• Title/Summary/Keyword: granulocyte colony-stimulating factor

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Effects of rrhGM-CSF on Morphology and Expression of PCNA in Regenerating Rat Liver (재생 중인 흰쥐 간의 형태학적 변화 및 PCNA 발현에 미치는 rrhGM-CSF의 영향)

  • Jeong, Jin-Ju;Heo, Si-Hyun;Kim, Ji-Hyun;Yoon, Kwang-Ho;Lee, Young-Jun;Han, Kyu-Boem;Kim, Wan-Jong
    • Applied Microscopy
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    • v.40 no.2
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    • pp.73-80
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    • 2010
  • Liver regeneration is a result of highly coordinated proliferation of hepatocytes and nonparenchymal liver cells. Partial hepatectomy (PH) is the most often used stimulus to study liver regeneration because, compared with other methods that use hepatic toxins, it is not associated with the tissue injury and inflammation, and the initiation of the regenerative stimulus is precisely defined. Granulocyte macrophage-colony stimulating factor (GM-CSF), which is a cytokine able to regulate the proliferation and differentiation of epithelial cells, was first identified as the most potent mitogen for bone marrow. Particularly, rrhGM-CSF, which is highly glycosylated and sustained longer than any other types of GM-CSF in the blood circulation, was specifically produced from rice cell culture. In this experiment, effects of rrhGM-CSF administration were evaluated in the regenerating liver after 78% PH of rats. Morphological changes induced by PH were characterized by destroyed hepatocyte plate around the central vein and enlarged nuclear cytoplasmic ratio and increased hepatocytes with two nuclei. And then, proliferation of liver cells (parenchymal and nonparenchymal) and rearrangement of plates and lobules seemed to be carried out during liver regeneration. These alterations in the experimental group preceded those of the control. Since proliferating cell nuclear antigen (PCNA) is known to be a nuclear protein maximally elevated in the S phase of proliferating cells, the protein was used as a marker of liver regeneration after PH in rats. PCNA levels by western blot analysis and immunohistology were compared between the two groups. PCNA protein expression of two groups at 12 hr and 24 hr after injury showed similar pattern. The protein expression showed the peak at 3 days in both groups, however, the protein level of the experimental group was higher than that of the control. On immunohistochemical observations, the reaction product of PCNA was localized at the nuclei of proliferating cells and the positive reaction in experimental group at 3 days was clearly stronger than that in control group. The results by Western blotting and immunohistology for PCNA showed similar pattern in terms of the protein levels. In conclusion, rrhGM-CSF administration during liver regeneration after 78% PH accelerated breakdown and restoration of the hepatic plate and expression of PCNA. These results suggest that rrhGM-CSF might play an important role during liver regeneration in rats.

EFFECT OF INTERLEUKIM-10 ON THE BONE RESORPTION INDUCED BY INTERLEUKIN-1B (Interleukin-10 이 $interleukin-1{\beta}$로 유도되는 골흡수에 미치는 효과)

