• Title/Summary/Keyword: ELISA antibody

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Detection of Helicobacter pylori Infection Rate in Resected Gastric Cancer Patients (위절제술을 받은 위암 환자의 Helicobacter pylori 감염률 측정에 관한 연구)

  • Kim Sung;Noh Jae-Hyung;Kim Sung;Kim Yong-Il;Kim Jin-Pok
    • Journal of Gastric Cancer
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    • v.3 no.2
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    • pp.97-103
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    • 2003
  • Purpose: This study aims to determine the infection rate of Helicobacter pylori (H. pylori) in gastric cancer patients who received gastrectomies, and to compare the rates of H.pylori infection detected by serological test and that of histopathological test, and to evaluate its clinical meaning. Materials and Methods: Fifty two patients were selected from those who underwent gastrectomies at the Department of Surgery, Samsung Medical Center, from March 1997 to May 1997. The control group consisted of healthy 103 persons visited the center for health promotion in Samsung Medical Center. In both groups, we quantitatively checked serum level of IgG anti H. pylori antibody titer by ELISA using GAP IgG test kit (BioRad, USA) for the serological test, and we microscopically examined the surgical specimen stained by Warthin-Starry silver staining method for the histopathological test. Results: The seropositive rate of H. pylori in the patients' group was $71.2\%$ (37/52), and the control group was $65.0\%$ (67/103). The difference between two groups was statistically significant. However the histopathological study showed that the overall detection rate of H. pylori was $61.5\%$ (32/52) in the patients' group and $61.2\%$ (63/103) in the control group; nd this difference was not statistically significant Conclusion: We could confirm that H.pylori infection rate in the gastric cancer resected patients was statistically higher than in the normal healthy persons even in small population. And the detection method for the H. pylori infection by serological test was presumed to be better than that of histopathological test using surgical specimen. Further study for the larger population by well-organized multicenters will be needed.

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Biological Functions of Lactoferrin and Antigenic Determinants of Lactoferricin (Lactoferrin의 생리적 기능과 Lactoferricin의 항원결정기)

  • Nam, Myeong-Su;Shimazaki, K.
    • Journal of Dairy Science and Biotechnology
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    • v.14 no.2
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    • pp.229-245
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    • 1996
  • 1. CM-Sephadex C-50-120 column을 사용하여 HLF를 효과적으로 정제하였다. CM-Sepha-dex 50-120 column chromatography를 수행한 결과, HLF는 NaCl 350 mM-500 mM 사이에서 용출되었으며, 용출된 분획을 SDS-PAGE를 수행해서 단일 band를 확인하였다. Anti-HLF antibody를 이용한 Western blotting 결과 nitrocellulose paper 상에 단일 band 가 나타나므로 HLF 가 효과적으로 정제되었다는 것을 알 수 있었다. 2. Con A, PWM, PWA LPS 등의 자극원으로 단핵구와 대식세포를 자극한 다음 BLF를처리한 배 양액을 IL-1 bioassy한 결과는 Con A 33%, PMA 33%, PWM 15%, LPS 35% 이고, HLF로 처리하여 IL-1 bioassay를 한 결과는 Con A 15%, PMA 22%, PWM 10%, LPS 5%로 감소된 것으로 나타났다. 3. K-562 세포를 이용한 colony forming assay에서 BLF가 10 g/ml일 때는 30%, 30 g/ml일 때는 35%, HLF는 10 g/ml일 때는 5%,30 g/ml일 때는 30%의 저해를 나타냈다. 4. Lactoferrin의 면역증강효과를 알아보기 위하여 hapten인 VCR-BSA를 투여 한 후, 생성되는 항체생성능력을 ELISA로 비교하였다. 그 결과 HLF 및 BLF 투여군에서 대조군에 비하여 adjuvant 효과를 관찰할 수 있었다. 또한 이때 macrophage 수는 대조군에 비하여 HLF와 BLF를 투여한 군이 증가됨을 알 수 있었다. 이러한 결과로 미루어 볼 때 LF는 macrophage를 활성화 시켜서 항체 생성 능력을 증가시키는 효과가 있음을 관찰할 수 있었다. 5. Balb/c mouse의 thymus로부터 분리한 CD4- CD8- 세포를 BLF로 처리하여 24 시간 배양한 후 CB4$^-$ CD8$^-$ 세포의 분화를 측정한 결과, CD4$^-$를 CD4$^+$ 로 분화하였다. 그리고 HLF로 처리하여 24 시간 배양 후 CD4$^-$ CD8$^-$ 세포의 분화를 측정한 결과, CD4$^-$ CD8$^-$를 CD4$^+$ CD8$^+$ 로 분화하였다. 6. Lactoferrin이 T cell의 IL-2 production에 미치는 영향은 PMA 처리군, PMA+OKT3처리군, LF 단독 처리군 보다 PMA+OKT3+LF를 처리한 군이 IL-2 생성에 영향을 미쳤다. 7. Lfcin B의 단일크론항체에 의해 인식되어지는 Lfcin B의 항원결정기는 ‘QWR’로 밝혀졌다.

