• Title/Summary/Keyword: Peripheral blood mononuclear cells(PBMCs)

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Inhibition of caspase-1-dependent apoptosis suppresses peste des petits ruminants virus replication

  • Lingxia Li;Shengqing Li;Shengyi Han;Pengfei Li;Guoyu Du;Jinyan Wu;Xiaoan Cao;Youjun Shang
    • Journal of Veterinary Science
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    • v.24 no.5
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    • pp.55.1-55.12
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    • 2023
  • Background: Peste des petits ruminants (PPR), caused by the PPR virus (PPRV), is an acute and fatal contagious disease that mainly infects goats, sheep, and other artiodactyls. Peripheral blood mononuclear cells (PBMCs) are considered the primary innate immune cells. Objectives: PBMCs derived from goats were infected with PPRV and analyzed to detect the relationship between PPRV replication and apoptosis or the inflammatory response. Methods: Quantitative real-time polymerase chain reaction was used to identify PPRV replication and cytokines expression. Flow cytometry was conducted to detect apoptosis and the differentiation of CD4+ and CD8+ T cells after PPRV infection. Results: PPRV stimulated the differentiation of CD4+ and CD8+ T cells. In addition, PPRV induced apoptosis in goat PBMCs. Furthermore, apoptosis and the inflammatory response induced by PPRV could be suppressed by Z-VAD-FMK and Z-YVAD-FMK, respectively. Moreover, the virus titer of PPRV was attenuated by inhibiting caspase-1-dependent apoptosis and inflammation. Conclusions: This study showed that apoptosis and the inflammatory response play an essential role in PPR viral replication in vitro, providing a new mechanism related to the cell host response.

Differential Expression of Th1- and Th2- Type Cytokines in Peripheral Blood Mononuclear Cells of Murrah Buffalo (Bubalus Bubalis) on TLR2 Induction by B. Subtilis Peptidoglycan

  • Shah, Syed M.;Ravi Kumar, G.V.P.P.S.;Brah, G.S.;Santra, Lakshman;Pawar, Hitesh
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.7
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    • pp.1021-1028
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    • 2012
  • Peripheral blood mononuclear cells (PBMCs) discriminate microbial pathogens and induce T-cell responses of appropriate effector phenotype accordingly. Toll-like receptors (TLRs), in part, mediate this microbial recognition and differentiation while the development of T-cell effector functions critically depends on the release of Th1- or Th2- type cytokines. In the present study, buffalo PBMCs were stimulated under in vitro culture conditions by Bacillus subtilis cell wall petidoglycan, a TLR2 ligand, in a dose- and time- dependent manner. The expression of TLR2 as well as the subsequent differential induction of the Th1 and Th2 type cytokines was measured. Stimulation was analyzed across five doses of peptidoglycan ($10{\mu}g/ml$, $20{\mu}g/ml$, $30{\mu}g/ml$, $40{\mu}g/ml$ and $50{\mu}g/ml$) for 3 h, 12 h, 24 h and 36 h incubation periods. We observed the induction of TLR2 expression in a dose- and time-dependent manner and the peptidoglycan induced tolerance beyond $30{\mu}g/ml$ dose at all incubation periods. The correlation between peptidoglycan stimulation and TLR2 induction was found positive at all doses and for all incubation periods. Increased production of all the cytokines was observed at low doses for 3 h incubation, but the expression of IL-4 was relatively higher than IL-12 at the higher antigen doses, indicating tailoring towards Th2 response. At 12 h incubation, there was a pronounced decrease in IL-4 and IL-10 expression relative to IL-12 in a dose- dependent manner, indicating skewing to Th1 polarization. The expression of IL-12 was highest for all doses across all the incubation intervals at 24 h incubation, indicating Th1 polarization. The relative expression of TNF-${\alpha}$ and IFN-${\gamma}$ was also higher while that of IL-4 and IL-10 showed a decrease. For 36 h incubation, at low doses, relative increase in the expression of IL-4 and IL-10 was observed which decreased at higher doses, as did the expression of all other cytokines. The exhaustion of cytokine production at 36 h indicated that PBMCs became refractory to further stimulation. It can be concluded from this study that the cytokine response to sPGN initially was of Th2 type which skews, more pronouncedly, to Th1 type with time till the cells become refractory to further stimulation.

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
    • Journal of Veterinary Clinics
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    • v.36 no.3
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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.

