• Title/Summary/Keyword: immune marker

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Influence of Toxoplasma gondii Acute Infection on Cholinesterase Activities of Wistar Rats

  • Tonin, Alexandre Alberto;da Silva, Aleksandro Schafer;Thorstenberg, Maria Luiza;Castilhos, Livia Gelain;Franca, Raqueli Teresinha;Leal, Daniela Bitencourt Rosa;Duarte, Marta Maria Medeiros Frescura;Vogel, Fernanda Silveira Flores;de La Rue, Mario L.;dos Anjos Lopes, Sonia Terezinha
    • Parasites, Hosts and Diseases
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    • v.51 no.4
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    • pp.421-426
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    • 2013
  • Several studies have shown the mechanisms and importance of immune responses against Toxoplasma gondii infection and the notable role of cholinesterases in inflammatory reactions. However, the association between those factors has not yet been investigated. Therefore, the aim of this study was to evaluate the acetylcholinesterase (AChE) activity in blood and lymphocytes and the activity of butyrylcholinesterase (BChE) in serum of rats experimentally infected with T. gondii during the acute phase of infection. For that, an in vivo study was performed with evaluations of AChE and BChE activities on days 5 and 10 post-infection (PI). The activity of AChE in blood was increased on day 5 PI, while in lymphocytes its activity was enhanced on days 5 and 10 PI (P<0.05). No significant difference was observed between groups regarding to the activity of BChE in serum. A positive (P<0.01) correlation was observed between AChE activity and number of lymphocytes. The role of AChE as an inflammatory marker is well known in different pathologies; thus, our results lead to the hypothesis that AChE has an important role in modulation of early immune responses against T. gondii infection.

Rapid, Sensitive, and Specific Detection of Salmonella Enteritidis in Contaminated Dairy Foods using Quantum Dot Biolabeling Coupled with Immunomagnetic Separation

  • Kim, Hong-Seok;Chon, Jung-Whan;Kim, Hyunsook;Kim, Dong-Hyeon;Yim, Jin-Hyuk;Song, Kwang-Young;Kang, Il-Byung;Kim, Young-Ji;Lee, Soo-Kyung;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.4
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    • pp.271-275
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    • 2015
  • Colloidal semiconductor CdSe-ZnS core-shell nanocrystal quantum dot (Qdot) are luminescent inorganic fluorophores that show potential to overcome some of the functional limitations encountered with organic dyes in fluorescence labeling applications. Salmonella Enteritidis has emerged as a major cause of human salmonellosis worldwide since the 1980s. A rapid, specific, and sensitive method for the detection of Salmonella Enteritidis was developed using Qdot as a fluorescence marker coupled with immunomagnetic separation. Magnetic beads coated with anti-Salmonella Enteritidis antibodies were employed to selectively capture the target bacteria, and biotin-conjugated anti-Salmonella antibodies were added to form sandwich immune complexes. After magnetic separation, the immune complexes were labeled with Qdot via biotin-streptavidin conjugation, and fluorescence measurement was carried out using a fluorescence measurement system. The detection limit of the Qdot method was a Salmonella Enteritidis concentration of $10^3$ colony-forming units (CFU)/mL, whereas the conventional fluorescein isothiocyanate-based method required over $10^5CFU/mL$. The total detection time was within 2 h. In addition to the potential for general nanotechnology development, these results suggest a new rapid detection method of various pathogenic bacteria from a complex food matrix.

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Immunomodulatory activities of polysaccharides extracted from Cudrania tricuspidata fruits in macrophage (꾸지뽕(Cudrania tricuspidata) 열매에서 분리된 조다당의 큰포식세포 면역 활성 조절)

  • Cho, Eun-Ji;Kim, Yi-Eun;Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.50 no.5
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    • pp.511-516
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    • 2018
  • Macrophages play a crucial role in the host immune defense system. The current study investigated immunomodulatory activities induced by polysaccharides extracted from Cudrania tricuspidata (CTPS) fruits in murine macrophages and their role in signaling pathways. In macrophages, CTPS predominantly induced nitric oxide (NO), tumor necrosis factor-a, and interleukin-6 production. In addition, CTPS significantly up-regulated expression of the macrophage surface marker (CD80/86 and MHC class I/II). These results indicate that polysaccharides extracted from CTPS may potentially play an immunomodulatory role in macrophages via mitogen-activated protein kinases and nuclear factor-B signaling. These findings may be useful in the development of immune enhancing adjuvant materials obtained from natural sources.