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Single step purification of potent antigenic protein from sparganum by gelatin-affinity chromatography (젤라틴 친화성 크로마토그래피를 이용한 스파르가눔 성분단백질의 순수분리)

  • Yoon Kong;Shin-Yong Kang;Seung-Yull Cho
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.1-8
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    • 1991
  • Out of many component proteins in crude saline extract of Spirometra mansoni plerocercoid (sparganum) , 36 kDa and 29 kDa proteins were found to be the most antigenic and were already purified by immunoaffinity chromatography using monoclonal antibody as a ligand. In this study, a single step purification of these potent antigenic proteins of sparganum extract was investigated. When the crude saline extract was charged to gelatin-Sepharose 4B affinity column, 36 kDa and 29 kDa protein fractions were bound. SDS-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE/immunoblot confirmed that the bound protein to gelatin was serologically pure. When evaluated by ELISA with patients sera, the purified protein of 36 and 29 kDa also showed improved antigenicity.

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Seroprevalence of Pertussis in Healthcare Workers without Adult Pertussis Vaccine Use at a University Hospital in Korea

  • Choi, Won Suk;Kim, Su Hyun;Park, Dae Won
    • Journal of Korean Medical Science
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    • v.33 no.50
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    • pp.321.1-321.7
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    • 2018
  • Background: Pertussis is highly contagious respiratory disease. Healthcare workers (HCWs) can be an important mediator of the disease. A seroprevalence of pertussis was investigated in HCWs to determine the immune status against pertussis and to detect the unidentified pertussis. Methods: This study was conducted for HCWs at a hospital located in Korea in 2011. After obtaining written informed consent for HCWs voluntarily participating in the study, 10 mL of blood was collected from each subject. Demographic and medical data were collected using questionnaire. Data on the underlying disease and vaccination history were reviewed again through medical records. The presence of anti-pertussis toxin (anti-PT) immunoglobulin G (IgG), and immunoglobulin A (IgA) was detected by quantitative analysis using a commercially available ELISA kit (EUROIMMUN, $L{\ddot{u}}beck$, Germany). Results: A total of 412 HCW participated in the study. Among them, 14 were excluded due to the inadequate sample amount or medical history not secured. Of the 398 HCWs analyzed, 16.6% (66/398) were men and the mean age was $33.82{\pm}9.10years$ (range, 21-67). The mean anti-PT IgG titer was $8.32{\pm}20.40IU/mL$ (range, 0.4-287.5 IU/mL). The overall seroprevalence (rate of anti-PT IgG antibody [Ab] titer > 5 IU/mL) was 33.7%. Three (0.8%) HCWs had the Ab level > 100 IU/mL indicated acute infection. There was no statistically significant difference in the seroprevalence and mean titer of anti-PT IgG Ab according to age group, type of occupation, patient-facing position, or working in the pediatric department. Conclusion: The seroprevalence of pertussis of the HCWs of a university hospital in Korea was low, and there were some unrecognized acute infections. Therefore, booster immunization of pertussis to HCWs should be actively considered.

Inhibitory Effect of Chitosan and Phosphate Cross-linked Chitosan against Cucumber Mosaic Virus and Pepper Mild Mottle Virus

  • Gangireddygari, Venkata Subba Reddy;Chung, Bong Nam;Cho, In-Sook;Yoon, Ju-Yeon
    • The Plant Pathology Journal
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    • v.37 no.6
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    • pp.632-640
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    • 2021
  • Cucumber mosaic virus (CMV) and Pepper mild mottle virus (PMMoV) causes severe economic loss in crop productivity of both agriculture and horticulture crops in Korea. The previous surveys showed that naturally available biopolymer material - chitosan (CS), which is from shrimp cells, reduced CMV accumulation on pepper. To improve the antiviral activity of CS, it was synthesized to form phosphate cross-linked chitosan (PCS) and compared with the original CS. Initially, the activity of CS and PCS (0.01%, 0.05%, and 0.1% concentration) compound against PMMoV infection and replication was tested using a half-leaf assay on Nicotiana glutinosa leaves. The total number of local lesions represented on a leaf of N. glutinosa were counted and analyzed with phosphate buffer treated leaves as a negative control. The leaves treated with a 0.1% concentration of CS or PCS compounds exhibited an inhibition effect by 40-75% compared with the control leaves. The same treatment significantly reduced about 40% CMV accumulation measured by double antibody sandwich enzyme-linked immunosorbent assay and increased the relative expression levels of the NPR1, PR-1, cysteine protease inhibitor gene, LOX, PAL, SRC2, CRF3 and ERF4 genes analyzed by quantitative reverse transcriptase-polymerase chain reaction, in chili pepper plants.

