• Title/Summary/Keyword: mononuclear cell

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In vitro effects of monophosphoryl lipid A and Poly I:C combination on equine cells

  • Dong-Ha Lee;Eun-bee Lee;Jong-pil Seo ;Eun-Ju Ko
    • Journal of Veterinary Science
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    • v.24 no.3
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    • pp.37.1-37.14
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    • 2023
  • Background: Toll-like receptor (TLR) agonists have been used as adjuvants to modulate immune responses in both animals and humans. Objectives: The objective of this study was to evaluate the combined effects of the TLR 4 agonist monophosphoryl lipid A (MPL) and the TLR 3 agonist polyinosinic:polycytidylic acid (Poly I:C) on equine peripheral blood mononuclear cells (PBMCs), monocyte-derived dendritic cells (MoDCs), and bone marrow-derived mesenchymal stromal cells (BM-MSCs). Methods: The PBMCs, MoDCs, and BM-MSCs collected from three mixed breed horses were treated with MPL, Poly I:C, and their combination. The mRNA expression of interferon gamma (IFN-γ), interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-12p40, tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), and monocyte chemoattractant protein-1 (MCP-1) was determined using real-time polymerase chain reaction. Results: The combination of MPL and Poly I:C significantly upregulated immunomodulatory responses in equine cells/ without cytotoxicity. The combination induced greater mRNA expression of pro-inflammatory cytokines IFN-γ and IL-6 than MPL or Poly I:C stimulation alone in PBMCs. In addition, the combination induced significantly higher mRNA expression of IL-1β, IL-6, and IL-12p40 in MoDCs, and IL-8, MCP-1, and VEGF in BM-MSCs compared to stimulation with a single TLR agonist. Conclusions: The combination of MPL and Poly I:C can be used as a potential adjuvant candidate for vaccines to aid in preventing infectious diseases in horses.

Rat Hindlimb Allotransplantation with Short-term Immune Suppressants and Dendritic Cell Pretreatment (단기간 면역억제제와 수지상 세포주의 전처치를 이용한 복합조직 동종이식)

  • Eun, Seok-Chan;Baek, Rong-Min
    • Archives of Reconstructive Microsurgery
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    • v.21 no.1
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    • pp.34-40
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    • 2012
  • Prevention of acute rejection in composite tissue allotransplantation without continuous immunosuppression lacks reports in worldwide literature. Recently dendritic cells (DC) gained considerble attention as antigen presenting cells that are also capable of immunologic tolerance induction. This study assesses the effect of alloantigen-pulsed dendritic cells in induction of survival in a rat hindlimb allograft. We performed hindlimb allotransplantation between donor Sprague-Dawley and recipient Fischer344 rats. Recipient derived dendritic cells were harvested from rat whole blood and cultured with anti-inflammatory cytokine IL-10. Then donor-specific alloantigen pulsed dendritic cells were reinjected into subcutaneous tissue before limb transplantation. Groups: I) untreated (n=6), II) DC injected (n=6), III) Immunosuppressant (FK-506, 2 mg/Kg) injected (n=6), IV) DC and immunouppressant injected (n=6). Graft appearance challenges were assessed postoperatively. Observation of graft appearance, H-E staning, immunohistochemical (IHC) study, and confocal immunofluoreiscece were performed postoperatively. Donor antigen pulsed host dendritic cell combined with short-term immunosuppression showed minimal mononuclear cell infiltration, regulator T cell presence, and could prolong limb allograft survival.

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Necrotizing meningoencephalitis in a pug dog in Korea (Pug dog에서 발생한 괴사성 뇌막뇌염 증례보고)

  • Bak, Eun-jung;Kim, Dae-yong;Kim, Jae-hoon;Kweon, Oh-hyeong
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.441-445
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    • 1996
  • A case of necrotizing meningoencephalitis was described in a 3-year-old female pug dog. Significant histopathological changes were confined to the brain. Histopathologically, brain lesion of the pug dog was characterized by (1) severe lymphocyte, plasma cell and macrophage infiltration in the meninges and gray and white matter of the cerebral hemisphere, (2) perivascular mononuclear cell cuffing and (3) focal polioencephalomalacia in the gray matter of cerebrum. Replicate sections of the brain and lung were negative for fungal infection, canine distemper and toxoplasmosis on PAS stain and immunohistochemistry, respectively. Based on clinical signs, histopathological and immunohistochemical studies, this case was diagnosed as necrotizing meningoencephalitis in pug dog that had not been preriously described in Korea.

