Ursolic acid isolated from Uncaria rhynchophylla activates human dendritic cells via TLR2 and/or TLR4 and induces the production of IFN-γ by CD4+ naive T cells

Jung, T.Y.,Pham, T.N.N.,Umeyama, A.,Shoji, N.,Hashimoto, T.,Lee, J.J.,Takei, M. North-Holland ; Elsevier Science Ltd 2010 european journal of pharmacology Vol.643 No.2

Ursolic acid is triterpene isolated from Uncaria rhynchophylla and is a pharmacologically active substance. The induction of dendritic cell maturation is critical for the induction of Ag-specific T-lymphocyte response and may be essential for the development of human vaccine relying on T cell immunity. In this study, we investigated that the effect of Ursolic acid on the phenotypic and functional maturation of human monocyte-derived dendritic cells in vitro. Dendritic cells harvested on day 8 were examined using functional assay. The expression levels of CD1a, CD80, CD83, CD86, HLA-DR and CCR7 on Ursolic acid-primed dendritic cells was slightly enhanced. Ursolic acid dose-dependently enhanced the T cell stimulatory capacity in an allogeneic mixed lymphocyte reaction, as measured by T cell proliferation. The production of IL-12p70 induced by Ursolic acid-primed dendritic cells was inhibited by the anti-Toll-like receptor-2 (TLR2) mAb and anti-TLR4 mAb. Moreover, Ursolic acid-primed dendritic cells expressed levels of mRNA coding for both TLR2 and TLR4. The majority of cells produced considerable interferon-gamma (IFN-γ), but also small amounts of interleukin (IL-4)-4. Ursolic acid-primed dendritic cells have an intermediate migratory capacity towards CCL19 and CCL21. These results suggest that Ursolic acid modulates human dendritic cells function in a fashion that favors Th1 polarization via the activation of IL-12p70 dependent on TLR2 and/or TLR4, and may be used on dendritic cells-based vaccines for cancer immunotherapy.

마우스 백혈병세포 RNA로 감작된 수상세포에 의한 세포독성 T 림프구의 유도

정철원,권정혜,설재구,박우현,현정미,김은실,김승택,이상재,김병국,이영열 大韓血液學會 2001 大韓血液學會誌 Vol.36 No.3

유선조직내에 출현하는 dendritic cell의 형태학적 연구 II. 전자현미경적 관찰

류시윤,이차수,Ryu, Si-yun,Lee, Cha-soo 대한수의학회 1988 大韓獸醫學會誌 Vol.28 No.2

In order to investigate the morphological characteristics of dendritic cells in the mammary gland of the mouse (C57 BL/6), rat(W), rabbit and cat, the fine structures of the dendritic cells have been observed by the electron microscope. The results obtained were summarized as follows: The dendritic cells with the well-developed processes had an irregular shape, and lacked the desmosome. The pinocytotic vesicles and tubular invaginations of the cell membrane were frequently observed, and the mitochondria with the well-developed cristae were located in the restricted region in the cytoplasm of the dendritic cells. The nuclei of dendritic cells were indented, In the mice and rats, the dendritic cells had a few Langerhans cell granules(Birbeck granules). From the above results, it is confirmed that the ATPase-positive dendritic cells in the mammary gland are the Langerhans cells.

The Modulation of Cell Wall-Associated Triton X-100 Soluble Protein (TSP) Antigens of Mycobacterium tuberculosis on Murine Dendritic Cells

Yung-Choon Yoo,Young-Joo Bae,Tae-Hyun Paik,Junglim Lee 한국실험동물학회 2008 Laboratory Animal Research Vol.24 No.4