Inhibitory Effects of Pepper Mild Mottle Virus Infection by Supernatants of Five Bacterial Cultures in Capsicum annuum L.

  • Venkata Subba Reddy, Gangireddygari;In-Sook, Cho;Sena, Choi;Ju-Yeon, Yoon
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.646-655
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    • 2022
  • Pepper mild mottle virus (PMMoV), one of the most prevalent viruses in chili pepper (Capsicum annuum L.) is a non-enveloped, rod-shaped, single-stranded positive-sense RNA virus classified in the genus Tobamovirus. The supernatants of five bacterial cultures (Pseudomonas putida [PP], Bacillus licheniformis [BLI], P. fluorescens [PF], Serratia marcescens [SER], and B. amyloliquifaciens [BA]) were analyzed to find novel antiviral agents to PMMoV in chili pepper. Foliar spraying with supernatants (1:1, v/v) obtained from Luria-Bertani broth cultures of PP, BLI, PF, SER, and BA inhibited PMMoV infection of chili pepper if applied before the PMMoV inoculation. Double-antibody sandwich enzyme-linked immunosorbent assay showed that treatments of five supernatants resulted in 51-66% reductions in PMMoV accumulation in the treated chili pepper. To identify key compounds in supernatants of PP, BLI, PF, SER, and BA, the supernatants were subjected to gas chromatography-mass spectrometry. The 24 different types of compounds were identified from the supernatants of PP, BLI, PF, SER, and BA. The compounds vary from supernatants of one bacterial culture to another which includes simple compounds-alkanes, ketones, alcohols, and an aromatic ring containing compounds. The compounds triggered the inhibitory effect on PMMoV propagation in chili pepper plants. In conclusion, the cultures could be used to further conduct tissue culture and field trial experiments as potential bio-control agents.

Inhibitory Effect of a decoction composed of Evodia rutaecarpa (Juss.) Benth. and Chaenomeles sinensis Koehne and its component herbal medicines on Collagen II-induced Arthritis Mice (Collagen II-induced Arthritis 생쥐에 대한 오수유(吳茱萸), 목과(木瓜) 및 배합약물의 관절염 억제 효과)

  • Park, Dae-Jung;Lee, Young-Cheol;Lee, Jang-Cheon
    • The Korea Journal of Herbology
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    • v.29 no.4
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    • pp.35-44
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    • 2014
  • Objectives : Evodia rutaecarpa (Juss.) Benth.(ER) and Chaenomeles sinensis Koehne (CS) have multiple applications and were known to have anti-inflammatory effects. In the current study, we investigated to clearly understand the mechanism of therapeutic role for CS, ER and their combination in CIA model mice. Methods : DBA/1OlaHsd mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with CS, ER and their combination once a day for 7 weeks. Cytokine production and gene expression were assessed during CIA (collagen-induced arthritis) model mice in knee joint, lymph node (LN) using ELISA and FACS analysis. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. Result : Oral administration of CS, ER and their combination (150 mg/kg) significantly suppressed the progression of CIA, and significantly suppressed the progression of CIA and inhibited the production of TNF-${\alpha}$ and IL-6 in serum. The erosion of cartilage was dramatically reduced in mouse knees after treatment with CS plus ER. Conclusion : These result suggest that CS plus ER significantly suppressed the progression of CIA and that this action was characterized by the decreased production of TNF-${\alpha}$, IL-6 and collagen II specific antibody in serum.

Effects of Piperis Longi Fructus on Regulatory T Cells Number, IgE, Histamine Production in Asthma Model Mice and Th1/Th2 Cytokine Balance in vitro (천식 모델 생쥐에서 필발이 CD25+T 세포수, IgE, Histamine 생성량과 in vitro에서 Th1/Th2 Cytokine Balance에 미치는 영향)