Banhabaikchulcheunma-tang Down-regulates LPS-induced Production of Pro-inflammatory Cytokines

  • Kim, Yun-Ha;Cho, Kwang-Ho;Moon, Byung-Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.1
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    • pp.192-198
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    • 2009
  • Banhabaikchulcheunma-tang (BBCT), a prescription composed of thirteen herbal mixtures, has been widely used in the treatment of brain disorders in Oriental Medicine. However, the mechanisms by which the formula affects on the production of pro-inflammatory cytokines in cerebral infarction (CI) patients remain unknown yet. The levels of secretory protein of pro-inflammatory cytokines, including tumor necrosis factor (TNF)-a, interlukin (IL)-1b, and IL-6, were significantly increased in lipopolysaccharide (LPS)-stimulated THP-1 differentiated macrophage-like (THP-1/M) cells and Peripheral blood mononuclear cells (PBMCs) from CI patients. However, pretreatment with BBCT significantly inhibited the secretion of pro-inflammatory cytokines, including TNF-a, IL-1b, and IL-6, in THP-1/M cells and PBMCs from CI patients with stimulus. Thus, these data indicate that BBCT may be beneficial in the cessation of inflammatory processes of cerebral infarction through suppression on the production of pro-inflammatory cytokines.

Cytokine-Inducing and T Cell Mitogenic Effects of Cordyceps hepialidicola

  • Lim, Jong-Soon;Kim, Seung-Hyung;Park, Jeong-Youl;Park, Jin-Seo;Park, Seong-Joo;Shin, Kwang-Soo
    • Journal of Microbiology
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    • v.39 no.3
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    • pp.181-185
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    • 2001
  • The morphological characteristics of newly isolated Cordyceps hepialidicola were characterized, and the phylogenetic relationships with other Cordyceps species were investigated using a sequence analysis of the internal transcribed spacer (ITS). The PCR product of 592 bp showed a homology of 92 and 91% with C. militaris and C. nutans, respectively, In an in vitro model using mouse peripheral blood mononuclear cells (PBMC), a methanol extract of C. hepialidicola induced multiple cytokines, including IFN-${\gamma}$ IL-4, and IL-18. The extract also enhanced the percentages of the CD4$\^$+/ and CD8$\sub$+/ T cells in the healthy murine PBMCs to 56.1% and 13.0%,respectively. The percentages of CD4$\^$+/ and CD8$\^$+/ in the untreated controls were 28.4 and 7.3%, and concanavalin A-treated positive controls were 62.4 and 18.3%, respectively.

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Genetic variation of BIV isolates characterized by PCR using degenerate primers

  • Kwon, Oh-Sik;Sninsky, John J.
    • Journal of Microbiology
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    • v.33 no.3
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    • pp.252-259
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    • 1995
  • The PCR was employed to detect and characterize the bovine immunodeficiency-like virus (BIV), which is a newly recognized member of the I entivirinae of the retroviruses. Degenerate primers representing the conserved regions in the pol genes of the Lentivirinae, were used to detect proviral DNA obtained from the bovine embryonic spleen cell cultures infected with BIV. The PCR amplified DNA fragment was molecularly cloned and sequenced. The BIV DNA fragment contained a sequence identical to that reported by Garvey et al. (Garvey et al., 1990. Virology, 175, 391-409). With the degenerate primers, peripheral blood mononuclear cells (PBMCs) of sick cattle and cells cultured with BIV were tested to determine genetic variation of BIV pol conserved sequence. We found the sequence heterogeneity within cultures and most variations occurred at the third base of codons that would not lead to amino acid substitutions. Another change was GAG (Glu) to AAG (Lys) within the BIV isolates. Interestingly, the altered sequence is also found in other lentiviruses such as HIV-2, SIV mac, CAEV and EIAV.

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Dopamine Receptor Gene (DRD1-DRD5) Expression Changes as Stress Factors Associated with Breast Cancer

  • Pornour, Majid;Ahangari, Ghasem;Hejazi, Seyed Hesam;Ahmadkhaniha, Hamid Reza;Akbari, Mohamad Esmail
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10339-10343
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    • 2015
  • Breast cancer is the most common cancer among females worldwide and a most prevalent malignancy in Iranian women. Chronic stress may make an important contribution to cancer, especially in the breast. Numerous studies showed roles of neurotransmitters in the occurrence and progression of cancers which are mediated by their various types of receptors. This study was conducted to evaluate alterations in the expression profile of dopamine receptor genes in peripheral blood mononuclear cells (PBMC) as stress factors in breast cancer patients and the human breast cancer cell line (MCF-7). Peripheral blood samples were obtained from 30 patients and 30 healthy individuals. Total mRNA was extracted from PBMC and MCF-7 cells and RT-PCR was performed to confirm the presence of five dopamine receptors (DRD1-DRD5). Expression changes of dopamine receptor genes were evaluated by real time PCR. We observed that DRD2-DRD4 in PBMCs of breast cancer patients were increased compared to healthy individuals. In addition, all dopamine receptor subtypes but DRD1 were expressed in MCF-7 cells. Therefore, alterations of these receptors as stress factors should be assessed for selecting appropriate drugs such as D2-like agonists for treatment of breast cancer after performing complimentary tests. Determining the expression profile of dopamine receptor genes thus seems promising.

