• Title/Summary/Keyword: PBMC

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Cell Cycle Regulation and Induction of Apoptosis by β-carotene in U937 and HL-60 Leukemia Cells

  • Upadhyaya, K.R.;Radha, K.S.;Madhyastha, H.K.
    • BMB Reports
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    • 제40권6호
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    • pp.1009-1015
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    • 2007
  • In this communication, we report the efficacy of $\beta$-carotene towards differentiation and apoptosis of leukemia cells. Dose ($20{\mu}M$) and time dependence (12 h) tests of $\beta$-carotene showed a higher magnitude of decrease (significance p < 0.05) in cell numbers and cell viability in HL-60 cells than U937 cells but not normal cell like Peripheral blood mononuclear cell (PBMC). Microscopical observation of $\beta$-carotene treated cells showed a distinct pattern of morphological abnormalities with inclusion of apoptotic bodies in both leukemia cell lines. When cells were treated with $20{\mu}M$ of $\beta$-carotene, total genomic DNA showed a fragmentation pattern and this pattern was clear in HL-60 than U937 cells. Both the cell lines, on treatment with $\beta$-carotene, showed a clear shift in $G_1$ phase of the cell cycle. In addition the study also revealed anti-oxidant properties of $\beta$-carotene since there was reduction in relative fluorescent when treated than the control at lower concentration. Collectively this study shows the dual phenomenon of apoptosis and differentiation of leukemia cells on treatment with $\beta$-carotene.

Characterization and Expression Profile of CMTM3/CKLFSF3

  • Zhong, Ji;Wang, Yu;Qiu, Xiaoyan;Mo, Xiaoning;Liu, Yanan;Li, Ting;Song, Quansheng;Ma, Dalong;Han, Wenling
    • BMB Reports
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    • 제39권5호
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    • pp.537-545
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    • 2006
  • CMTM/CKLFSF is a novel family of proteins linking chemokines and TM4SF. In humans, these proteins are encoded by nine genes, CKLF and CMTM1-8/CKLFSF1-8. Here we report the characteristics and expression profile of CMTM3/CKLFSF3. Human CMTM3/CKLFSF3 has a high sequence identity among various species and similar characteristics as its mouse and rat homologues. Established by results both of RT-PCR and Quantitative Real-time PCR, the gene is highly transcribed in testis, leukocytes and spleen. For further verification, we generated a polyclonal antibody against human CMTM3/CKLFSF3 and found that the protein is highly expressed in the testis and some cells of PBMCs. Therefore, CMTM3/CKLFSF3 is an evolutionarily conserved gene that may have important roles in the male reproductive system and immune system. Further studies are necessary to validate its functions in the two systems.

Characterization of Biocompatible Polyelectrolyte Complex Multilayer of Hyaluronic Acid and Poly-L-Lysine

  • Hahn, Sei-Kwang;Allan S. Hoffman
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권3호
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    • pp.179-183
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    • 2004
  • A biocompatible polyelectrolyte complex multilayer (PECML) film consisting of poly-L-lysine (PLL) as a polycation and hyaluronic acid (HA) as a polyanion was developed to test its use for surface modification to prevent cell attachment and protein drug delivery. The formation of PECML through the electrostatic interaction of HA and PLL was confirmed by contact angle measurement, ESCA analysis, and HA content analysis. HA content increased rapidly up to 8 cycles for HA/PLL deposition and then slightly increased with an increasing number of deposition cycle. In vitro release of PLL in the PECML continued up to 4 days and ca. 25% of HA remained on the chitosan-coated cover glass after in vitro release test for 7 days. From the results, PECML of HA and PLL appeared to be stable for about 4 days. The surface modification of the chitosan-coated cover glass with PECML resulted in drastically reduced peripheral blood mononuclear cell (PBMC) attachment. Concerned with its use for protein drug delivery, we confirmed that bovine serum albumin (BSA) as a model protein could be incorporated into the PECML and its release might be triggered by the degradation of HA with hyaluronidase.

Construction of Novel Bifunctional Chimeric Proteins Possessing Antitumor and Thrombolytic Activities

  • Hui, Jing;Dai, Youjin;Bian, Yuanyuan;Li, Hui;Cui, Xiaojin;Yu, Xiaojie;You, Song;Hu, Fengqing
    • Journal of Microbiology and Biotechnology
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    • 제22권7호
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    • pp.894-901
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    • 2012
  • Based on their respective antitumor and thrombolytic activities, the superantigen staphylococcal enterotoxin C2 (SEC2) and staphylokinase (Sak) were chosen for the construction of the novel chimeric proteins Sak-linker-SEC2 and SEC2-linker-Sak using a linker composed of nine Ala residues. Both chimeric proteins possessed nearly the same PBMC proliferation stimulating activity and antitumor activity as SEC2 and thrombolytic activity as Sak. Neither the SEC2 or Sak component of each chimeric protein affected the activity of the other component. The results presented in this study provide a possible strategy to prevent and cure tumor thrombus.

