) that are responsible for protection and regression. In this study, we examined whether the uptake of necrotic tumor cells could modulate DC phenotypes and whether the immunization of necrotic tumor cell-loaded DCs could elicit efficient tumor specific immune responses followed by a regression of established tumor burdens. Methods: We prepared necrotic tumor cell-pulsed DCs for the therapeutic vaccination and investigated their phenotypic characteristics, the immune responses induced by these DCs, and therapeutic vaccine efficacy against colon carcinoma in vivo. Several parameters including phagocytosis of tumor cells, surface antigen expression, chemokine receptor expression, IL-12 production, and NK as well as CTL activation were assessed to characterize the immune response. Results: DCs derived from mouse bone marrow efficiently phagocytosed necrotic tumor cells and after the uptake, they produced remarkably increased levels of IL-12. A decreased CCR1 and increased CCR7 expression on DCs was also observed after the tumor uptake, suggesting that antigen uptake could induce DC maturation. Furthermore, co-culturing of DCs with NK cells in vitro enhanced IL-12 production in DCs and IFN-${\gamma}$ production in NK cells, which was significantly dependent on IL-12 production and cell-to-cell contact. Immunization of necrotic tumor cell-loaded DCs induced cytotoxic T lymphocytes as well as NK activation, and protected mice against subsequent tumor challenge. In addition, intratumoral or contra-lateral immunization of these DCs not only inhibited the growth of established tumors, but also eradicated tumors in more than 60% of tumor-bearing mice. Conclusion: Our data indicate that production of IL-12, chemokine receptor expression and NK as well as CTL activation may serve as major parameters in assessing the effect of tumor cell-pulsed DC vaccine. Therefore, DCs loaded with necrotic tumor cells offer a rational strategy to treat tumors and eventually lead to prolonged survival.

• 한국식품영양과학회지
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• 제43권9호
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• pp.1317-1324
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• 2014

본 연구에서는 울금 20% 주정 추출물이 LP-BM5 MuLV에 감염된 MAIDS 모델에서 면역조절 효과를 확인하기 위해 cytokines 생산, 혈장 면역글로불린 농도, T 세포 및 B 세포의 증식능, NK 세포의 활성능을 측정하였다. 그 결과 LPBM5 MuLV 감염으로 인하여 감소된 T 세포 및 B 세포의 증식능, NK 세포의 활성능을 울금 20% 주정 추출물 식이 투여가 증가시켰으며, Th1 type cytokines(IL-2, IFN-${\gamma}$)의 생성량을 증가시키고 Th2 type cytokines(IL-4, IL-10)은 억제시킴으로써 Th1/Th2 type cytokine 발현을 조절하여 면역 항상성을 유지하는 효과를 보였다. 따라서 울금은 면역조절 효과를 가진 천연 기능성 소재로서의 가능성을 기대할 수 있다.

• 한국식품영양과학회지
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• 제36권3호
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• pp.269-275
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• 2007

불가사리 추출물을 이용하여 생쥐 비장에 있는 면역세포 활성화 효과에 대해 실험한 결과, 다음과 같은 결과를 얻었다. 첫째, 별불가사리 추출물 중에서 B세포와 대식세포를 활성화시키는 성분은 아세톤처리로 추출할 수 있었다. 둘째, 이 성분은 농도 의존적으로 생쥐 비장세포의 증식반응을 유도하였으며, IL-6와 $IFN-{\gamma}$의 생산을 유도하였다. 셋째, 이 성분은 B세포의 증식을 유도하였으며, 이 때 면역글로불린 IgM과 IgG의 생산도 유도하였다. 넷째, 이 성분은 대식세포주의 일산화질소 생산을 유도하였으며, 또 $TNF-{\alpha}$, GM- CSF와 IL-6의 분비를 유도하였다. 이상의 실험 결과, 본 실험에서 사용한 별불가사리 추출물에는 B세포와 대식세포 같은 면역세포의 증식과 각종 사이토카인을 생산을 유도하여, 면역반응을 조절하는 성분이 포함되어 있는 것으로 생각된다.

• 한국식품영양과학회지
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• 제44권3호
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• pp.307-313
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• 2015

본 연구에서는 국내에서 재배되는 울금의 면역조절 효과에 대해 평가하고자 20% 주정 추출물을 이용하여 자연살해세포와 T 세포에 미치는 영향을 관찰하였다. 마우스의 비장세포에서 분리한 자연살해세포를 종양세포 YAC-1 세포와 함께 배양시켜 울금 20% 주정 추출물의 처리에 따른 변화를 관찰하였다. 그 결과 울금 20% 주정 추출물의 처리는 자연 살해세포의 CD69 발현과 IFN-${\gamma}$의 발현을 증가시켰고 결과적으로 활성이 증가되어 YAC-1 세포의 제거를 증가시켰음을 확인하였다. 또한 마우스의 비장세포에서 울금 20% 주정 추출물의 처리에 따른 T 세포의 변화를 관찰한 결과에서는 CD4+ T 세포보다는 CD8+ T 세포를 증가시켰음을 확인하여 감염된 세포나 종양세포 제거를 효과적으로 할 수 있을 것이라고 예상할 수 있었다. 결론적으로 울금 20% 주정 추출물은 내재면역과 적응면역에 영향을 미쳐 면역조절에 긍정적인 변화를 보였음을 확인하였다.