  • Yu, Yun-Jung;Kang, Yun-Sun;Lee, Syng-Ill
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.321-339
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    • 1994
  • The cytokines released by osteoblasts induce bone resorption via the differentiation of osteoclast precursors. In this process, $interleukin-1{\beta}$($IL-1{\beta}$)-induced bone resorption is mediated by granulocyte macrophage-colony stimulation factor(GM-CSF), interleukin-6 (IL-6), and tumor necrosis factor ${\alpha}$($TNF-{\alpha}$) released from osteoblasts. Since these cytokines (GM-CSF, IL-6, $TNF-{\alpha}$) are produced by not only osteoblasts but also monocytes, and interleukin-10(I1-10) inhibits the secretion of these cytokines from monocytes, it may be speculated that IL 10 could modulate the production of GM-CSF, IL-6, and $TNF-{\alpha}$ by osteoblasts, then control $IL-1{\beta}-induced$ bone resorption. Therefore, the aims of the present study were to examine the effects of IL-10 on bone resorption. The sixten or seventeen-day pregnant ICR mice were injected with $^{45}Ca$ and sacrificed one day after injection. Then fetal mouse calvaria prelabeled with $^{45}Ca$ were dissected out. In order to confirm the degree of bone resorption, mouse calvaria were treated with Lipopolysaccharide(LPS), $TNF-{\alpha}$, $IL-1{\alpha}$, IL-8, $IL-1{\beta}$, and $IL-1{\alpha}$, Then, IL-10 and $interferon-{\gamma}$ ($IFN-{\gamma}$) were added to calvarial medium, in an attempt to evaluate the effect of $IL-1{\beta}-induced$ bone resorption. In addition, osteoclasts formation in bone marrow cell cultures, and the concentration of IL-6, $TNF-{\alpha}$, and GM-CSF produced from mouse calvarial cells were investigated in response to $IL-1{\beta}$ alone and simultaneously adding f $IL-1{\beta}$ and IL-10. The degree of bone resorption was expressed as the ratio of $^{45}Ca$ release(the treated/the control). The osteoclasts in bone marrow cultures were indentified by tartrate resistant acid phosphatase(TRAP) stain and the concentration of the cytokines was quantified using enzyme linked immunosorbent method. As results of these studies, bone resorption was induced by LPS(1 ng/ml ; the ratio of $^{45}Ca$ release, $1.14{\pm}0.07$). Also $IL-1{\beta}$(1 ng/ml), $IL-1{\alpha}$(1 ng/ml), and $TNF-{\alpha}$(1 ng/ml) resulted in bone resorption(the rations of $^{45}Ca$ release, $1.61{\pm}0.26$, $1.77{\pm}0.03$, $1.20{\pm}0.15$ respectively), but IL-8 did not(the ratio of $^{45}Ca$ release, $0.93{\pm}0.21$). The ratios of $^{45}Ca$ release in response to IL-10(400 ng/ml) and $IFN-{\gamma}$(100 ng/ml) were $1.24{\pm}0.12$ and $1.08{\pm}0.04$ respectively, hence these cytokines inhibited $IL-1{\beta}$(1 ng/ml)-induced bone resorption(the ratio of $^{45}Ca$ release $1.65{\pm}0.24$). While $IL-1{\beta}$(1 ng/ml) increased the number of TRAP positive multinulcleated cells in bone marrow cultures($20{\pm}11$), simultaneously adding $IL-1{\beta}$(1 ng/ml) and IL-10(400 ng/ml) decreased the number of these cells($2{\pm}2$). Nevertheless, IL-10(400 ng/ml) did not affect the IL-6, GM-CSF, and $TNF-{\alpha}$ secretion from $IL-1{\beta}$(1 ng/ml)-activated mouse calvarial cells. From the above results, it may be suggested that IL-10 inhibites $IL-1{\beta}-induced$ osteoclast differntiation and bone resorption. However, the inhibitory effect of IL-10 on the osteoclast formation seems to be mediated not by the reduction of IL-6, GM-CSF, and $TNF-{\alpha}$ production, but by other mechanisms.

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Effect of Recombinant Lactobacillus Expressing Canine GM-CSF on Immune Function in Dogs

  • Chung, Jin-Young;Sung, Eui-Jae;Cho, Chun-Gyu;Seo, Kyoung-Won;Lee, Jong-Soo;Bhang, Dong-Ha;Lee, Hee-Woo;Hwang, Cheol-Yong;Lee, Wan-Kyu;Youn, Hwa-Young;Kim, Chul-Joong
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1401-1407
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    • 2009
  • Many Lactobacillus strains have been promoted as good probiotics for the prevention and treatment of diseases. We engineered recombinant Lactobacillus casei, producing biologically active canine granulocyte macrophage colony stimulating factor (cGM-CSF), and investigated its possibility as a good probiotic agent for dogs. Expression of the cGM-CSF protein in the recombinant Lactobacillus was confirmed by SDS-PAGE and Western blotting methods. For the in vivo study, 18 Beagle puppies of 7 weeks of age were divided into three groups; the control group was fed only on a regular diet and the two treatment groups were fed on a diet supplemented with either $1\times10^9$ colony forming units (CFU)/day of L. casei or L. casei expressing cGM-CSF protein for 7 weeks. Body weight was measured, and fecal and blood samples were collected from the dogs during the experiment for the measurement of hematology, fecal immunoglobulin (Ig)A and IgG, circulating IgA and IgG, and canine corona virus (CCV)-specific IgG. There were no differences in body weights among the groups, but monocyte counts in hematology and serum IgA were higher in the group receiving L. casei expressing cGM-CSF than in the other two groups. After the administration of CCV vaccine, CCV-specific IgG in serum increased more in the group supplemented with L. casei expressing cGM-CSF than the other two groups. This study shows that a dietary L. casei expressing cGM-CSF enhances specific immune functions at both the mucosal and systemic levels in puppies.

