• Title/Summary/Keyword: Hemagglutination activity

Search Result 67, Processing Time 0.031 seconds

Production of Monoclonal Antibody to Newcastle Disease Virus and its Diagnostic Use (뉴켓슬병 바이러스에 대한 단크론성 항체생산 및 진단에 이용)

  • Chung Ok Choi;Chung Gil Lee;Sung Man Cho;Jin Soo Na;Soo Hwan An;Joon Hun Kwon
    • Korean Journal of Poultry Science
    • /
    • v.15 no.3
    • /
    • pp.207-210
    • /
    • 1988
  • A total of 3 hybridoma clones producting menoclonal antibody (MCA) against Newcastle disease virus(NDV) was raised by cell fusion method. The MCAs did not cross react against other avian or mammalian viruses tested. However, these antibodies reacted with all strains of velogenic and lentogenic NDVs tested indicating that they are unable to discriminate the possible antigenic differences among NDVs. All. the MCAs were classified as IgG type and did not show neutralizing and hemagglutination inhibition (HAI) activity except one clone which has low HAI activity. One of these MCA raised in mouse ascites revealed the titer of $10^6$ by indirect immunofluorescent antibody (IFA) test Using the MCA, virulent NDV could easily be detected from tracheal and conjunctival smears made 2 to 3 days after experimental infection.

  • PDF

Insect Cell Surface Expression of Hemagglutinin (HA) of Egyptian H5N1 Avian Influenza Virus Under Transcriptional Control of Whispovirus Immediate Early-1 Promoter

  • Gadalla, M.R.;El-Deeb, A.H.;Emara, M.M.;Hussein, H.A.
    • Journal of Microbiology and Biotechnology
    • /
    • v.24 no.12
    • /
    • pp.1719-1727
    • /
    • 2014
  • In the present study, whispovirus immediate early 1 promoter (ie-1) was used to initiate surface expression of the hemagglutinin (HA) protein of Egyptian H5N1 avian influenza virus (AIV) by using the baculovirus expression vector system. The HA gene and whispovirus ie-1 promoter sequence were synthesized as a fused expression cassette (ie1-HA) and successfully cloned into the pFastBac-1 transfer vector. The recombinant vector was transformed into DH10Bac competent cells, and the recombinant bacmid was generated via site-specific transposition. The recombinant bacmid was used for transfection of Spodoptera frugiperda (Sf-9) insect cells to construct the recombinant baculovirus and to induce expression of the HA protein of H5N1 AIV. The recombinant glycoprotein expressed in Sf-9 cells showed hemadsorption activity. Hemagglutination activity was also detected in both extra- and intracellular recombinant HAs. Both the HA and hemadsorption activities were inhibited by reference polyclonal anti-H5 sera. Significant expression of the recombinant protein was observed on the surface of infected insect cells by using immunofluorescence. SDS-PAGE analysis of the expressed protein revealed the presence of a visually distinguishable band of ~63 kDa in size, which was absent in the non-infected cell control. Western blot analysis confirmed that the distinct 63 kDa band corresponded to the recombinant HA glycoprotein of H5N1 AIV. This study reports the successful expression of the HA protein of H5N1 AIV. The expressed protein was displayed on the plasma membrane of infected insect cells under the control of whispovirus ie-1 promoter by using the baculovirus expression vector system.

Effect of Terminalia chebula on Physiological Activity in Mice (가자(Terminalia chebula) 추출물이 마우스의 생리활성에 미치는 영향)

  • 박종옥;이승은
    • Journal of Life Science
    • /
    • v.14 no.1
    • /
    • pp.148-153
    • /
    • 2004
  • In this study, we investigated the effect of water, extract of Terminalia Chebula (TC) on physiological activity in mice. TC water extract showed hemagglutination against several different types of red blood cells. $LD_{50}$ of TC extract was 390 mg/kg (po). Treatment of TC water extract orally administered 200, 300 mg/kg daily for one week. Hepatic cytosolic enzymes, xanthine oxidase and aldehyde oxidase activities were significantly increased comparison with normal group. Treatment of TC water extract increased hepatic malondialdehyde (MDA) formation, and reduced glutathione content. We also found that the decreased activities of glutathione S-transferase and glutathione reductase but was not affected activities of $\gamma$-glutamylcysteine synthetase after treatment of TC water extract. These results suggested that increase of the hepatic lipid peroxide is caused by glutathione reduction.

