• Title/Summary/Keyword: immunofluorescent antibodies

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The serological studies on infectious bursal disease (전염성 F낭병에 대한 혈청학적 연구)

  • 정영미;서석열;도홍기;조정곤;노수일
    • Korean Journal of Veterinary Service
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    • v.23 no.3
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    • pp.271-279
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    • 2000
  • This study was carried out to provide the fundamental information for development of proper vaccination program against infectious bursal disease(IBD) to the local chicken farms. The antigen detection was peformed from 8 samples of bursa of Fabricius with agar gel precipitation(AGP) and indirect immunofluorescent assay(IFA), And also, the antibodies in serum samples were detected by the various serological methods such as commercial ELISA assay, AGP and virus neutralization(VN) test. 1. The antigen detection rates were 25% for AGP which is 2 out of 8 farms and 10 out of 40 bursas, and 25% which Is 2 out of 8 farms and 20% 8 out of 40 bursas for IFA, respectively. 2. The mean titer of maternal antibody (>3,000) existed until 10 days of the age with ELISA-GMT. 3. The antibody positive rates which are over 80% showed until 5 days of the age with ELISA and at 10 days of the age with AGP except one, but none of them showed from 1 day of the age. This report came to conclusions that both the protective maternal antibody titers and the antigen positive rates were significant until at the 10 days of the age.

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Immunohistoehemical Observation on the Antigens Inducing IgG and IgM Antibodies against Sparganum (IgG와 IgM 항체를 유도하는 sparganum의 항원에 관한 면역조직화학적 및 전기영동에 의한 연구)

  • 김창환;최완성
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.339-354
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    • 1991
  • Localization and characterization of the antigenic components of sparganum which induced IgG and IsM antibodies in the host were studied by immunohistochemical techniques and SDS-PAGT and Western blotting. The antigen recognized by IgG antibody of rats or mice which were immunised by infection or injection of crude extracts of metacestodes of Spirometra erinacei, was located in the parenchyme of sparganum, especially at the cortex and around the calcareous corpuscles. The immunoreaction was demonstrated not only in the encysted fibrous wall of host but around the arterioles or venules in the connective tissue of host. The antigen recognized by IgM antibody of rats or mice was also observed in the parenchyme of sparganum and in the connective tissue of host. By 5∼20% gradient SDS-PAGE and EIBT, we detected antigenic components by IgG and 1gG antibodies of the rat or mouse immunized by infection or injection of crude extract of spargana. Twenty-three antigenic bands from crude extracts of spargana were recognized by IgG antibody and 15 components by IgM antibody of immunized rats. Out of the bands recognized by IgG and IgM antibodies, 15 were cross-reacted each other. Twenty components of eBlcretory-secretory proteins from spargana were recognized by IgG, and 5 components by IgM antibody of immunized rats. By IgG and IgM antibodies of immunized mice, 16 components of crude extracts were recognized by IgG antibody and 9 components by IgM antibody. Twenty components of excretory-secretory preparation were recognized by IgG antibody and 5 components by IgM antibody. Thirteen components of crude extracts were cross-reacted by IgG antibody of rats and mice.

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Seroepidemiological study of Neospora caninum in Korean dairy cattle by indirect immunofluorescent antibody assay (간접형광항체법을 이용한 국내 젖소의 Neospora caninum에 대한 혈청역학적 연구)

  • Hur, Kwon;Kim, Jae-hoon;Hwang, Woo-suk;Hwang, Eui-kyung;Jean, Young-hwa;Lee, Byung-chun;Bae, Ji-seon;Kang, Yung-bai;Yamane, Itsuro;Kim, Dae-yong
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.859-866
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    • 1998
  • This study was carried out to investigate the prevalence of Neopsora (N) caninum infection in Korean dairy herds. To determine the prevalence of antibodies to N caninum in Korean dairy cattle, a total of 1,688 sera including 895 sera taken from 30 herds having recent high abortion rate and 793 sera selected randomly from 168 herds with no history of recent abortion problem, respectively, collected nationwide during a designated period were analyzed by indirect fluorescent antibody (IFA) test. Mean nationwide seropositive rate of the sera tested in herds and individual cattle tested were 53.5% and 35.6%, respectively. However mean seropositive rate of the samples from herds having abortion problem was approximately two and half times higher than those in herds with no recent abortion history. Regional seropositive rates of the samples from the herds with abortion problem were 48.6%, 51.6%, 44.4% and 71.4% at Kyunggi, Kangwon, Kyungbuk and Jeonnam province, respetively. Regional seropositive rates of the samples from the herds with no recent abortion problem were 35.6%, 18.3%, 16.5%, 37.5%, 19.4%, 33.3%, 32.1%, 3.8% and 0.0% at Kyunggi, Kangwon, Chungbuk, Chungnam, Kyungbuk, Kyungnam, Jeonbuk, Jeonnam and Jeju province, respectively. The results of this study suggested that N caninum infection was widespread and considerably associated with bovine abortion in Korea.