Mycobacterium tuberculosis or its secreted protein antigens are known as a potent inducer of maturation process of dendritic cells, during which the expression of cell surface molecules and the capability of antigen presentation are increased, resulting in potent interaction with T cells to initiate the acquired immune response against tuberculosis. In this study, the cell wall-associated Triton X-100 soluble protein (TSP) antigens, TSP- BCG, TSP-H37Ra, TSP- H37Rv and TSP-K, were tested on murine bone marrowderived dendritic cells to determine the capability of their immunologic modulation. Only TSP-H37Rv stimulated murine dendritic cells showed moderate production of IL-6 and MIP-1α compared to those of LPS. TSP-H37Rv antigen did not induce the production of IL-12, TNF-α or IL-10. Other TSP antigenstimulated murine dendritic cells showed minimal or no induction of cytokines (IL-6, IL-12, TNF-α or IL-10) or chemokines (MIP-1α or RANTES). The murine dendritic cells showed the production of NO as well as the expression of iNOS mRNA after LPS stimulation, however, none of TSP antigen-stimulated murine dendritic cells induced either NO or iNOS mRNA expression. The allostimulation of T cells by murine dendritic cells, which were stimulated with each TSP antigens, showed insignificant increase in T cell proliferation activity in mixed lymphocyte reaction. These results suggested that cell wall-associated TSP antigen isolated from Mycobacterium tuberculosis H37Rv strain acts as a more potent stimulator on murine dendritic cells than any other TSP antigens, even the stimulation from TSP-H37Rv was not enough to activate the maturation of process of murine dendritic cells.

GM-CSF Grown Bone Marrow Derived Cells Are Composed of Phenotypically Different Dendritic Cells and Macrophages

Na, Yi Rang,Jung, Daun,Gu, Gyo Jeong,Seok, Seung Hyeok Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.10

Granulocyte-macrophage colony stimulating factor (GM-CSF) has a role in inducing emergency hematopoiesis upon exposure to inflammatory stimuli. Although GM-CSF generated murine bone marrow derived cells have been widely used as macrophages or dendritic cells in research, the exact characteristics of each cell population have not yet been defined. Here we discriminated GM-CSF grown bone marrow derived macrophages (GM-BMMs) from dendritic cells (GM-BMDCs) in several criteria. After C57BL/6J mice bone marrow cell culture for 7 days with GM-CSF supplementation, two main populations were observed in the attached cells based on MHCII and F4/80 marker expressions. GM-BMMs had $MHCII^{low}F4/80^{high}$ as well as $CD11c^+CD11b^{high}CD80^-CD64^+MerTK^+$ phenotypes. In contrast, GM-BMDCs had $MHCII^{high}F4/80^{low}$ and $CD11c^{high}CD8{\alpha}^-CD11b^+CD80^+CD64^-MerTK^{low}$ phenotypes. Interestingly, the GM-BMM population increased but GM-BMDCs decreased in a GM-CSF dose-dependent manner. Functionally, GM-BMMs showed extremely high phagocytic abilities and produced higher IL-10 upon LPS stimulation. GM-BMDCs, however, could not phagocytose as well, but were efficient at producing $TNF{\alpha}$, $IL-1{\beta}$, IL-12p70 and IL-6 as well as inducing T cell proliferation. Finally, whole transcriptome analysis revealed that GM-BMMs and GM-BMDCs are overlap with in vivo resident macrophages and dendritic cells, respectively. Taken together, our study shows the heterogeneicity of GM-CSF derived cell populations, and specifically characterizes GM-CSF derived macrophages compared to dendritic cells.

제대혈 CD34 양성세포를 이용한 수상돌기 세포의 배양

황보권,홍대식,김찬규,조성태,이규택,박성규,원종호,백승호,서원석,김숙자,박희숙 대한조혈모세포이식학회 1998 대한조혈모세포이식학회지 Vol.3 No.2

Background: Dendritic cells have a distinctive morphology and immunostimulatory potential. These cells are thought to be the major APC involved in triggering primary T-cell responses and express little linage specific surface markers. We tried to establish the method for the culture of dendritic cells with CD34^(+) cells from cord blood. Methods: Cord blood was collected during normal full-term delivery CD34^(+) cells were purified with Magnetic Separation Column, and cultured over 14 days in the presence of GM-CSF and TNF-α. Flow-cytometric analysis was conducted before and after culture. CFU-GM and CFU-DL culture was done with cells, harvested from 14 days of culture. Results: 1) After 14 days of culture with CD34^(+) cells (5×10^(5)), total cell number was expanded upto 3.2±3.35×10^(7), about 63 fold, and about 20% of them were adherent cells with dendritic morphology of numerals long and thin cytoplasmic projections. 2) Flow-cytometric analysis of surface marker expression were examined before and after culture for 14 days. The expression of CD34 was 69.1±13.01 (%), initially, and was decreased to 0.5±0.41 (%) at 14th day. The expression of HLA-DR, CD14 and CD1a were increased to 82.4±11.47 (%), 39.8±30.00 (%) and 15.7±7.73 (%) from 72.9±14.79 (%), 3.4±0.25 (%) and 1.5 ±0.64 (%). 3) The ratio of CFU-GM (94.5±86.97) and CFU-DL (8±8.49) colonies were about 10 to 1. Conclusion: CD34^(+) enriched cord blood can be expanded upto 63 folds with GM-CSF and TNF-α. This study established the method for the culture and identification of dendritic cells morphologically and immunophenotypically. Clinical application of dendritic cells as an adoptive immunotherapy and therapeutic vaccination in infectious or malignant disease needs further investigations about the right time of exposure to antigens, the dosage of cells, and the root of administration of the dendritic cells.