Immunobiological Studies on Doses of Methanol Extract of Astragali Radix (황기의 메탄올 추출물의 용량에 따른 면역생물학적 연구)

  • Kim, Joung-Hoon;Park, Joung-Suk;Chae, Byeong-Suk;Kang, Tae-Wook;Park, Chan-Bong;Ahn, Young-Keun
    • YAKHAK HOEJI
    • /
    • v.40 no.3
    • /
    • pp.326-334
    • /
    • 1996
  • Effects of methanol extract of Astragali Radix (AR) on the immune responses were studied using ICR mice. Mice were divided into 4 groups (10mice/group), and methanol extracts of AR at doses of 0.05, 0.25 and 1.25g/kg were orally administered to ICR mice once a day for 2 weeks. Mice were immunized and challenged with sheep red blood cells (SRBC). The results of this study were summarized as follows; (1) Methanol extract of AR at 0.05, 0.25 and 1.25g/kg didn't affect the weight ratios of thymus to body, as compared with those in controls, but significantly increased spleen weight ratio. (2)Methanol extract of AR at 0.05 and 0.25g/kg significantly increased hemagglutination titer and splenic plaque forming cells corresponding to humoral immunity, as compared with those in controls, but their enhancements were somewhat lowered at a high dose (1.25g/kg). (3) Methanol extract of AR at 0.05 and 0.25g/kg siginificantly increased delayed-type hypersensitivity reaction resulted from cell-mediated immunity, as compared with those in controls, but not so significant increases were observed at a high dose (1.25g/kg). (4) Methanol extract of AR at 0.05 and 0.25g/kg significantly increased phagocytic activity and the number of circulating leukocyte compared with those in controls, but their enhancements were lowered at a high dose (1.25g/kg). These results suggest that methanol extract of Astragali Radix increased humoral and cell-mediated immune responses, phagocytic activity and the number of circulating leukocyte, dependent upon dose, but inhibited their enhancement effects were decreased at a high dose (1.25g/kg).

  • PDF

Biochemical Properties of Seed Lectin from Korean Soybean Cultivars Developed for Soy Source (한국산 장류콩 종자 렉틴의 생화학적 특성)

  • Wang, Yushan;Roh, Kwang-Soo
    • KSBB Journal
    • /
    • v.24 no.2
    • /
    • pp.170-176
    • /
    • 2009
  • Lectin was finally isolated on Sephadex G-100 from Korean soybean cultivars developed for soy source and investigated its some biochemical properties. Native PAGE pattern of this lectin revealed a molecular weight of 108 kDa as tetramer. The molecular weight of this lectin isolated as double protein band by SDS-PAGE was calculated to be 32 and 22 kDa from the relative mobilities compared with those of the standard proteins. Among the tested red blood cell, the isolated lectin agglutinated rabbit red blood cell treated with trypsin, but did not agglutinated human red blood cells (A, B, AB, O), rat, and untreated rabbit red blood cell. The optimal temperature and thermal stability of isolated lectin was at 20-$50^{\circ}C$ and 10-$60^{\circ}C$, respectively. This lectin was stable at 7.2, and showed complete loss in its activity below pH 6.2 and above pH 8.0.