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Comparative Analysis of Phospholipase D2 Localization in the Pancreatic Islet of Rat and Guinea Pig

  • Ryu, Gyeong-Ryul;Kim, Myung-Jun;Song, Chan-Hee;Min, Do-Sik;Rhie, Duck-Joo;Yoon, Shin-Hee;Hahn, Sang-June;Kim, Myung-Suk;Jo, Yang-Hyeok
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.4
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    • pp.211-215
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    • 2003
  • To examine the localization pattern of phospholipase D2 (PLD2) in the pancreatic islet (the islet of Langerhans) depending on species, we conducted a morphological experiment in the rat and guinea pig. Since individual islets display a typical topography with a central core of B cell mass and a peripheral boundary of A, D, and PP cells, double immunofluorescent staining with a panel of antibodies was performed to identify PLD2-immunoreactive cells in the islets PLD2 immunoreactivity was mainly present in A and PP cells of the rat pancreatic islets. And yet, in the guinea pig, PLD2 immunoreactivity was exclusively localized in A cells, and not in PP cells. These findings suggest a possibility that PLD2 is mainly located in A cells of rodent pancreatic islets, and that the existence of PLD2 in PP cells is not universal in all species. Based on these results, it is suggested that PLD2 may play a significant role in the function of A and/or PP cells via a PLD-mediated signaling pathway.

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
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    • v.15 no.3
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    • pp.207-210
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    • 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.

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Study on the Pathogenesis of Hantaan Virus with Monoclonal Antibodies (단일항체를 이용한 한국형출혈열의 병인성 연구)

  • Kim, Gum-Ryong;Kim, Tai-Gyu;Rhyu, Mun-Gan;Lim, Byung-Uk
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.1
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    • pp.1-8
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    • 1987
  • Hantaan virus(HV) 76-118 strain was inoculated into suckling ICR mice by intra-nasal route with an inoculum of $10LD_{50}$. Mortality was 65% at the 3rd week after inoculation, but declined to 35% at the 4th week. Infectivity was determined by the measuring immuno-fluorescent antibody in sera. The peak of infectivity was 80% at the 4'th week after inoculation. Viremia was reached peak level of $1.7{\times}10^4\;PFU/ml$ by day 10. Immunofluorescent antibody and neutralizing antibody appeared by 2 weeks and 15-17 days respectively, but achieved similar titer by 35 days. By using a monoclonal antibody to HV 76-118, viral antigens were initially detected in inguinal and axillary lymph node by 2 days. Viral antigens in bone marrow and lung were delayed much more than in those of lymph node. These were similar with those of intra-peritoneal and intra-muscular route. Immune complex against IgG, IgM and C3 appeared by 16 days, 14 days, and 18 days respectively. The pattern of immunofluorescence in the basement membrane of glomeruli was diffuse membranous. Spotted pattern was also observed in the tissue stained with anti-mouse C3 antibody. By 20 days, control tissue was also shown immune complex in the glomeruli.

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A Seroimmunologic Study of Bats Infected with Hantavirus in Korea ($1989{\sim}1995$) (한국 박쥐의 한타바이러스 감염에 대한 혈청면역학적 연구)

  • Park, Eun-Byung;Cho, Kyu-Bong;Park, Chul-Hee;Lee, Yun-Tai
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.91-99
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    • 1996
  • To understand whether the bats inhabiting in Korea play role as hosts harboring Hantavirus that cause acute febrile diseases, a total number of 802 bats of 9 species were collected from seven provinces in Korea from 1989 to 1995 and tested for the presence of antibodies to Hantavirus by means of immunofluorescent antibody (IFA) technique. The results are summarized as follow. 1. Total 802 captured bats were classified into 9 different species with the following distribution. They were Rhinolophus ferrumequinum, Eptesicus serotinus, Miniopterus sehreibersii, Vespertilio superans, Myotis mystatinus, Murina leucogaster, Myotis formosus, Myotis macrodactylus and Plecotus auritus with numbers and rates of 423 (52.74%), 291 (36.28%), 47 (5.86%), 28 (3.49%), 8 (1.00%), 1 (0.12%) and 1 (0.12%), respectively. The predominant species of the bats was Rhinolophus ferrumequinum with 52.74% of the captured. 2. Among 9 species of bats, species of Rhinolophus ferrumequinum and Eptesicus serotinus were positive with Hantavirus antibody of strain numbers 76-118. The rate of antibody positive was 3.78%. 3. The seasonal differences of Hantavirus antibody in 802 bats tested were 5.83%, 4.17%, 3.67% and 0.64% in winter, spring, summer and autumn, respectively. Again the highest viral antibody prevalence was detected in winter. It could be concluded through the study that certain species of bats inhabiting in Korea play a definite role as the host animals of certain species of Hantavirus.