Streptozotocin-Induced Pro-Inflammatory Response in Rat Blood and its Attenuation by External Albumin Administration

  • Bae, Chun-Sik;Lee, Chang-Min;Kim, Dong-il;Ahn, Taeho
    • 한국임상수의학회지
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    • 제36권3호
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    • pp.145-149
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    • 2019
  • Previous report has suggested that the albumin levels were reduced in the peripheral blood mononuclear cells (PBMCs) and consequently oxidative stress was elevated in streptozotocin (STZ)-induced diabetic rats as albumin is the predominant antioxidant in plasma. In this study, we suggest that the levels of pro-inflammatory cytokine such as interleukin-6 (IL-6) and tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) were increased by approximately 3.4- and 2.9-fold, respectively, in the serum of STZ-diabetic rats, compared to those of normal rat. In addition to the cytokines, the levels of C-reactive protein (CRP) were also about 3.6-fold higher, indicating that STZ induced a pro-inflammatory response in rat blood. However, when purified rat albumin was externally co-administrated with STZ through the tail vein, the serum levels of IL-6, $TNF-{\alpha}$, and CRP were markedly reduced, although the values were still higher than those of normal (non-diabetic) rats. Albumin administration also decreased STZ-induced oxidative stress in serum and PBMCs. Moreover, the decrease in cytokine and CRP levels was dependent on the dose of injected albumin. These results suggest that STZ-induced pro-inflammation and oxidative stress in rat blood might be attenuated by treatment with exogenous albumin.

Composition for the Immunity Stimulatory Activity Comprising Calystegia dahuricus (Herb.) Choisy Water Extract

  • Jeong, Myeongguk;Kwon, Hyeokjin;Jeong, Seohye;Seo, Yerin;Kim, Minguk;Choi, Go-Eun
    • 대한의생명과학회지
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    • 제28권3호
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    • pp.206-210
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    • 2022
  • Calystegia dahurica (Herb.) Choisy is a natural product that has not been studied for efficacy or active ingredients. The purpose of this study is to investigate the activation effect of natural killer cells using a natural extract composition based on Calystegia dahurica (Herb.) Choisy extract (CDCE). We evaluated the activity of natural killer cells in natural products using PBMCs from healthy participants. All natural products were extracted with 50% ethanol. Based on the results of the cell viability assay, PBMCs of healthy participants were treated with extracts at various concentrations. Then, analysis was performed using flow cytometry to measure the cd107a surface expression of natural killer cells. As a result, treatment with a single extract of PBMCs increased the expression of cd107a in a concentration-dependent. Furthermore, it was confirmed that the treatment of the extract composition showed the highest expression of cd107a. In conclusion, it is expected that the extract composition containing CDCE according to this study can be used for prevention or treatment of cancer cells, tumor cells, and immune diseases.

Effect of Dietary Supplementation of Procyanidin on Growth Performance and Immune Response in Pigs

  • Park, J.C.;Lee, S.H.;Hong, J.K.;Cho, J.H.;Kim, I.H.;Park, S.K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권1호
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    • pp.131-139
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    • 2014
  • This study was performed to determine the effect of dietary supplementation of procyanidin on growth performance, blood characteristics, and immune function in growing pigs. In experiment 1 (Exp. 1), thirty-two crossbred pigs with an initial BW of $19.2{\pm}0.3$ kg were allocated into 4 treatments for an 8-wk experiment: i) CON (basal diet), ii) MOS 0.1 (basal diet+0.1% mannanoligosaccharide), iii) Pro-1 (basal diet+0.01% procyanidin), and iv) Pro-2 (basal diet+0.02% procyanidin). Pigs fed Pro-1 and Pro-2 diets had greater (p<0.05) gain:feed ratio compared with those fed CON or MOS 0.1 diets. Serum creatinine concentration was less (p<0.05) in Pro-2 treatment than those in CON, MOS 0.1 and Pro-1 treatments. In Exp. 2, twelve pigs (BW $13.4{\pm}1.3$ kg) received basal diet with i) 0 (CON), ii) 0.02% (Pro-0.02%), and iii) 0.04% procyanidin (Pro-0.04%) for 4 wk. Concentration of platelets was lower (p<0.05) in the Pro-0.04% group compared to CON at 24 h after lipopolysaccharide (LPS) challenge. In addition, secretion of cytokines from cultured peripheral blood mononuclear cells (PBMC) in the presence or absence of procyanidin was examined. The levels of interleukin (IL)-$1{\beta}$, IL-6 and tumor necrosis factor (TNF)-${\alpha}$ were lower (p<0.05) in Pro (LPS-stimulated PBMCs+procyanidin) than those in CON (LPS-stimulated PBMCs+PBS) at 4 h after LPS challenge. These data suggest that dietary addition of procyanidin improves feed efficiency and anti-inflammatory cytokines of pigs.