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Effects of Ginseng Saponin on the Cytokine Gene Expression in Human Immune System (인삼 사포닌이 인간면역계 사이토카인 유전자의 발현에 미치는 영향)

  • 박종욱;한인숙
    • Journal of Ginseng Research
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    • v.20 no.1
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    • pp.15-22
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    • 1996
  • In order to investigate the Immunomodulatory effects of ginseng, we have studied the effects of ginseng saponin on the proliferation and cytosine gene expression of human pheripheral blood mononuclear cell (PBMC). In the PBMC proliferation assay, total saponin exhibited proliferation inhibition on the PBMC or phytohemagglutinin(PHA)-stimulated PBMC in a dose-dependent fashion. Immunomodulatory effects of ginseng were further investigated using the cytokine gene expression as the indicators. In the reverse transcription-polymerase chain reaction (RT-PCR) test, interleukin (IL)-1, IL-2, IL-3, IL-4, IL-6, IL-13, granulocyte macrophage-colony stimulating factor, tumor necrosis factor (TNF), migration inhibitory factor and transforming growth factor genes were expressed in the PHA-stimulated PBMC 48 hrs after cell culture. Among expressed cytokines, total saponin could increase the expression of IL-1 and TNF of PBMC without stimulation of PHA. All of ginsenosides, $Rb_1$, $Rb_2$, $Rg_1$, Rc, Re, incresed TNF gene expression. Especially, Rb2 (20 g/ml) showed most prominent effect on TNF gene expression and it also slightly increased IL-1 gene expression of PBMC.

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EFFECTS OF NITRIC OXIDE SYNTHASE INHIBITORS ON OSTEOCLAST-LIKE CELL FORMATION

  • Ahn, Seung-Kyu;Kim, Jung-Kun;Cha, Kyung-Suk
    • The korean journal of orthodontics
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    • v.25 no.6 s.53
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    • pp.715-722
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    • 1995
  • Orthodontic tooth movement in response to orthodontic force results from actions of osteoclasts and osteeoblasts in the cell level. Convincing evidence has now been provided to support the view that osteoclasts are derived from mononuclear cells that originate in the bone marrow or other hematopoietic organs and they migrate to the bones via vascular routes. Nitric oxide(NO), which accounts for the biological properties of endothelium-derived relaxing factor(EDRF), is the endogenous stimulator of soluble guanylate cylase. The discovery of the formation of nitric oxide(NO) from L-arginine in mammalian tissues and its biological roles has, in the last 7 years, thrown new light onto many areas of research. Data from experiments in vitro showed that N-metyl-L-arginine(L-NMA) and L-nitro-L- arginine(L-NAME) are competitive inhibitors of nitric oxide synthase. This study suggest that the multinucleated cells in our culture have characteristics of osteoclasts and that the potential bone cell activity of nitric oxide in vitro may be mediated in part by stimulation of marrow mononuclear cells to form osteoclast-like cells. Bone marrow cells were obtaineed from tibia of 19-days old chick embryo. After sacrifice, tibia was quickly dissected and the bone were then split to expose the medullary bone. The cells were attached for 4 hours and the nonadherent cells were collected. Marrow cells weere cultured in 96-well plate in medium 199. To examine the number of TRAP-positive multinucleated cells(MNCs), $10^{-8}\;M\;Vit=D_3$ and various concentration of L-NMA and L-NAME weere added at the beginning of cultures and with each medium change. After 7 days of culture. tartrate-resistant acid phosphatase(TRAP) staining was performed for microscopic evaluation. Cells haying more than three nuclei per cell were counted as MNCs. The obsrved results were as follows;1. 1,25-dihydroxyvitamine $D_3$ stimulated the osteoclast-like multinucleated cells in cultures of chick embryo bone marrow. 2. Nitric oxide synthase inhibitors(NOSI ; N-NMA, N-NAME) stimulated the osteoclast-like cells in cultures of chick embry bone marrow. 3. 1,25-dihydroxyvitamine$D_3$ and nitric oxide synthase inhibitors did not appear to have additive effect on the generation of TRAP-positive MNCs. These results suggest that nitric oxide synthase inhibitors may stimulate the osteoclast-like multinucleated cell formation and fusion in cultures of chick bone marrow.