Effects of Methionine Diets on the Immunotoxicity of Ethanol in ICR Mice (생쥐에 있어서 에탄올의 면역독성에 미치는 메치오닌 식이의 영향)

  • Ahn, Young-Keun;Kim, Joung-Hoon;Koo, Gi-Bum;Moon, Jae-Gyu
    • YAKHAK HOEJI
    • /
    • v.36 no.4
    • /
    • pp.303-314
    • /
    • 1992
  • Experiments were performed on mice to investigate the effect of methionine diets (MET) on the immunotoxicity of ethanol. ICR female mice were divided into 5 groups, Met (Basal (B)+0.19% methionine(M), B+1.71% M and B+5.13%W) and ethanol(4%) were administered ad libitum for 21 days. The mice were evaluated for changes in immune status as measured by antibody titer, Arthus reaction, delayed type hypersensitivity (DTH), rosette forming cell(RFC) and plaque forming cell (PFC) to sheep red blood cells (S-RBC). To investigate the change of the non-specific immune response, the number of leukocytes in peripheral blood and phagocyte activity were measured. The results were summarized as follows: (1) The weight ratios of spleen and thymus to body weight were significantly increased by the B+0.19% M, B+0.57% M and B+1.71% M groups in comparison with control group(B), but B+5.13% M group was significantly decreased. (2) Humoral immune responses were significantly increased by the B+0.19% M and B+0.57% M groups in comparison with control group, but B+5.13% M group was significantly decreased. (3) Cellular immune responses were significantly decreased by the B+1.71% M and B+5.13% M groups in comparison with control group. (4) Phagocyte activities were significantly increased by the B+0.19% M, B+0.57% M and B+1.71% M groups in comparison with control groups, but B+5.13% M group was significantly decreased. (5) The number of circulating leukocyte was significantly increased in the B+0.19% M and B+0.57% M groups in comparison with control group, but B+5.13% M group was significantly decreased.

  • PDF

Binding Specificity of Philyra pisum Lectin to Pathogen-Associated Molecular Patterns, and Its Secondary Structure

  • Park, Byung Tae;Kim, Byung Sun;Park, Heajin;Jeong, Jaehoon;Hyun, Hanbit;Hwang, Hye Seong;Kim, Ha Hyung
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.17 no.6
    • /
    • pp.547-551
    • /
    • 2013
  • We recently reported a Philyra pisum lectin (PPL) that exerts mitogenic effects on human lymphocytes, and its molecular characterization. The present study provides a more detailed characterization of PPL based on the results from a monosaccharide analysis indicating that PPL is a glycoprotein, and circular dichroism spectra revealing its estimated ${\alpha}$-helix, ${\beta}$-sheet, ${\beta}$-turn, and random coil contents to be 14.0%, 39.6%, 15.8%, and 30.6%, respectively. These contents are quite similar to those of deglycosylated PPL, indicating that glycans do not affect its intact structure. The binding properties to different pathogen-associated molecular patterns were investigated with hemagglutination inhibition assays using lipoteichoic acid from Gram-positive bacteria, lipopolysaccharide from Gram-negative bacteria, and both mannan and ${\beta}$-1,3-glucan from fungi. PPL binds to lipoteichoic acids and mannan, but not to lipopolysaccharides or ${\beta}$-1,3-glucan. PPL exerted no significant antiproliferative effects against human breast or bladder cancer cells. These results indicate that PPL is a glycoprotein with a lipoteichoic acid or mannan-binding specificity and which contains low and high proportions of ${\alpha}$-helix and ${\beta}$-structures, respectively. These properties are inherent to the innate immune system of P. pisum and indicate that PPL could be involved in signal transmission into Gram-positive bacteria or fungi.