Value of Combined Detection of Serum CEA, CA72-4, CA19-9 and TSGF in the Diagnosis of Gastric Cancer

  • Yin, Li-Kui;Sun, Xue-Qing;Mou, Dong-Zhen
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3867-3870
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    • 2015
  • Background: To explore whether combined detection of serum tumor markers (CEA, CA72-4, CA19-9 and TSGF) improve the sensitivity and accuracy in the diagnosis of gastric cancer (GC). Materials and Methods: An automatic chemiluminescence immune analyzer with matched kits were used to determine the levels of serum CEA, CA72-4, CA19-9 and TSGF in 45 patients with gastric cancer (GC group), 40 patients with gastric benign diseases (GBD group) hospitalized in the same period and 30 healthy people undergoing a physical examination. The values of those 4 tumor markers in the diagnosis of gastric cancer was analyzed. Results: The levels of serum CEA, CA72-4, CA19-9 and TSGF of the GC group were higher than those of the GBD group and healthy examined people and the differences were significant (P<0.001). The area under receiver operating characteristic (ROC) curves for single detection of CEA, CA72-4, CA19-9 and TSGF in the diagnosis of GC was 0.833, 0.805, 0.810 and 0.839, respectively. The optimal cutoff values for these 4 indices were 2.36 ng/mL, 3.06 U/mL, 5.72 U/mL and 60.7 U/mL, respectively. With combined detection of tumor markers, the diagnostic power of those 4 indices was best, with an area under the ROC curve of 0.913 (95%CI 0.866~0.985), a sensitivity of 88.9% and a diagnostic accuracy of 90.4%. Conclusions: Combined detection of serum CEA, CA72-4, CA19-9 and TSGF increases the sensitivity and accuracy in diagnosis of GC, so it can be regarded as the important means for early diagnosis.

Immunotoxicity of Organophosphorous Pesticides, Pirimiphos-methyl and Methidathion in Balb/c Mice (Balb/c 마우스에서 유기인계 농약인 Pirimiphos-methyl 및 Methidathion의 면역독성)

  • Eam Juno H.;Chung Seung-Tae;Park Jae Hyun;Kil Jung Hyun;Lee Jong Kwon;Oh Hye Young;Kim Hyung Soo
    • Toxicological Research
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    • v.20 no.4
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    • pp.329-337
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    • 2004
  • Primiphos-methyl and methidathion as organophosphorus (OP) pesticides were tested for their immunotoxic effects on Balb/c mice. Three dose levels of primiphos-methyl (10, 60, or 120 mg/kg/day) and methidathion (0.5, 2.5 or 5.0 mg/kg/day) were administered orally in the mice for 4 weeks. After, changes in body weight gain, relative weight of spleen and thymus, viable splenic cell numbers, surface marker on immune cell, and proliferation activity were investigated. Results showed that neither Pirimiphos-methyl nor methidathion dosages changed significantly body weight, relative thymus and spleen weight, and thymus and spleen cellularities of the mice, but high dose treatment (120 mg/kg) of pirimiphos-methyl significantly decreased relative spleen weight and spleen cellularity of the mice. No alterations were observed in changes of LPS-proliferation response of splenocytes by exposure to any dose of pirimiphos-methyl and methidathion. However, pirimiphos-methyl dosages reduced ConA-proliferation response of splenocytes and both methidathion and pirimiphos-methyl decreased the ability of antibody production to SRBC. The results indicate that 28 days exposure to the high dose of pirimiphos-methyl suppress the function of splenic T and B cell function, and methidathion reduce the immune responsibility of B cell in mice without the changes in lymphoid organ weight or viability of splenocytes. Pirimiphos-methyl is more immunotoxic than methidathion although this has higher general toxicity than that.