Factors to Predict Successful Harvest during Autologous Peripheral Hematopoietic Stem Cell Collection

  • Kim, Mun-Ja;Jin, Soo-He;Lee, Duk-Hee;Park, Dae-Weon;Koh, Sung-Ae;Lee, Kyung-Hee;Hyun, Myung-Soo;Kim, Min-Kyoung
    • Biomedical Science Letters
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    • v.18 no.2
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    • pp.131-138
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    • 2012
  • Autologous peripheral blood stem cell transplantation (PBSCT) has been used as a major treatment strategy for hematological malignancies. The number of CD34 positive cells in the harvested product is a very important factor for achieving successful transplantation. We studied the factors that can predict the number of CD34 positive cells in the harvested product of acute myelocytic leukemia (AML), multiple myeloma (MM) and Non-Hodgkin's lymphoma (NHL) patients after mobilizing them with chemotherapy plus G-CSF. A total of 73 patients (AML 19 patients, MM 28 patients, NHL 26 patients) with hematological malignancies had been mobilized with chemotherapy and granulocyte colony-stimulating growth factor from April, 2000 to February, 2012. Group's characteristics, checkup opinion of pre-peripheral blood on the day of harvest & outcome of PBSC were analyzed and evaluated using SPSS statistics program after grouping patients as below; group 1: CD34 cell counts < $2{\times}10^6/kg$ (n=16); group 2: $2{\times}10^6/kg{\leq}CD34$ cell counts < $6{\times}10^6/kg$ (n=32); group 3: CD34 cell counts ${\geq}6{\times}10^6/kg$ (n=25). We analyzed the clinical characteristics, the peripheral blood (PB) parameters and the number of CD34 positive cells in the PB and their correlation with the yield of CD34 positive cells collected from the mobilized patients. The total number of leukapheresis sessions was 263 (mean: 3.55 session per patient), and the mean number of harvested CD34 positive cells per patient was $7.37{\times}10^6/kg$. The number of CD34 positive cells in product was significantly correlated with the number of platelet and CD34 positive cells in peripheral blood (P<0.05). The number of PB CD34 positive cells was the best significant factor for the quantity of harvested CD34 positive cells on the linear regression analysis (P<0.05). Many factors could influence the mobilization of peripheral blood stem cells. Platelet count and PB CD34 positive cells count were the two variables which remained to be significant in multivariate analysis. Therefore, the number of platelet and CD34 positive cells in peripheral blood on the day of harvest can be used as an accurate predictor for successful peripheral blood stem cell collection.

Effect of GM-CSF on Porcine Parthenotes Development (GM-CSF가 돼지 처녀 생식 배아 발달에 미치는 영향)

  • Lee, Jae-Dal
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.1
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    • pp.365-370
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    • 2015
  • Granulocyte-macrophage colony-stimulating factor (GM-CSF) is an important hematopoietic growth factor and immune modulator. The aim of this study was to evaluate the effects of GM-CSF on the development and cell number of porcine parthenotes, as well as on their expression of implantation-related genes. In the present study, porcine parthenogenatic activated embryos were cultured in a protein-free culture medium in the absence or presence of 5, 10 and 20 ng/ml GM-CSF for 7 days. The percentage of blastocyst formation, total cell number and gene expressions were evaluated. The results showed that the addition of 20 ng/ml GM-CSF to protein-free culture medium significantly increased the blastocoel formation ($26.14{\pm}2.03%$ vs. $3.55{\pm}0.51%$, p < 0.05). In addition, the cell number also increased when they were cultured in the presence of 20 ng/ml GM-CSF ($43.51{\pm}3.6%$ vs. $30.68{\pm}5.51%$, p < 0.05). A real time reverse transcripts polymerase chain reaction (RT-PCR) showed that GM-CSF enhances mRNA expression of the interleukin-6, but does not influence the leukemia inhibitory factor (LIF) receptor mRNA expression in blastocyst stage parthenotes. These results suggest that GM-CSF may enhance the viability of porcine embryos developing in vitro in a defined culture medium.

