• Title/Summary/Keyword: antibody response

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The Radio-Immunoassay Method for Ginsenoside $Rg_1$ of Korean Ginseng

  • Han, Byung-Hoon;Han, Yong-Nam;Sankawa, Ushio;Akyama, Minko;Kawashima, Koichiro
    • Korean Journal of Pharmacognosy
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    • v.11 no.3_4 s.43
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    • pp.133-140
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    • 1980
  • In order to develop the radio-immunoassay procedure for the ginsenoside $Rg_1$ we prepared the $Rg_1-BSA$ conjugate and $Rg_1-tyramine$ conjugate by condensing the $Rg_1-azide$, which was prepared by a series of six step chemical modification of the $Rg_1-side$ chain, with bovine serum albumin(BSA) or with tyramine. Rabbits were immunized by repeated injection of $Rg_1-BSA$ conjugate with Freund's Complete Adjuvant for 5 month long to obtain very potent $anti-Rg_1$ serum. The radio-labelled haptene was prepared by direct radio-iodination $(125_J)$ of $Rg_1-tyramine$ according to the chloramine-T method. The radio-immunoassay procedure was successfully furnished by using DCC method (dextran coated charcoal) and the anti-body titer of the anti-serum was found as being $1600{\sim}3200$ by using 15000cpm tracer per test. Calibration test using non-labelled $Rg_1$ showed linear competetive binding response in the $(8-300){\times}34pg$. range of non-labelled $Rg_1$. The cross reaction test using 19 ginsenoside analogues enabled us a full structure-activity analysis on the antigen-antibody reaction that the anti-body in the serum would recognize the full structure of ginsenoside $Rg_1$ except the side chain moiety.

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Comparison of Efficacy of Newcastle Disease Virus $B_1$ Vaccine by Different Administration Method in Commerical Layer Chickens (뉴캣슬병 $B_1$ 생독백신의 효능비교)

  • 정만호;강춘원;노용기;박진열
    • Korean Journal of Veterinary Service
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    • v.13 no.1
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    • pp.96-102
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    • 1990
  • The immune responses of commercial layer chickens against Newcastle disease(ND) were compared among different administration methods and times of vaccination during 4 weeks of age. A total of 372 day-old chickens were divided into 4 groups of 93 birds each. Each of 3 groups was received a commercially available B$_1$live vaccine via drinking water, eye instillation or spray method at one, 14 and 28 days of age. One group was used as an unvaccinated control. At two and 4 weeks after each time of vaccination, 15 birds from each group were collected randomly out and challenged with virulent ND virus at the dose of $10^5E1D_{50}$ per bird. Ten to 15 birds from each group were bled at two weeks intervals from day old to 8 weeks of age for hemagglutination inhibition antibody titer, The protection rate was generally low regardless of the times of vaccination although two or more times vaccination gave higher protection than once vaccination. The low protection was considered due to low titer of the vaccine used since the vaccine titer was less than $10^{3.5}EID_{50}$ per bird. Spray method gave better protection compared to eye instillation or drinking water method which resulted in lowest response. Majority of birds showed clinical signs of ND between 3 and 6 days after challenge. Death occured one or two days after onset of symptoms. Major clinical signs observed were depression(94%), anorexia(84%), diarrhoea(29%), difficult breath(15%) and torticollis(10%). Hemorrhagic lesions on post mortem were seen in duodenum(51%), trachea(35%), illeum(13%), ceacal tonsil(11%), proventriculus(10%) and some other odrgans.

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IgA response in mice infected with Neodiplostomum seoulensis (서울주걱흡충 감염 마우스의 IgA 반응)

  • Sun HUH;Soo-Ung LEE;Moo-Ho WON;Young-Gil JEONG;Young-Hyun KWON;Chang sig CHOI
    • Parasites, Hosts and Diseases
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    • v.33 no.1
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    • pp.55-60
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    • 1995
  • To observe the production of IgA in Balb/c mice with neodiplostomiasis, 20 mice were infected with each 200 metacercariae of Neoniplostomum seoulenis. Sera and the duodenums were obtained 3, 7, 14, 28 days post-infection (PI) from five mice each group Neodipeostomum specific IgA in serum by the enzyme-linked immunosorbent assay increased from 7 days PI and persisted till 28 days PI. Immunohistochemistry for lgA was done with sections of the duodenum. The IgA-positive reaction was generally seen in the lamina propria and submucosa. Some of epithelial cells were positive at 7 and 14 days PI. The present finding showed that Neodiplostomun specific IgA antibody increased in serum and that there was local reaction of IgA in the mucosa and submucosa of the duodenum but not directly related with worm expulsion.