배독환(排毒丸)의 아토피피부염 병태모델에서의 면역 억제 효능 (Effects of Baedokhwan on Immune Modulation in Atopic Dermatitis Model of NC/Nga Mice)

  • 이종협;김선빈;김동희
    • 동의생리병리학회지
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    • 제24권5호
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    • pp.796-806
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    • 2010
  • Atopic dermatitis induced NC/Nga mice were used to investigate the efficacy of BDH(Baedokhwan) on the recovery of dermatitic symptoms through its influence on the immune related factors and histological changes. First of all, BDH treated group showed improvement of atopic dermatitis with naked eye observation, and significant decrease of clinical index(CI) was observed after 14 weeks. And Infiltration of leukocytes was suppressed in BDH treated group, and the thickness of hypertrophied epidermis and dermis were decreased. In dorsal skin, BDH treated group showed significant decrease of the ratio of CD3+, CD11b+/Gr-1+ immune cells by 52.8%, 25.2, respectively. And also significant decrease the level of IL-5 mRNA and IL-13 mRNA by 44.4%, 28.0, respectively. In PBMC and serum, BDH treated group showed an decrease of CD4+/CD45+, B220+/CD23+, CD4+, CD8+, CD4+/CD25+ immune cells by 35.0%, 12.6%, 42.7%, 31.6% and 55.6%, respectively, and the level of histamine was decreased by 39.0%. The results above indicated that BDH clinically used for atopic dermatitis treatment has objective validity, and therefore can be provided as the basic data for anti-allergic and anti-inflammatory studies.

Collagen II로 유도된 CIA 관절염 생쥐의 대한 가미해동피산(加味海桐皮散)의 억제 효과 (Suppressive Effects of GHS in Knee Joint, Regional Lymph Nodes, and Spleen on Collagen-induced Arthritis in Mice)

  • 김남욱;김동희
    • 동의생리병리학회지
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    • 제21권5호
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    • pp.1233-1242
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    • 2007
  • Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which might be mediated by the altered activation of Immune system, ultimately leading to the destruction of cartilage and bone. To examine effects of GHS on rheumatoid arthritis DBA/1J mice were immunized with bovine type II collagen to induced arthritis and then treated with GHS once a day for 7 weeks. Oral administration of GHS (200 mg/Kg) significantly suppressed the progression of CIA, which extend is comparable to that of methotrexate (MTX, 0.3 mg/Kg), a positive control. The severity of arthritis within the knee joints, which was evaluated by histological assessment of cartilage destruction and pannus formation, was also lowered by GHS. The production of TNF-and IL-6 in serum was significantly suppressed. The levels of IFN-g in the culture supernatant of splenocytes stimulated with CD3/CD28 or collagen were dramatically decreased, while those of IL-4 was increased. The levels of IgG and IgM RA factor were also decreased in the serum. FACS analysis indicated that B cells (in DLN), CD3+ T cells (in spleen, and paw joint), CD11b+Gr-1+ cells (in paw joint), CD3+CD49b(DX5) (in PBMC) were decreased and there was increased proportion of CD3+, CD4+, CD8+, CD4+CD25+ T cells in DLN. In conclusion, our results demonstrates that GHS significantly suppressed the progression of CIA and this action was characterized by the decreased production of TNF-a, IL-6, and rheumatoid factors, and modulations of immune cell populations.

아토피피부염 동물 병태모델에서의 형개연교탕(荊芥連翹湯)의 면역조절작용 (The effects of HYT on various immunological factors related to pathogenesis of allergic dermatitis in NC/Nga mice induced by Biostir AD)

  • 강란이;박보경;김선빈;최학주;김동희
    • 혜화의학회지
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    • 제18권2호
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    • pp.47-62
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    • 2009
  • Atopic dermatitis induced NC/Nga mice were used to investigate the efficacy of HYT on the recovery of dermatitic symptoms by how HYT influenced the immune related factors. The results are as below: 1. Compared to the control, HYT treated group showed recovery of atopic dermatitis by the naked eye observation, and significant reduction of dermatits index was observed after 14 weeks. 2. HYT treated group showed significant decrease of the ratio of CCR3+, B220+/IgE+, and Gr-1+/CD11b+ immune cells in dorsal skin by 40.5%, 34.2%, and 48.1%, respectively. 3. HYT treated group showed increase in the ratio of CD19+ immune cells within PBMC by 10.8%, as well as decrease in CD3+, CD3+/CD69+, NKT+ ratios by 5.3%, 35.2%, and 44.9%, respectively. 4. HYT treated group showed increase in the expression of IFN-$\gamma$ in serum by 589.3%, whereas the expression of IL-4, IL-5, IL-6, IL-13, TNF-$\alpha$, MCP-1 and RANTES were decreased by 31.4%, 82.1%, 97.1%, 39.5%, 83.7%, 26.1%, 48.6%, respectively. A 47.2% decrease in IgE expression was also observed. The results above strongly supported the improvement of atopic dermatitis by HYT treatment through immune modulation. Further studies on the synergistic effect of each ingredients of HYT and therapeutic effects according to the dosage of each ingredient should be followed for clinical applications. This work was (partly) supported by the RIC program of MKE(Ministry of Knowledge Economy) in Daejeon University.

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