Inhibitory effect of Fagopyrum esculentum on degranulation and production of cytokine in RBL-2H3 cells (교맥의 RBL-2H3 비만세포 탈과립과 cytokine 생산 억제 효과)

  • Kang, Kyung-Hwa;Lee, Seung-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.25 no.3
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    • pp.1-12
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    • 2012
  • Objectives : Fagopyrum esculentum(FE) has been used for removal of inflammation of internal organs and treatment of sore and ulcer by heat toxin in Korean herbal medicines. In this study, To investigated the protective effect of FE on allergic response, we determined whether FE inhibits allergic response. Methods : The effect of FE was analyzed by ELISA, RT-PCR and Western blot in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase, as a marker of degranulation, cytokne, and intracellular ROS and MAPK and NF-${\kappa}B$ signaling. Results : We found that FE suppressed ${\beta}$-hexosaminidase release, the production of IL-4 and TNF-${\alpha}$ and intracellular ROS level in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. FE also significantly inhibited cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, p38 and $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ signal transduction pathway. Conclusions : Our results indicate that FE protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and ROS via the suppression MAPK and NF-${\kappa}B$ of signal transduction. Abbrevations : FE, Fagopyrum esculentum; RBL-2H3, rat basophilic leukemia cell line; ROS, reactive oxygen species; MAPK, Mitogen-activated protein kinase; $NF{\kappa}B$, nuclear factor ${\kappa}B$; $TNF{\alpha}$, Tumor necrosis factor alpha; GM-CSF, Granulocyte macrophage colony-stimulating factor; ERK, extracellular-signal-regulated kinase; JNK, c-Jun NH2-terminal kinase; p38, p38 MAP kinase; $I{\kappa}B{\alpha}$, inhibitory-kappa B alpha.

Rice Cell Origin Recombinant Human Granulocyte Macrophage Colony-Stimulating Factor (rrhGM-CSF) Could Improve the Wound Healing in Diabetic Hamster (당뇨가 유발된 햄스터 창상치유에 미치는 벼세포 유래 GM-CSF의 효과)

  • Han, Kyu-Boem;Heo, Si-Hyun;Jeong, Jin-Ju;Han, Man-Deuk;Kim, Wan-Jong;Shin, Kil-Sang
    • Applied Microscopy
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    • v.39 no.3
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    • pp.253-260
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    • 2009
  • GM-CSF is a multipotent growth factor, which also plays an important role during the process of wound healing. rrhGM-CSF was specifically produced from rice cell culture in our laboratory (Hanson Biotech Co., Ltd, Daejeon). The rrhGMCSF contains more oligosaccharide side chains than any other types of GM-CSF. This work was taken to evaluate the influence on wound healing of rrhGM-CSF in male golden hamsters. Full thickness skin defects of 9 mm in diameter were made in the back of hamsters, and 100 ${\mu}L$ ointment containing rrhGM-CSF 50 ${\mu}g/mL$ was applied. Control groups were given ointment without rrhGM-CSF. The wound sizes were relatively reduced and skin was well regenerated in the experimental group compared with the control group. Structurally, reepithelialization and architecture of the skin following injury were well accomplished in the experimental group. And also, positive reaction of PCNA of the skin following injury was more prominent in rrhGM-CSF containing ointment treatment group. Since this type of GM-CSF has highly glycosylated side chains, the effectiveness might be retain longer and stable, regarding acceleration of wound healing in the animal model. The present study has important implications for further development of the therapeutic manipulation of wound healing using rrhGM-CSF.

Effect of Angelicae Gigantis Radix for Inflammatory Response in HaCaT Cells (당귀(當歸) 추출물이 피부 각질형성세포의 염증반응에 미치는 영향)