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Effect of ganglioside GD3 synthase gene expression on VSMC proliferation via ERK1/2 pathway, cell cycle progression and MMP-9 expression

  • Lee, Young-Choon;Kim, Cheorl-Ho
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2004.10a
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    • pp.81-90
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    • 2004
  • Sialic acid containing glycosphingolipids (gangliosides) have been implicated in the regulation of various biological phenomena such as atherosclerosis. Recent report suggeststhat exogenously supplied disialoganglioside (GD3) serves a dual role in vascular smooth muscle cells (VSMC) proliferation and apoptosis. However, the role of the GD3 synthase gene in VSMC responses has not yet been elucidated. To determine whether a ganglioside is able to modulate VSMC growth. the effect of overexpression of the GD3 synthase gene on DNA synthesis was examined. The results show that the overexpression of this gene has a potent inhibitory effect on DNA synthesis and ERK phosphorylation in cultured VSMC in the presence of PDGF. The suppression of the GD3 synthase gene was correlated with the down-regulation of cyclinE/CDK2. the up-regulation of the CDK inhibitor p21 and blocking of the p27 inhibition,whereas up-regulation of p53 as the result of GD3 synthase gene expression was not observed. Consistently, blockade of GD3 function with anti-GD3 antibody reversed VSMC proliferation and cell cycle proteins. The expression of the CD3 synthase gene also led to the inhibition of TNF--induced matrix metalloproteinase-9 (MMP-9) expression in VSMC as determined by zymography and immunoblot. Furthermore, GD3 synthase gene expression strongly decreased MMP-9 promoteractivlty in response to TNF-. This inhibition was characterized by the down-regulation of MMP-9,which was Iranscriptionally regulated at NF-B and activation protein-1 (AP-1) sites in the MMP-9promoter Finally, the overexpression of MMP-9 in GD3 synthase transfectant cells rescued VSMC proliferation. However MMP-2 overexpression was not affected the cell proliferation. These findings suggest that the fl13 synthase gene represents a physiological modulator of VSMC responses that may contribute to plaque instability in atherosclerosis.

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Study on the immunogenicity of synthetic polypeptide vaccine derived from Theileria sergenti merozoite (Theileria sergenti merozoite의 합성 polypeptide 백신의 면역원성에 관한 연구)

  • Baek, Byeong-kirl;Seo, Chang-hee;Kim, Jin-ho;Kim, Byeong-su
    • Korean Journal of Veterinary Research
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    • v.35 no.1
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    • pp.87-94
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    • 1995
  • Western immunoblot analysis of antigen of T sergenti merozoite revealed that the immunodominant proteins of this organism were characterized as the 18KD, 29KD, 34KD, 45KD and 105KD in Korea. The 34KD and 45KD among those immunodominant proteins of the parasite were isolated and their amino acid sequences from the $NH_2$-terminus were determined and synthesized. They respective polypeptides were cationized to enhance their antigenicity, fortified with Freund's adjuvant and tested for immunogenicity in rabbits and cattle. The results obtained were as follows; 1. Theileria sergenti merozoite antigen was shown in 120KD, 100KD, 66KD, 45KD, 34KD and 30KD in western immunoblot using serum of rabbits immunized with 34KD synthetic polypeptide and 70KD, 58KD, 55KD and 45KD using bovine serum. In western immunoblot, 45KD, 34KD and 30KD were recognized by immunized rabbits, and 50KD and 45KD by cattle sera immunized with 45KD synthetic polypeptide, respectively. 2. The ELISA utilizing the synthetic polypeptides demonstrated significant antibody response to the respective peptides. After the 2nd booster injection, an OD of 0.760(preimmunization 0.132) in rabbits and an OD of 0.645(preimmunization 0.488) to 34KD synthetic polypeptide in cattle were observed. In animals immunized with 45KD synthetic polypeptide, after the 2nd booster injection, an OD of 0.640(preimmunization 0.144) in rabbit, and an OD of 0.776 (preimmunization 0.477) in cattle were measured. 3. After the 2nd booster the reciprocal IFA titer was 1:64 in rabbits and 1:512 in cattle immunized with the 34KD synthetic polypeptide. The IFA titre was observed as 1:512 in rabbit and 1:1,024 in cattle in immunized with the 45KD synthetic polypeptide.