  • Lee, Young-Cheol;Kim, Seung-Hyung
    • The Korea Journal of Herbology
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    • v.24 no.1
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    • pp.79-88
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    • 2009
  • Objectives : It has been recently shown that Piperis Longi Fructus (PLF) is involved in the reduction of eosinophil recruitment and production of Th2 cytokines in vivo. However, the main therapeutic mechanisms of PLF remains a matter of considerable debate. To investigate the therapeutic mechanisms of PLF, we examined the influence of PLF on regulatory T cells number, IgE, histamine production in vivo and Th1/Th2 cytokine balance in vitro. Methods : All mice were immunized on two different days (21 days and 7 days before inhalational exposure) by i.p. injections of 0.2 $m\ell$ alum-precipitated Ag containing 100 ${\mu}g$ of OVA bound to 4 mg of aluminum hydroxide in PBS. Seven days after the second sensitization, mice were exposed to aerosolized ovalbumin for 30 min/day on 3 days/week for 12 weeks(at a flow rate of 250 L/min, 2.5% ovalbumin in normal saline) and PLF (150 mg/kg) were orally administered 3 times a week for 8 weeks. Splenocytes from C57BL/6 mice at 8 weeks of age were stimulated with anti-CD3 (1 mg/ml) plus anti-CD28 (1 mg/ml) antibody for 48hrs. IL-4 and IFN-$\gamma$ in the culture supernatants were measured by ELISA Results : The suppressive effects of PLF on asthma model were demonstrated by the increase the number of regulatory T cells and by reducing IgE, histamine production in vivo and modulation of Th1/Th2 cytokine balance. Conclusions : These results indicate that PLF has a deep inhibitory effects on asthma model mice by increase the number of regulatory T cells, and by reducing IgE, histamine production.

Rehmannioside D mitigates disease progression in rats with experimental-induced diminished ovarian reserve via Forkhead Box O1/KLOTHO axis

  • Yan Liang;Huimin Wang;Jin Chen;Lingyan Chen;Xiaoyong Chen
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.2
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    • pp.167-176
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    • 2023
  • This study aims to explore the impact of Rehmannioside D (RD) on ovarian functions of rats with diminished ovarian reserve (DOR) and its underlying mechanisms of action. A single injection of cyclophosphamide was performed to establish a DOR rat model, and fourteen days after the injection, the rats were intragastrically administrated with RD for two weeks. Rat estrus cycles were tested using vaginal smears. Ovarian tissues were histologically evaluated, the number of primordial, mature, and atretic follicles was calculated, and the apoptotic rate of granulosa cells. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) levels were determined by ELISA assays. Protein levels of Forkhead Box O1 (FOXO1), KLOTHO, Bcl-2, and Bax were investigated in ovarian tissues of DOR rats. The binding between FOXO1 and KLOTHO was verified by ChIP assay. High-dose administration of RD into DOR rats improved their estrus cycles, increased ovarian index, enhanced the number of primordial and mature follicles, reduced the number of atretic follicle number, and ovarian granulosa cell apoptosis in addition to inhibiting FSH and LH levels and upregulating E2 expression. FOXO1 and KLOTHO were significantly suppressed in DOR rats. FOXO1 knockdown partially suppressed the protective effects of RD on DOR rats, and KLOTHO overexpression could restore RD-induced blockade of DOR development despite knocking down FOXO1. FOXO1 antibody enriched KLOTHO promoter, and the binding between them was reduced in DOR group compared to that in sham group. RD improved ovarian functions in DOR rats and diminished granulosa cell apoptosis via the FOXO1/KLOTHO axis.

Fucoidan Increases Porcine Neutrophil Extracellular Trap Formation through TNF-α from Peripheral Blood Mononuclear Cells

  • Changwoo Nahm;Yoonhoi Koo;Taesik Yun;Hakhyun Kim;Byeong-Teck Kang;Mhan-Pyo Yang
    • Journal of Veterinary Clinics
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    • v.40 no.3
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    • pp.175-181
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    • 2023
  • Fucoidan extracted from brown seaweed has a variety of biological activities. Neutrophil extracellular traps (NETs) formation is an immune response for the invasion of pathogens. Neutrophils release granule protein and chromatin that form extracellular fibers that bind microbes. These NETs degrade virulence factors and kill bacteria. The aim of this study was to investigate the effect of fucoidan on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation was determined by fluorescence emission of propidium iodide (PI) in PMNs by a fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α from peripheral blood mononuclear cells (PBMCs) was measured by ELISA method. Fucoidan itself did not show any direct effect on NET formation. However, NET formation of PMNs was increased by the culture supernatant from PBMCs treated with fucoidan. The NET formation of PMNs were also enhanced by treatment with recombinant porcine (rp) TNF-α. The ability of culture supernatant from PBMCs treated with fucoidan to increase the NET formation of PMNs was inhibited by addition of goat anti-rp TNF-α polyclonal antibody (pAb) (IgG) prior to the culture. The increase of NET formation by rp TNF-α was also inhibited by goat anti-rp TNF-α pAb (IgG). The level of TNF-α in culture supernatant from PBMCs was increased by treatment with fucoidan. These results suggest that fucoidan increases porcine NET formation, which is mediated by TNF-α produced from PBMCs.