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Development of competitive enzyme linked immunosorbent assay for detection of Coxiella burnetii antibody in animal (동물에서 Coxiella burnetii 항체를 진단하기 위한 경쟁효소면역법 개발)

  • Cho, Dong-hee;Kim, Yong-ju;Wee, Sung-hwan;Cho, Mi-young;Kweon, Chang-hee;Kang, Yung-bai;Park, Yong-ho;Cho, Sang-nae
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.81-85
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    • 2000
  • Coxiella burnetii (C burnetii) is the causative agent of Q fever in animal and human. The distribution of the disease has been documented around world. In this study we developed the competitive enzyme linked immunosorbent assay(cELISA) and compared it with indirect immunofluorescent assay(IFA). A monoclonal antibody(Mab) against C burnetii and a peroxidase-conjugated anti-mouse IgM were used as an indicator system competing against antibody in animal serum or as an indicater of the absence of antibody. Sera were considered antibody positive when the percentage inhibition index(PI index) is upper than 30. PI index is calculated as 100-[sample OD/Mab OD)${\times}100$]. Among 162 bovine serum samples, 23 samples were antibody positive both in cELISA and IFA. And 156 samples showed same results. From goat with experimentally induced infection with C burnetii the antibody was detected 20 days early in cELISA compared to IFA. On the basis of present findings, it was demonstrated that cELISA is a reliable diagnostic method for The detection of specific antibodies against C burnetii infection.

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Role of cytoskeleton in Host Cell Invasion by Intracellular Protozoa Toxoplasma gondii

  • Lee, Sook-Hwan;Lee, Boo-Young;Min, Duk-Young;Kim, Jung-Mogg;Ahn, Myoung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.628-634
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    • 2002
  • A microfilament-based motility in Toxoplasma gondii (T. gondii) Is involved in host cell invasion, yet the exact mechanism has not yet been determined. Accordingly, the current study examined the localization of actin and tubulin in T gondii using immunofluorescent (IF) and immunogold staining for electron microscopy. Indirect immunofluorescence (IF) staining using anti-actin and anti-tubulin monoclonal antibodies (mAbs) revealed localization of fluorescence on the entire surface of the tachyzoites. The actin in T. gondii was observed by immunogold staining, and the gold particles were seen on the surface, especially at the anterior end and in the cytoplasm of the parasite. However, there were no gold particles in the nucleus, rhoptries, and dense granules. The tubulin in T gondii was located on the surface and in the cytoplasm of the tachyzoites in the extracellular parasite, compared with anterior part of tachyzoites in the intracellular parasite. The antigens of T gondii recognized by anti-actin mAb were 107 kDa, 50 kDa, 48 kDa, and 40 kDa proteins, while those recognized by anti-tubulin mAb were 56 kDa, 52 kDa, and 34 kDa proteins. Tachyzoites of T gondii pretreated with the actin inhibitor, cytochalasin D (20 $\mu\textrm{g}$/ml), and tubulin inhibitor, colchicine (2$\times$10$\^$-6/ M), for 30 min at 37$\^{C}$ were used to infect the isolated mouse macrophages (tachyzo ites:macrophage=2:1). Pretreatment with the inhibitors resulted in lower multiplication of tachyzoites within the macrophages than in the untreated group 18 h post infection (p<0.05). Therefore, the present results suggest that actin and tubulin appear to be involved in the invasion of and multiplication in host cells.

Anti-epidermal growth factor receptor (EGFR) monoclonal antibody and DNA topoisomerase inhibitor reduce growth of salivary adenoid cystic carcinoma in a murine model (항-표피성장인자수용체 단클론항체와 DNA 토포이소머라제 억제제에 의한 마우스 모델에서의 타액선 선낭암종 성장 억제)

  • Park, Young-Wook;Lee, Hee-Su
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.3
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    • pp.177-185
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    • 2010
  • Introduction: Epidermal growth factor receptor (EGFR) is expressed in human epithelial tumors including salivary cancers, and known to be correlated with tumor progression and poor clinical courses in some epithelial tumors. In this study, we determined the therapeutic effect of the anti-EGFR monoclonal antibody Erbitux (C225, cetuximab) in combination with the DNA topoisomerase I inhibitor irinotecan (CPT-11) on human salivary adenoid cystic carcinoma (SACC) cells growing in nude mice. Materials and Methods: At first, immunocytochemical analysis for the expression of EGFR and phosphorylated EGFR (pEGFR) on a human salivary ACC cell line (ACC3). To determine the in vivo effects of Erbitux and CPT-11, nude mice with orthotopic parotid tumors were randomized to receive intraperitoneal Erbitux (1 mg) two times per week, intraperitoneal Irinotecan (50 mg/kg) once per week, Erbitux plus CPT-11, or placebo. (control) Tumor volume and weight were measured. And mechanisms of in vivo activity of Erbitux and/or CPT-11 were determined by immunohistochemical/ immunofluorescent analyses. Results: Immunocytochemical staining of ACC3 demonstrated that EGFR was expressed and phosphorylated. CPT-11 inhibited ACC tumor growth in nude mice. Tumors of mice treated with CPT-11 and CPT-11 plus Erbitux exhibited increased tumor cell apoptosis and decreased microvessel density, which correlated with a decrease in the tumor volume in nude mice. But, CPT-11 seems not to be synergistic with Erbitux in our ACC3 model system. Conclusion: These results suggest that anti-EGFR monoclonal antibody and the DNA topoisomerase I inhibitor will be effective in the treatment of recurred or metastatic lesions of salivary ACC.