• 대한한의학방제학회지
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• 제21권1호
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• pp.51-70
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• 2013

Objectives : The object of this study was to observe anticancer and related immunomodulatory effects of Insamyangyoung-tang extracts (ISYYTe) on non-small cell lung carcinoma (squamous epithelial carcinoma), NCI-H520, xenograft Balb/c nu-nu nude mice. Methods : Three different dosages of ISYYTe, 50, 100 and 200 mg/kg were orally administered once a day for 42 days from 11 days after tumor cell inoculation. Six groups, which are intact control, tumor bearing control, 5-fluorouracil (FU) 30 mg/kg, ISYYTe 50 mg/kg, ISYYTe 100 mg/kg, ISYYTe 200 mg/kg, each of 8 mice per group were used in the present study. Changes on the body weight, tumor volume and weight, lymphatic organ (spleen and popliteal lymph node), serum interferon (IFN)-${\gamma}$ levels, splenocytes NK cell activity and peritoneal macrophage activities, splenic tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$ and IL-10 contents were observed with tumor mass and lymphatic organ histopathology to detect anticancer and immunomodulatory effects. Results : As results of ISYYTe 50, 100 and 200 mg/kg treatment, decreases in the tumor volumes and weights were detected. At histopathological observations, decreases of tumor cell volumes in tumor masses were dose-dependently decreased mediated by increases of apoptosis among tumor cells by treatment of all three different dosages of ISYYTe. As results of tumor cell inoculation, marked decreases of spleen and popliteal lymph node weights, serum IFN-${\gamma}$, splenic TNF-${\alpha}$, IL-$1{\beta}$ and IL-10 contents and splenocytes were observed with histopathological atrophic changes of spleen and popliteal lymph nodes. Conclusions : Over 50 mg/kg of ISYYTe showed favorable anticancer effects on the NCI-H520 cell xenograft with immunomodulatory effects. Although relatively lower anticancer effects were observed in ISYYTe 200 mg/kg treated mice as compared with 5-FU 30 mg/kg treated mice, there are no meaningful favorable immunomodulatory effects were observed after 5-FU treatment in the present study.

• 대한한방소아과학회지
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• 제30권4호
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• pp.29-59
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• 2016

Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

• 한국자원식물학회지
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• 제27권5호
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• pp.567-575
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• 2014

Resveratrol은 천연 stilbene으로 안전성 있는 항염증 활성을 가진 화합물로 알려져 있다. 최근의 연구들에서 resveratrol이 천식, 만성 대장염, 류마티스성 관절염과 같이 염증에 의해 발생하는 다양한 질병을 억제한다고 보고되었다. 이러한 연구들은 resveratrol이 $CD4^+$ helper T cells (Th cells)에 의한 면역반응을 조절할 것이라고 제시하였다. 그러나 resveratrol이 직접적으로 Th cells의 활성화와 분화를 조절하는지 완전히 밝혀지지 않았다. C57BL/6에서 Th cells을 분리하여 다양한 농도의 resveratrol을 세포에 처리하였다. 본 연구에서는 resveratrol이 직접적으로 Th cells의 활성화와 증식을 억제하는 것을 확인하였다. Th cells에 resveratrol을 처리하였을 때 IFN-${\gamma}$, IL-4, IL-17 사이토카인 생성이 농도에 따라 유의하게 감소하였고 또한 Th cells이 이러한 사이토카인들을 분비하는 Th1과 Th2과 Th17으로 분화되는 것이 억제되었다. 그리고 고농도의 resveratrol이 Th cells의 세포사멸을 유도하는 것으로 확인되었다. 본 연구에서는 resveratrol이 Th cells의 활성화와 분화를 직접적으로 억제하는 것을 확인하였으며, 이는 resveratrol이 $CD4^+$ Thcells에 의해 발생되는 자가면역질환의 효과적인 치료법이 될 수 있을 것이라고 제시한다.

• 한방안이비인후피부과학회지
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• 제26권1호
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• pp.19-34
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• 2013