Nuclear factor kappa-B- and activator protein-1-mediated immunostimulatory activity of compound K in monocytes and macrophages

  • Yang, Woo Seok;Yi, Young-Su;Kim, Donghyun;Kim, Min Ho;Park, Jae Gwang;Kim, Eunji;Lee, Sang Yeol;Yoon, Keejung;Kim, Jong-Hoon;Park, Junseong;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.298-306
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    • 2017
  • Background: Compound K (CK) is a bioactive derivative of ginsenoside Rb1 in Panax ginseng (Korean ginseng). Its biological and pharmacological activities have been studied in various disease conditions, although its immunomodulatory role in innate immunity mediated by monocytes/macrophages has been poorly understood. In this study, we aimed to elucidate the regulatory role of CK on cellular events mediated by monocytes and macrophages in innate immune responses. Methods: The immunomodulatory role of CK was explored by various immunoassays including cell-cell adhesion, fibronectin adhesion, cell migration, phagocytic uptake, costimulatory molecules, reactive oxygen species production, luciferase activity, and by the measurement of mRNA levels of proinflammatory genes. Results: Compound K induced cell cluster formation through cell-cell adhesion, cell migration, and phagocytic activity, but it suppressed cell-tissue interactions in U937 and RAW264.7 cells. Compound K also upregulated the surface expression of the cell adhesion molecule cluster of differentiation (CD) 43 (CD43) and costimulatory molecules CD69, CD80, and CD86, but it downregulated the expression of monocyte differentiation marker CD82 in RAW264.7 cells. Moreover, CK induced the release of reactive oxygen species and induced messenger RNA expression of proinflammatory genes, inducible nitric oxide synthase, and tumor necrosis factor-alpha by enhancing the nuclear translocation and transcriptional activities of nuclear factor kappa-B and activator protein-1. Conclusion: Our results suggest that CK has an immunomodulatory role in innate immune responses through regulating various cellular events mediated by monocytes and macrophages.

CRISPR/Cas9-mediated generation of a Plac8 knockout mouse model

  • Lee, HyunJeong;Kim, Joo-Il;Park, Jin-Sung;Roh, Jae-il;Lee, Jaehoon;Kang, Byeong-Cheol;Lee, Han-Woong
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.279-287
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    • 2018
  • Placenta specific 8 (PLAC8, also known as ONZIN) is a multi-functional protein that is highly expressed in the intestine, lung, spleen, and innate immune cells, and is involved in various diseases, including cancers, obesity, and innate immune deficiency. Here, we generated a Plac8 knockout mouse using the CRISPR/Cas9 system. The Cas9 mRNA and two single guide RNAs targeting a region near the translation start codon at Plac8 exon 2 were microinjected into mouse zygotes. This successfully eliminated the conventional translation start site, as confirmed by Sanger sequencing and PCR genotyping analysis. Unlike the previous Plac8 deficient models displaying increased adipose tissue and body weights, our male Plac8 knockout mice showed rather lower body weight than sex-matched littermate controls, though the only difference between these two mouse models is genetic context. Differently from the previously constructed embryonic stem cell-derived Plac8 knockout mouse that contains a neomycin resistance cassette, this knockout mouse model is free from a negative selection marker or other external insertions, which will be useful in future studies aimed at elucidating the multi-functional and physiological roles of PLAC8 in various diseases, without interference from exogenous foreign DNA.

Effects of Nature Herb Mixture Containing Laminaria japonica Areschoung on Fine Dust-induced Bronchitis in Mice (미세먼지로 유도된 기관지염증에 다시마가 함유된 복합물의 효과)

  • Han, Su-Hyun;Joo, In-Hwan;Lee, Su-Bin;Kim, Dong-Hee;Hyun, Ju-Myung;Park, Jong-Min
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.34 no.5
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    • pp.245-254
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    • 2020
  • The purpose of this study was to investigate the effects of natural herb mixture containing Laminaria japonica Areschoung (LAM) on fine dust-induced bronchitis in mice. Laminaria japonica Areschoung is main content of LAM, which is including fucoidan. Fucoidan extracted from phaophyta is known to prevent bronchitis and to defend against bacteria and virus infection. In this study, we experienced the effect of LAM on bronchitis and investigated gene expressions (e.g ; IL-8, COX-2, MCP-1) and bio-markers (e.g ; IL-8, PGE2, MUC5AC) associated with bronchitis by using A549 cells. Also, we investigated whether LAM can suppress the bronchitis in fine dust-induced animal models. We injected fine dust (50 ㎕) twice as INT (Intra-Nasal-Trachea) method. Then LAM (200 mg/kg and 400 mg/kg) were oral administered for 14 days. We analyzed the number of immune cells, immunoglobulin E, bio-markers level associated with bronchitis. LAM significantly decreased bio-marker (IL-1β, IL-6, IL-8, TNF-α, Histamine, PGE2), immune cells (white blood cell, neutrophil, lymphocyte, monocyte), and immunoglobulin E, that are increased by fine dust. Taken together, this study suggest that LAM can be used as effective herbal extract for bronchitis.