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Evaluation for Cytopreservability of Manual Liquid-Based Cytology $Liqui-PREP^{TM}$ and its Application to Cerebrospinal Fluid Cytology: Comparative Study with Cytospin (수기 액상세포검사 $Liqui-PREP^{TM}$의 세포보존력 평가 및 뇌척수액 세포검사에의 적용: 세포원심분리법과의 비교)

  • Park, Gyeong-Sin;Lee, Kyung-Ji;Jung, Chan-Kwon;Lee, Dae-Hyoung;Cho, Bin;Lee, Youn-Soo;Shim, Sang-In;Lee, Kyo-Young;Kang, Chang-Suk
    • The Korean Journal of Cytopathology
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    • v.18 no.1
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    • pp.46-54
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    • 2007
  • Cerebrospinal fluid (CSF) cytology is an effective tool for evaluating diseases involving the central nervous system, but this technique is usually limited by its low cellularity and poor cellular preservation. Here we compared the manual liquid-base $Liqui-PREP^{TM}$ (LP) to the cytospin (CS) with using a mononuclear cell suspension and we applied both methods to the CSFs of pediatric leukemia patients. The cytopresevability, in terms of cell yield and cell size, and the clinical efficacy were evaluated. When 2000 and 4000 mononuclear cells were applied, LP was superior to CS for the cell yield, 16.8% vs 1.7% (P=0.001) and 26.2% vs 3.5% (P=0.002), respectively. The mean size of the smeared cells was 10.60 ${\mu}m$ in the CS, 5.01 ${\mu}m$ in the LP and 6.50 ${\mu}m$ in the direct smear (DS), and the size ratio was 1.7 (CS to DS), 0.8(LP to DS) and 2.1 (CS to LP), respectively. As compared to the cells in the DS, the cells in the CS were significantly enlarged, but those in the LP were slightly shrunken. Upon application to 109 CSF samples, 4 were diagnosed as positive for leukemia (positive), 4 had atypical cells and 101 were negative by CS; 6 were positive, one had atypical cells and 102 were negative by LP. For six cases, in which 4 were positive for leukemia and 2 of 4 had atypical cells by CS, they were positive by LP and they were also confirmed as positive according to the follow-up study. Three cases diagnosed as atypical cells (two by CS and one by LP), were confirmed as negative. In conclusion, these results suggest that LP is superior to CS for the cytopresevability and for rendering a definite diagnosis of cerebrospinal fluid.

A Study on Relationship of Symptom Distress and Natural Killer Cell Cytotoxicity in Breast Cancer Patients (치료 중인 유방암 환자의 신체적 증상과 자연살해세포 활성도의 관계)

  • Chae, Young-Ran
    • Journal of Korean Biological Nursing Science
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    • v.4 no.2
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    • pp.69-77
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    • 2002
  • The purpose of this study was to identify the relationship of symptom distress and natural killer cell cytotoxicity in breast cancer patients who had been radiation therapy and/or chemotherapy after surgery. Symptom distress measured by modified Lee's(1994) physical symptom questionnaire. For measuring the natural killer cell cytotoxic activity. 8ml to 10ml blood was collected from the subjects. Mononuclear cell was isolated by centrifuge of the blood and cultured by putting $Cr^{51}$, and reacted with target cell, K562 cell. Amount of $Cr^{51}$ was measured, and %lysis was calculated. The results were as follows. 1) Symptom distress score was 42.18, which is moderate symptom distress. 2) Natural killer cell cytotoxic activities were 42.18%lysis(effector : target cell ratio=100 : 1) and 28.05%lysis(effector : target cell ratio=50 : 1). 3) Correlation coefficients of symptom distress and natural killer cell cytotoxic activity were $-.134{\sim}-.461$. Though significant correlation was not found between total score of symptom distress and natural killer cell cytotoxic activity, 3('pain' 'feel hot on radiation site' and 'difficulty in breathing') of 19 symptom distress items and natural killer cell cytotoxic activity showed significant negative correlation(p<.05). These findings suggest that 1) breast cancer patients who had been radiation therapy and/or chemotherapy after surgery have moderate symptom distress and decreased natural killer cell cytotoxic activity. 2) The symptom distress was not related to natural killer cell cytotoxic activity.