Objective : This study was performed to evaluate the effect of immune reaction inductive substances such as PMA, LPS, DPE, DNCB and WP, the whitman pigra extracting substance at simultaneously on the translocation of $NF{\kappa}B$ towards to the nucleus and the mRNA expression patterns of various cytokine genes in THP-1 cells, monocytes of human. Methods : To analyze the cytokine genes expressions, the RT-PCR method was used, and measuring TNF-${\alpha}$ that had been secreted during cell culture by the ELISA method. The morphological changes were observed during THP-1 cell by a scanning electron microscope and the quantitative distribution of $NF{\kappa}B$ in the cell that was analyzed through immunocytochemistry and a confocal microscopy. Results : WP showed different influences onto the mRNA expression patterns of cytokine genes with PMA, LPS. DPE and DNCB according to the types of immune inductive substances in the THP-1 cells. Upon treating PMA and DPE on the THP-1 cells at the same time or either additionally treating WP thereon, the movement of $NF{\kappa}B$ increase towards the nucleus from cell cytoplasm was able to be observed. The expressions of IL-$1{\alpha}$ and IFN-${\gamma}$ induced by PMA and PMA+DNCB were suppressed by WP while the expression of TGF-${\beta}$ was promoted. Regarding the secretion pattern of TNF-${\alpha}$ according to the treatment of PMA, its secretion amount was incredibly increased by concurrent treatment of WP, however, in case of co-treatment of WP with PMA and DNCB, it was found that the secretion amount of TNF-${\alpha}$ decreased. Conclusions : In this study, the WP extracting substance was confirmed that it had an influence on expression patterns of cytokine genes according to the actions of a variety kinds of immune reaction inductive substances treated on the THP-1 cells. Especially, WP co-treatment with PMA and DNCB was suppressed the expression of inflammatory cytokines, such as IL-$1{\alpha}$, IFN-${\gamma}$ and TNF-${\alpha}$.

• Parasites, Hosts and Diseases
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• 제49권2호
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• pp.115-123
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• 2011

Toxoplasma gondii Korean isolate (KI-1) tachyzoites were inoculated intraduodenally to BALB/c mice using a silicon tube, and the course of infection and immune responses of mice were studied. Whereas control mice, that were infected intraperitoneally, died within day 7 post-infection (PI), the intraduodenally infected mice survived until day 9 PI (infection with $1{\times}10^5$ tachyzoites) or day 11 PI (with $1{\times}10^6$ tachyzoites). Based on histopathologic (Giemsa stain) and PCR (B1 gene) studies, it was suggested that tachyzoites, after entering the small intestine, invaded into endothelial cells, divided there, and propagated to other organs. PCR appeared to be more sensitive than histopathology to detect infected organs and tissues. The organisms spread over multiple organs by day 6 PI. However, proliferative responses of splenocytes and mesenteric lymph node (MLN) cells in response to con A or Toxoplasma lysate antigen decreased significantly, suggesting immunosuppression. Splenic $CD4^+$ and $CD8^+$ T-Iymphocytes showed decreases in number until day 9 PI, whereas IFN-${\gamma}$ and IL-10 decreased slightly at day 6 PI and returned to normal levels by day 9 PI. No TNF-${\alpha}$ was detected throughout the experimental period. The results showed that intraduodenal infection with KI-1 tachyzoites was successful but did not elicit significant mucosal immunity in mice and allowed dissemination of T. gondii organisms to systemic organs. The immunosuppression of mice included reduced lymphoproliferative responses to splenocytes and MLN cells to mitogen and low production of cytokines, such as IFN-${\gamma}$, TNF-${\alpha}$, and IL-10, in response to T. gondii infection.

• 한국수의병리학회지
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• 제3권1호
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• pp.15-25
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• 1999

To elucidate the mechanism of age-related development in FGS/NgaKIST mice with spontaneous glomerulosclerotic lesion, we examined expression and localization of various cytokine mRNA in the kidney in the progression of diseases. This mouse model is the first to develop spontanously occuring glomerosclerotic lesion in the kidney. In this study, we detected the up-regulation of local cytokine genes such as IL-1$\beta$, IL-2, IL-6, IL-10, TNF-$\alpha$, TGF-$\beta$, and IFN- $\gamma$ in the kidneys. In RT-PCR and Southern blot analysis, we detected gradual expressions of cytokine mRNA of IL-1$\beta$, IL-2, IL-6, IFN- $\gamma$, and TNF $\alpha$ mRNA during the course of disease. Other cytokines including IL -10 and TGF -$\beta$ were found to be appeared the slightly expressed level at 3 to 12 weeks before onset of inflammatory lesion but they are highly expressed at the end-stage of the disease accompaning high proteinurea and wasting. In situ RT-PCR, each cytokine mRNA were specifically localized in a variety of cells including mesangial, endothelial, parietal epithelial, tubular epithelial, arterial muscle cell, and infiltrated inflammatory cells. In addition, TNF - $\alpha$was detected moderately in the visceral and parietal epithelial cell, but weakly in endothelial and mesangial cells, whereas IL-1 $\beta$ and IL -6 were strong in mesangial regions. IL-6 and TNF- $\alpha$ was highly localized in the damaged proximal and collecting tubules. Especially, TGF -$\beta$ mRNA was highly found in mesangial cells within glomerulus and interstitium during the end-stage of this disease.. These results indicate that pro inflammatory cytokines such as IL-1 $\beta$, IL-2, IL-6, and TNF- $\alpha$ were gradually expressed from the early stage of this disease to the end-stage, and that IL-10 and TGF-$\beta$ may be important in the accumulation of extracellular matrix(ECM) within glomerulus and periglomerular fibrosis in the progression of this disease as well as tissue destruction in end-stage of this disease.