The Effects of Several Types of Bibimbabs on Immune Activities in Mice (여러 가지 비빔밥의 섭취가 생쥐의 각종 면역 활성에 미치는 효과)

  • Kim, Nam-Seok;Cho, Mun-Ku;Oh, Suk-Heung;Choi, Dong-Seong;Jung, Mun-Yhung;Woo, Ja-Won;Kwon, Jin;Kim, Dong-Hun;Oh, Chan-Ho
    • Journal of the East Asian Society of Dietary Life
    • /
    • v.23 no.1
    • /
    • pp.23-30
    • /
    • 2013
  • The purpose of this study was to evaluate the effects of several types of bibimbab (a Korean traditional meal of mixed rice with assorted vegetables), on various immune activities. Compared to control animals in a mouse model (given hamburgers), the oral administration of a portion of bibimbab containing wild plants significantly increased splenic B/T, thymic Th lymphocyte subpopulations, serum IFN-${\gamma}$ production, and enhanced hemagglutination titers up to 300%. Also, a consumption of mushroom-bulgogi bibimbab and Jeonju-style bibimbab markedly decreased compound 48/80-induced systemic anaphylaxis (immediate hypersensitivity), while bibimbab with wild plants inhibited SRBC-induced delayed type hypersensitivity. These results suggest that bibimbab with wild plants both up-regulate on immune activities and have anti-allergenic properties.

Novel Phage Display-Derived H5N1-Specific scFvs with Potential Use in Rapid Avian Flu Diagnosis

  • Wu, Jie;Zeng, Xian-Qiao;Zhang, Hong-Bin;Ni, Han-Zhong;Pei, Lei;Zou, Li-Rong;Liang, Li-Jun;Zhang, Xin;Lin, Jin-Yan;Ke, Chang-Wen
    • Journal of Microbiology and Biotechnology
    • /
    • v.24 no.5
    • /
    • pp.704-713
    • /
    • 2014
  • The highly pathogenic avian influenza A (HPAI) viruses of the H5N1 subtype infect poultry and have also been spreading to humans. Although new antiviral drugs and vaccinations can be effective, rapid detection would be more efficient to control the outbreak of infections. In this study, a phage-display library was applied to select antibody fragments for HPAI strain A/Hubei/1/2010. As a result, three clones were selected and sequenced. A hemagglutinin inhibition assay of the three scFvs revealed that none exhibited hemagglutination inhibition activity towards the H5N1 virus, yet they showed a higher binding affinity for several HPAI H5N1 strains compared with other influenza viruses. An ELISA confirmed that the HA protein was the target of the scFvs, and the results of a protein structure simulation showed that all the selected scFvs bound to the HA2 subunit of the HA protein. In conclusion, the three selected scFVs could be useful for developing a specific detection tool for the surveillance of HPAI epidemic strains.

Isolation and identification of mammalian orthoreovirus type 3 from a Korean roe deer (Capreolus pygargus)

  • Yang, Dong-Kun;An, Sungjun;Park, Yeseul;Yoo, Jae Young;Park, Yu-Ri;Park, Jungwon;Kim, Jong-Taek;Ahn, Sangjin;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
    • /
    • v.61 no.2
    • /
    • pp.13.1-13.8
    • /
    • 2021
  • Mammalian reovirus (MRV) causes respiratory and intestinal disease in mammals. Although MRV isolates have been reported to circulate in several animals, there are no reports on Korean MRV isolates from wildlife. We investigated the biological and molecular characteristics of Korean MRV isolates based on the nucleotide sequence of the segment 1 gene. In total, 144 swabs from wild animals were prepared for virus isolation. Based on virus isolation with specific cytopathic effects, indirect fluorescence assays, electron microscopy, and reverse transcription-polymerase chain reaction, only one isolate was confirmed to be MRV from a Korean roe deer (Capreolus pygargus). The isolate exhibited a hemagglutination activity level of 16 units with pig erythrocytes and had a maximum viral titer of 105.7 50% tissue culture infectious dose (TCID50)/mL in Vero cells at 5 days after inoculation. The nucleotide and amino-acid sequences of the partial segment S1 of the MReo2045 isolate were determined and compared with those of other MRV strains. The MReo2045 isolate had nucleotide sequences similar to MRV-3 and was most similar (96.1%) to the T3/Bat/Germany/342/08 strain, which was isolated in Germany in 2008. The MReo2045 isolate will be useful as an antigen for sero-epidemiological studies and developing diagnostic tools.