GM-CSF Grown Bone Marrow Derived Cells Are Composed of Phenotypically Different Dendritic Cells and Macrophages

Seung-Hyeok Seok,Yi Rang Na,Daun Jung,Gyo Jeong Gu 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.10

Granulocyte-macrophage colony stimulating factor (GM-CSF) has a role in inducing emergency hematopoiesis upon exposure to inflammatory stimuli. Although GM-CSF generated murine bone marrow derived cells have been widely used as macrophages or dendritic cells in research, the exact characteristics of each cell population have not yet been defined. Here we discriminated GM-CSF grown bone marrow derived macrophages (GM-BMMs) from dendritic cells (GM-BMDCs) in several criteria. After C57BL/6J mice bone marrow cell culture for 7 days with GM-CSF supplementation, two main populations were observed in the attached cells based on MHCII and F4/80 marker expressions. GM-BMMs had MHCIIlowF4/80high as well as CD11c+CD11bhighCD80-CD64+MerTK+ phenotypes. In contrast, GM-BMDCs had MHCIIhighF4/80low and CD11chighCD8α-CD11b+CD80+CD64-MerTKlow phenotypes. Interestingly, the GM-BMM population increased but GM-BMDCs decreased in a GM-CSF dose-dependent manner. Functionally, GM-BMMs showed extremely high phagocytic abilities and produced higher IL-10 upon LPS stimulation. GM-BMDCs, however, could not phagocytose as well, but were efficient at producing TNF, IL-1, IL-12p70 and IL-6 as well as inducing T cell proliferation. Finally, whole transcriptome analysis revealed that GM-BMMs and GM-BMDCs are overlap with in vivo resident macrophages and dendritic cells, respectively. Taken together, our study shows the heterogeneicity of GM-CSF derived cell populations, and specifically characterizes GM-CSF derived macrophages compared to dendritic cells.