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3C8, a new monoclonal antibody directed against a follicular dendritic cell line, HK

  • Lee, In Yong;Lee, Joonhee;Park, Weon Seo;Nam, Eui-Cheol;Shin, Yung Oh;Choe, Jongseon
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.26-31
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    • 2001
  • Background : Follicular dendritic cells (FDCs) play key roles during T cell-dependent humoral immune responses by allowing antigen-specific B cells to survive, proliferate, and differentiate within the FDC networks of secondary follicles, i.e., germinal centers (GC). Methods: A novel monoclonal antibody, 3C8, was generated by immunizing with an FDC line HK, in order to understand the molecular signals involved in the FDC-B cell interactions in the microenvironment of the GC. Results: The 3C8 antibody did not bind to mononuclear cells, including T cells, B cells, and monocytes. Murine L929 and human skin fibroblasts exhibited no or little reactivity to 3C8. However, 3C8 specifically recognized HK cells by flowcytometry. Furthermore, the antigen recognized by 3C8 was restricted to the GC of the human tonsil. Dendritic networks of the GC were intensely stained by 3C8, but cells outside the GC were not. Conclusion: Our results suggest that the antigen 3C8 may play some unique role on FDCs during the GC reactions.

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Fine Needle Aspiration Cytology of Ki-1 Positive Large Cell Lymphoma- A Case Report - (Ki-1양성 대세포림프종의 세침흡인 세포학적 소견 - 1예 보고 -)

  • Kwon, Kye-Hyun;Kwak, Jeong-Ja;Jin, So-Young;Lee, Dong-Wha
    • The Korean Journal of Cytopathology
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    • v.4 no.2
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    • pp.127-132
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    • 1993
  • Ki-1 lymphoma is a sort of high grade large cell lymphoma and defined on the basis of the reactivity of the tumor cells with monoclonal antibody Ki-1. On fine needle aspiration cytology, the reported case is rare and the differential diagnosis is not easy, especially from undifferentiated carcinoma and Hodgkin's lymphoma. We experienced a case of fine needle aspiration cytology of Ki-1 positive large cell lymphoma in a 61-year old male patient. Fine needle aspiration cytology from the cervical lymph node disclosed hypercellular smears with large single cells on polymorphous lymphoid background. The tumor cells had abundant dense cytoplasm and large nuclei with Irregular profiles. Although most cells were mononuclear binucleated and multilobed/multinucleated cells were also seen Immunohistochemistry was done and revealed strong positive staining for Ki-1 antigen.

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Intracranial Plasma Cell Granuloma

  • Kim, Dae-Jin;Choi, Yu-Seok;Song, Young-Jin;Kim, Ki-Uk
    • Journal of Korean Neurosurgical Society
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    • v.46 no.2
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    • pp.161-164
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    • 2009
  • Plasma cell granuloma is a tumor-like disease characterized by non-neoplastic polyclonal proliferation of plasma cells and other mononuclear cells. This disease occurs most frequently in the lung and upper respiratory tract, while the involvement of the central nervous system is very rare. A 44-year-old female patient presented with nausea and progressive visual disturbance. Brain magnetic resonance imaging (MRI) revealed the mass along the right tentorium with low signal intensity in the T2 weighted image (T2WI) and fluid-attenuated inversion recovery (FLAIR) sequence, and an isosignal intensity in T1 weighted image (T1WI), the latter of which was enhanced after administration of gadolinium-diethylenetriamine penta-acetic acid (Gd-DTPA). The thickest portion of the tentorium was partially excised via the combined suboccipital and infratentorial approach. The histopathological examination indicated a diagnosis of plasma cell granuloma. Postoperative steroid therapy was administered for remnant tumor control. Although a follow up MRI scan taken 20 months after the operation showed a slight decrease in tumor size, the lesion had extended to the falx and left frontal convexity along with parenchymal edema at 32 months after the operation and the clinical status was aggravated. The mass was removed from the left frontal convexity. Radiation therapy was given, together with steroid administration.