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Enhancement of cell-mediated immunity by administration of plasma protein in pigs 1. Proportion of leukocyte subpopulations and cells expressing adhesion molecules in peripheral blood (돼지에서 plasma protein에 의한 세포성면역 증진효과에 관한 연구 1. 혈액내 백혈구 아군 및 세포별 분포율)

  • Yang, Chang-kun;Kim, Soon-jae;Moon, Jin-san;Jung, Suk-chan;Park, Yong-ho
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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    • pp.275-286
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    • 1994
  • Plasma protein which has been known as one of nonspecific immunostimulators was added to feedstuff to examine its effect on the enhancement of cellular immune response in porcine immune system. A total of 40 piglets, 20 male and 20 female each, were fed for 30 days with or without plasma protein. The peripheral blood were collected and analyzed for the investigation of leukocyte subpopulations and their activities by using a panel of monoclonal antibodies specific to porcine leukocyte differentiation antigens and flow cytometry. The results obtained as follows. 1. Total weight gain, daily feed intake and feed conversion rate for 10 days were significantly improved to 56%, 20% and 22% in the piglets fed plasma protein, respectively. 2. A significant increase in N (null or non T/non B) cells was also noticed. Leukocyte proportion from piglets fed plasma protein was 20.2-24.7%, otherwise that from piglets fed without plasma protein was 12.3-13.4%, respectively. 3. A significant increase in the proportion of B cells and cells expressing poCD1 was not found in piglets fed plasma protein. 4. Reaction with monoclonal antibodies specific to adhesion molecules, poCD11a, poCD11b, poCD44 and poCD45A and poCD45B, has shown that leukocyte subpopulation from piglets fed plasma protein did not significantly higher than that from piglets fed without plasma protein. 5. Total proportion of granulocytes and monocytes was about 50% in both group and the proportion after treated with Hypaque/Ficoll was 2.7% and 5.8% in each group, respectively.

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An Experimental Studies on the Anti-allergic Reaction, the Antipyretic Action, the Anti-inflammatory and the Analgesic Action of Yenhwagamchotang and Gamiyenhwagamchotang (銀花甘草湯과 加味方이 抗 알레르기, 解熱, 消炎, 鎭痛效果에 미치는 影響)

  • Choe, Gyu-Dong;Kim, Jung-Ho;Chae, Byeong-Yun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.6 no.1
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    • pp.15-29
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    • 1993
  • Yenhwagamchotang has been widely used in treatment of inflammatory disease which is based on Oriental Medical literatures. These studies were attempted experimental effects of Yenhwagamchotang(sample A), Yenhwagamchotang plus Forsythia koreana NAKAI.(sample B), and Yenhwa-gamchotang plus Taraxacum mongolicum HAND- MAZZ(sample C),on the Anti-allergic reaction, the antipyretic action,the anti-inflammatory and the analgesic action,in rats. THe results of the studies were as follow: 1. Vascular permeability responses to intradermal serotonin in rats were showed significant effect at all sample groups. 2. The homologous passive cutaneous anaphylaxis in rats provoked by the IgE-like antibody aganist egg white albumin showed the decreasing effect. 3. The delayed type hypersensitivity responses to picryl chloride in mice were showed significant effect at all sample groups. 4. The delayed type hypersensitivity response to sheep red blood cell in mice were showed significant effect at all sample groups. 5. In anti-pyretic effect by yeast method were showed significant effect at all sample groups. 6. The anti-inflammatory effect by carrageenin method were showed significant effect at all sample groups. 7. The analgesic action by acetic acid method in mice were showed significant effect at all sample groups. According to the above result, Yenhwagamchotang(sample A), Yenhwagamchotang plus Forsythia koreana NAKAI(sample B), AND yenhwagamchotang plus Taraxacum mongolicum HNAN- MAZZ.(sample C ) were concluded to have the anti-allergic reaction, the antipyretic action, the anti-inflammatory, the analgesic action.