Immunomodulatory activities of ethanolic extract of Drynariae Rhizoma (골쇄보(骨碎補) ethanol 추출물의 면역 조절 작용에 관한 연구)

  • Lee Ki-Uk;Jeong Ji-Cheon
    • The Journal of Internal Korean Medicine
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    • v.25 no.1
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    • pp.16-27
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    • 2004
  • In the traditional Chinese medicine, Drynariae Rhizoma (DR) has been reported as a good enhancer for bone healing. DR, a plant widely used in the traditional medicinal systems of Korea, has been reported to possess antiviral, antibacterial and anti-inflammatory activities. Modulation of immune response to alleviate disease has been of interest for a long time. Plant extracts have been widely investigated for possible immunomodulatory properties. Thus, I have evaluated the anticellular and immunomodulatory properties of ethanolic extract of DR. DR extract inhibited proliferation of mitogen (phytohaemagglutinin; PHA) and antigen (purified protein derivative; PPD)-stimulated human peripheral blood mononuclear cells (PBMCs). In addition, DR inhibited growth of several cell lines of mouse and human origin. It also inhibited production of nitric oxide (NO), interleukin-2 (IL-2) and tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$. Intracytoplasmic $interferon-{\gamma}\;(IFN-{\gamma})$ and expression of cell surface markers, CD16 and HLA-DR, on human PBMC, were not affected on treatment with DR but CD25 expression was down regulated. This study demonstrates the antiproliferative and immunosuppressive potential of ethanolic extract of DR in vitro.

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MicroRNA super-resolution imaging in blood for Alzheimer's disease

  • Mirae Lee;Jiwon Woo;Sang Tae Kim;Minho Moon;Sang Yun Kim;Hanna Cho;Sujin Kim;Han-Kyeol Kim;Jeong-Yoon Park
    • BMB Reports
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    • v.56 no.3
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    • pp.190-195
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    • 2023
  • We propose a novel blood biomarker detection method that uses miRNA super-resolution imaging to enable the early diagnosis of Alzheimer's disease (AD). Here, we report a single-molecule detection method for visualizing disease-specific miRNA in tissue from an AD mice model, and peripheral blood mononuclear cells (PBMCs) from AD patients. Using optimized Magnified Analysis of Proteome (MAPs), we confirmed that five miRNAs contribute to neurodegenerative disease in the brain hippocampi of 5XFAD and wild-type mice. We also assessed PBMCs isolated from the whole blood of AD patients and a healthy control group, and subsequently analyzed those samples using miRNA super-resolution imaging. We detected more miR-200a-3p expression in the cornu ammonis 1 and dentate gyrus regions of 3 month-old 5XFAD mice than in wild-type mice. Additionally, miRNA super-resolution imaging of blood provides AD diagnosis platform for studying miRNA regulation inside cells at the single molecule level. Our results present a potential liquid biopsy method that could improve the diagnosis of early stage AD and other diseases.

Effect on Immune Cells of Proteoglycan Originating from Rhanella aquatilis (Rhanella aquatilis 유래 당단백질의 면역세포에 미치는 영향)

  • Park, Hae-Gi;Kim, Kwang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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    • pp.312-315
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    • 2014
  • So as to evaluate its use an immune stimulator in humans, the toxicity and action against immune cells by an anti-yeast substance (AYS), a bacterial proteoglycan, were investigated. The AYS did not possess hemolytic activity with human red blood cells (hRBC). Nor did it exhibit cytotoxicity against human peripheral blood mononuclear cells (hPBMC). In addition, the AYS did not induce hPBMC proliferation, but it did agglutinate hPBMCs in vitro. Moreover, hPBMC induced inflammatory cytokines such as IL-6, IL-5, IFN-${\gamma}$ and TNF-${\alpha}$ with the AYS during culture. Compared with alum, the AYS as an adjuvant has an increased antibody production rate against bovine serum albumin (BSA) in mice.