갑상선암 환자의 말초혈액 백혈구로부터 수지상세포의 유도분화

이대희 고신대학교(의대) 고신대학교 의과대학 학술지 2006 고신대학교 의과대학 학술지 Vol.21 No.1

Background : Recent studies suggest that immunization with autologous dendritic cells(DCs) result in protective immunity and rejection of established tumors in various human malignancies, It has been reported that a dense infiltration of dendritic cells correlates with a favorable prognosis in several types of cancer. The purpose of this study is to determine whether DCs are generated from peripheral blood monocytes by using cytokines such as Fit-3 ligand, granulocyte macrophage-colony stimulating factor (GM-CSF), IL-4, and TNF- a,and whether cytotoxic T cells activated against the medullary thyroid carcinoma(MTC) tissues by the DCs. Methods : Peripheral blood was obtained from 2 patients with MTC. DCs were established from mononuclear leukocytes by culturing in the presence of Flt-3 ligand,GM-CSF, I卜4, and TNF- a for 14 days. At day 14,the differentiated DCs was analyzed morphologically. The immunophenotypic features of DCs such as CDla,CD83, and CD86 were analyzed by Immunofluorelescence microscopy. At day 15, DCs were incubated with thyroid cancer tissues and normal thyroid tissues for 7 additional days,respectively. Results : DCs were generated from the peripheral blood mononuclear cells. The generated cells showed the typical morphology of DCs. Activated cytotoxic T lymphocytes (CTLs) were observed. DCs attached to the thyroid cancer tissues and the CTLs were attached to the MTC tissues on scanning electron microscope. Conclusion : We could differentiate DCs from the peripheral blood mononuclear cells,And the DCs activate the CTLs which able to attack the MTC tissues. These results suggest that DCs can be used as adjuvants for immunotherapy of MTC. And this study represent the basis for the develop of new therapeutic strategies not only in MTC but also in the other malignancies. Background : Recent studies suggest that immunization with autologous dendritic cells(DCs) result in protective immunity and rejection of established tumors in various human malignancies, It has been reported that a dense infiltration of dendritic cells correlates with a favorable prognosis in several types of cancer. The purpose of this study is to determine whether DCs are generated from peripheral blood monocytes by using cytokines such as Fit-3 ligand, granulocyte macrophage-colony stimulating factor (GM-CSF), IL-4, and TNF- a,and whether cytotoxic T cells activated against the medullary thyroid carcinoma(MTC) tissues by the DCs. Methods : Peripheral blood was obtained from 2 patients with MTC. DCs were established from mononuclear leukocytes by culturing in the presence of Flt-3 ligand,GM-CSF, I卜4, and TNF- a for 14 days. At day 14,the differentiated DCs was analyzed morphologically. The immunophenotypic features of DCs such as CDla,CD83, and CD86 were analyzed by Immunofluorelescence microscopy. At day 15, DCs were incubated with thyroid cancer tissues and normal thyroid tissues for 7 additional days,respectively. Results : DCs were generated from the peripheral blood mononuclear cells. The generated cells showed the typical morphology of DCs. Activated cytotoxic T lymphocytes (CTLs) were observed. DCs attached to the thyroid cancer tissues and the CTLs were attached to the MTC tissues on scanning electron microscope. Conclusion : We could differentiate DCs from the peripheral blood mononuclear cells,And the DCs activate the CTLs which able to attack the MTC tissues. These results suggest that DCs can be used as adjuvants for immunotherapy of MTC. And this study represent the basis for the develop of new therapeutic strategies not only in MTC but also in the other malignancies.

The Role of Plasmacytoid and Myeloid Dendritic Cells in Induction of Asthma in a Mouse Model and the Effect of a TLR9 Agonist on Dendritic Cells

모지훈,정영준,Tomoko Hayashi,이종대,Eyal Raz 대한천식알레르기학회 2011 Allergy, Asthma & Immunology Research Vol.3 No.3

Purpose: To determine the role of plasmacytoid dendritic cells (pDC) and myeloid dendritic cells (mDC) in priming effector T cells to induce allergy, and to evaluate the effect of immunostimulatory sequences (ISS, TLR9 agonist) on dendritic cells. Methods: Cultured mDC and pDC with/without ISS were injected intratracheally into sensitized Balb/C mice. Mice were sacrificed, and then pulmonary function tests, bronchoalveolar lavage (BAL), cell counts, and cytokine levels were evaluated. Migration of dendritic cells was also evaluated after ISS administration. Results: In mice injected with mDC, airway hyperresponsiveness, eosinophil counts, and Th2 cytokine levels in BAL increased with increasing numbers of mDC injected. However,in mice injected with pDC, none of these changed, suggesting poor priming of T cells by pDC. In addition, mDC pulsed with ISS inhibited asthmatic reactions, and ISS administration inhibited migration of DC to the lung. Conclusions: We suggest that pDC played a limited role in priming T cells in this asthma model and that mDC played a major role in inducing asthma. In addition, ISS inhibited migration of DC to the lung.

Regulation of Th2 Cell Immunity by Dendritic Cells

Na, Hyeongjin,Cho, Minkyoung,Chung, Yeonseok 한국조명·전기설비학회 2016 한국조명·전기설비학회 학술대회논문집 Vol. No.

<P>Th2 cell immunity is required for host defense against helminths, but it is detrimental in allergic diseases in humans. Unlike Th1 cell and Th17 cell subsets, the mechanism by which dendritic cells modulate Th2 cell responses has been obscure, in part because of the inability of dendritic cells to provide IL-4, which is indispensable for Th2 cell lineage commitment. In this regard, immune cells other than dendritic cells, such as basophils and innate lymphoid cells, have been suggested as Th2 cell inducers. More recently, multiple independent researchers have shown that specialized subsets of dendritic cells mediate Th2 cell responses. This review will discuss the current understanding related to the regulation of Th2 cell responses by dendritic cells and other immune cells.</P>