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Zinc Enhances Neutrophil Extracellular Trap Formation of Porcine Peripheral Blood Polymorphonuclear Cells through Tumor Necrosis Factor-Alpha from Peripheral Blood Mononuclear Cells

  • Heo, Ju-Haeng;Kim, Hakhyun;Kang, Byeong-Teck;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.37 no.5
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    • pp.249-254
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    • 2020
  • Neutrophil extracellular trap (NET) formation is an immune response for the invasion of microbes. The purpose of this study is to examine the effect of zinc on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation of PMNs was measured by fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α in the culture supernatants from zinc-treated peripheral blood mononuclear cells (PBMCs) was measured by enzyme-linked immunosorbent assay (ELISA). Zinc itself did not have no effect on NET formation. However, the NET formation of PMNs was increased by culture supernatants from PBMCs treated with zinc. Also, the NET formation of PMNs was increased by recombinant porcine (rp) TNF-α. The production of TNF-α in PBMCs culture supernatants was shown to increase upon zinc treatments. These NET formations of PMNs increased by either culture supernatant from PBMCs treated with zinc or rpTNF-α were inhibited by treatment of anti-rpTNF-α polyclonal antibody (pAb). These results suggested that zinc has an immunostimulating effect on the NET formation of PMNs, which is mediated by TNF-α released from zinc-treated PBMCs. Therefore, zinc may play an important role for NET formation in the defense of porcine inflammatory diseases.

A survey of viruses associated with reproductive failure in boar semen in Korean artificial insemination centers (국내 인공수정센터의 웅돈에 대한 번식 관련 바이러스 조사)

  • Kim, Yeong-Hun;Chun, Bong-Su;Kim, Sung-Jae;Han, Jeong-Hee
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.111-116
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    • 2011
  • Artificial insemination (AI) of swine is a very useful reproductive tool and that offers convenience in the Korean swine industry. Since many viruses have been reported to be excreted through boar semen, we investigated the presence of antibodies and antigens against viruses causing reproductive failure in semen of boar in 349 semen samples collected from six Korean AI centers. Viral antigens were detected by polymerase chain reaction (PCR) or reverse transcription-PCR predominantly. The results was as follows. The major reproductive failure causing factor was porcine circovirus type 2 (PCV2), followed by porcine reproductive and respiratory syndrome virus (PRRSV) ($X^2$=166.64, P<0.001). PCV2 and PRRSV, Japanese encephalitis virus (JEV), encephalomyocarditis virus (EMCV) was detected in 73 samples (20.9%), 44 samples (12.6%), 4 samples (1.1%), 3 samples (0.9%), respectively and porcine parvovirus in one sample (0.3%) Classical swine fever virus (CSFV), bovine viral diarrhea virus and Aujeszky's disease virus (ADV) were not detected. Enzyme-linked immunosorbent assay was carried out in 111 serum samples from three AI centers. In most pigs, antibodies response was showed prominently in CSFV (105 sera, 94.6%) ($X^2$=82.580, P<0.001), followed by, in PRRSV (100 sera, 90.1%), PCV2 (92 sera, 90.1%), and PPV (8 sera, 82.9%). ADV antibody was not detected. Thus, the experimental results will be used for the base data, with respect to the state of viral stillbirth in general pig farms, as well as AI centers and breeding farms in Korea.

An in vitro vitellogenin bioassay for estrogenic substances in the Rhynchocypris oxycephalus (버들치(Rhynchocypris oxycephalus)에서 에스트로겐류 물질에 의한 vitellogenin의 유도 및 단일클론항체 제작)

  • Shin, Jung-A;Song, Jae-Young;Lim, Hee-Young;Chang, Min-Ho;Lee, Hye-Min;Chung, Kyu-Hoi;Lim, Yoon-Kyu;Yoon, Byoung-Su
    • Korean Journal of Veterinary Research
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    • v.46 no.1
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    • pp.35-41
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    • 2006
  • Vitellogenin (Vtg), a phospholipoglycoprotein precursor of egg yolk is synthesized and secreted from the liver in response to estrogens in female fish. Vtg is normally undetectable in the blood of male fish, but can be induced by exposure to chemicals possessing estrogenic activity. Thus, the presence of Vtg in blood of male fish can serve as a useful biomarker for assessing previous exposure to estrogenic compounds. In the present study, Vtg was abnormally expressed in Rhynchocypris oxycephalus using estradiol benzoate ($E_2$). As the result, it was found that the level of Vtg in blood from R. oxycephalus was increased by treated quantity of $E_2$ with dose-effect manner. Monoclonal antibodies were generated against Vtg of R. oxycephalus. The hybridoma were screened with an enzyme immunoassay for the production of specific anti-Vtg antibodies. Five positive cell lines with a high specificity were selected. Monoclonal antibodies against vtg of R. oxycephalus that was developed in this study, may be a useful bio-indicator for the detection of estrogenic contamination in the aquatic ecosystem.