  • Huh, Jung;Park, Hoyeon;Kim, Eom Ji;Kim, Eun-Young;Sohn, Youngjoo;Jung, Hyuk-Sang
    • The Korea Journal of Herbology
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    • v.37 no.3
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    • pp.9-19
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    • 2022
  • Objectives : Angelicae Gigantis Radix (AG) is a plant of the Ranunculus family. AG have been reported to have various pharmacological effects on human health which include uterine growth promotion, anti-inflammatory, analgesic, and immune enhancement. However, research on dermatitis disease is insufficient. Therefore, we investigated the effects of AG on tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ) stimulated HaCaT cell. Methods : To investigate the effect of AG on HaCaT cell, HaCaT cells were pre-treated with AG for 1 hour and then stimulated with TNF-α/IFN-γ. After 24 hours, media and cells were harvested to analyze the inflammatory mediators. Concentration of human interleukin-1beta (IL-1β), monocyte chemoattractant protein-1 (MCP-1), granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-α in the media were assessed by ELISA. mRNA expression of human thymus and activation-regulated chemokine (TARC), IL-6, and IL-8 were analyzed by RT-PCR. Additionally, the mechanisms of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway were investigated by Western blot. Results : The treatment of AG inhibited gene expression levels of IL-6, IL-8, and TARC and protein expression levels of IL-1β, MCP-1, and GM-CSF. Also, AG significantly reduced extracellular signal-regulated kinase (ERK) phosphorylation and NF-κB translocation in TNF-α/IFN-γ stimulated HaCaT cell. Conclusions : Taken together, these results demonstrate that AG can alleviate inflammatory diseases such as atopic dermatitis by regulating the expression of inflammatory cytokines. Also, it suggest that AG may a promising candidate drug for the treatment of inflammatory disease such as atopic dermatitis.

Chemical Characteristics and Immunostimulating Activity of Crude Polysaccharide Isolated from Commercial Instant Coffee (시판 인스턴트 커피에서 추출한 다당류의 화학적 특성 및 면역활성)

  • Kwak, Bong-Shin;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.48 no.3
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    • pp.289-295
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    • 2016
  • To elucidate the new biologically active ingredient in commercial instant coffee, a crude polysaccharide (ICP-0) was isolated by ethanol precipitation, and its immunostimulatory activity was estimated. ICP-0 mainly consisted of galactose (55.5%), mannose (25.7%), arabinose (6.0%), and galacturonic acid (10.1%), suggesting the possibility of its existence as a mixture of galactomannan or pectic polysaccharide. ICP-0 showed proliferative activity in peritoneal macrophages and splenocytes. ICP-0 dose-dependently augmented the production of nitric oxide and reactive oxygen species by peritoneal macrophages. In addition, murine peritoneal macrophages stimulated by ICP-0 showed enhanced production of various cytokines (tumor necrosis factor-${\alpha}$, interleukin-6, and interleukin-12) as compared to unstimulated murine peritoneal macrophages. In an in vitro assay for assessing intestinal immunomodulation, the ICP-0-treated Peyer's patch cells showed higher bone marrow cell proliferation activity at $100{\mu}g/mL$ and higher production of granulocyte-macrophage colony-stimulating factor, compared to the untreated Peyer's patch cells. These results suggest that polysaccharides in commercial instant coffee have a potentiality for macrophage functions and the intestinal immune system.

A Study on the Biological Activity of Allium cepa Extract in Vivo (양파 추출물의 in vivo 생리활성에 관한 연구)

  • Lee, Jin Young
    • Journal of Life Science
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    • v.30 no.3
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    • pp.267-277
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    • 2020
  • This study was conducted to assess the effect of highly concentrated onion intake on rodents. The experimental animals were divided two groups as follows; water administered group (CON) and Allium cepa administered group (ACE). The ACE group showed a slightly increases in the number of erythrocytes (RBC), hemoglobin (HGB), hematocrit (Hct) levels compared to the control group (p<0.05). Hemoglobin, monocyte, lymphocyte and neutrophil had no significant change (p>0.05) in ACE group and control group. The analysis of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) levels showed a significant decrease in ACE group compared to the control group (p<0.05). The blood glucose, total protein, HDL-cholesterol were slightly high in ACE group, while triglyceride, total cholesterol levels were lower in ACE group compared to the control group (p<0.05). The levels of cytokines (interluekin-1α (IL-1α), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), interferon-γ (INF-γ), interleukin-18 (IL-18), interleukin-2 (IL-2), granulocyte-macrophage colony-stimulating factor (GM-CSF)) involved in immunity and inflammation in liver tissue and blood have all been confirmed to be within normal range. These findings could be used as basic data to show that highly concentrated dietary onion extract is not toxic to hematological indicators and immune functions.