Dec2 inhibits macrophage pyroptosis to promote periodontal homeostasis

  • He, Dawei;Li, Xiaoyan;Zhang, Fengzhu;Wang, Chen;Liu, Yi;Bhawal, Ujjal K.;Sun, Jiang
    • Journal of Periodontal and Implant Science
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    • v.52 no.1
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    • pp.28-38
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    • 2022
  • Purpose: Macrophages play crucial roles as early responders to bacterial pathogens and promote/ or impede chronic inflammation in various tissues. Periodontal macrophage-induced pyroptosis results in physiological and pathological inflammatory responses. The transcription factor Dec2 is involved in regulating immune function and inflammatory processes. To characterize the potential unknown role of Dec2 in the innate immune system, we sought to elucidate the mechanism that may alleviate macrophage pyroptosis in periodontal inflammation. Methods: Porphyromonas gingivalis lipopolysaccharide (LPS) was used to induce pyroptosis in RAW 264.7 macrophages. Subsequently, we established an LPS-stimulated Dec2 overexpression cellular model in macrophages. Human chronic periodontitis tissues were employed to evaluate potential changes in inflammatory marker expression and pyroptosis. Finally, the effects of Dec2 deficiency on inflammation and pyroptosis were characterized in a P. gingivalis-treated experimental periodontitis Dec2-knockout mouse model. Results: Macrophages treated with LPS revealed significantly increased messenger RNA expression levels of Dec2 and interleukin (IL)-1β. Dec2 overexpression reduced IL-1β expression in macrophages treated with LPS. Overexpression of Dec2 also repressed the cleavage of gasdermin D (GSDMD), and the expression of caspase-11 was concurrently reduced in macrophages treated with LPS. Human chronic periodontitis tissues showed significantly higher gingival inflammation and pyroptosis-related protein expression than non-periodontitis tissues. In vivo, P. gingivalis-challenged mice exhibited a significant augmentation of F4/80, tumor necrosis factor-α, and IL-1β. Dec2 deficiency markedly induced GSDMD expression in the periodontal ligament of P. gingivalis-challenged mice. Conclusions: Our findings indicate that Dec2 deficiency exacerbated P. gingivalis LPS-induced periodontal inflammation and GSDMD-mediated pyroptosis. Collectively, our results present novel insights into the molecular functions of macrophage pyroptosis and document an unforeseen role of Dec2 in pyroptosis.

Development of Tumor Screening ELISA kit by Using Novel Tumor Antigen, CM1 (새로운 종양항원인 CM1을 이용한 종양 진단 ELISA kit의 개발)

  • Kang, Jae-Seung;Kim, Dae-Jin;Kim, Young-In;Chang, Ka-Young;Lee, Wang-Jae
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.124-129
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    • 2005
  • Background: CM1 (centrocyte/-blast marker 1) is originally defined as a germinal center B cell marker. It is known that CM1 plays a critical role on B cell development in germinal center. In addition, we have found that CM1 is expressed on lymphoma cell lines, such as Raji, Ramos and IM-9. This means that CM1 might be served as a tumor marker as well. In the present study, we examined the expression of CM1 on the surface of the other tumors and the possibility of the development of tumor screening ELISA kit by using CM1. Methods: First, we have examined the expression of CM1 on stomach cancer and hepatoma, which are predominantly (discovered) occurred in Korean, by flow cytometry analysis. After purifying of CM1 antigen from Raji and Ramos, the optimal ELISA condition was determined. And then we compared the level of CM1 between normal individuals and cancer patients by ELISA. To decrease the non-specific binding of anti-CM1 mAb with serum components except CM1 and to enhance the diagnostic accuracy, albumin depletion spin column was used. Results: CM1 was highly expressed on stomach cancer and hepatoma cell lines. In addition, we have also confirmed the increased CM1 expression on cancer patients. The difference of CM1 expression between normal individuals and cancer patients were more clearly observed, after deletion of serum albumin by using albumin depletion spin column. Conclusion: Based on the results from this study, CM1 might be a useful molecule for the early diagnosis of cancer. In addition, further studies for the increase of ELISA sensitivity and appropriate albumin depletion methods should be needed.