• Journal of Microbiology and Biotechnology
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• 제23권1호
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• pp.69-75
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• 2013

Enrofloxacin is a fluoroquinolone antibiotic approved for the treatment of infections in animals. Because of the side effects to consumers of animal products, the maximum residue limits (MRLs) of enrofloxacin in animal tissues for consumption are regulated. In this study, a monoclonal antibody (mAb) against enrofloxacin was prepared and characterized for the development of a direct competitive enzyme-linked immunosorbent assay (ELISA). The obtained mAb, Enro44, was highly specific for enrofloxacin and had a 50% inhibition concentration ($IC_{50}$) of 1.99 ng/ml in a competitive ELISA, and the limit of detection (LOD) was 0.50 ng/ml. The cross-reactivity of the mAb with other quinolones and fluoroquinolones was lower than 0.01%. The subclass of the mAb Enro44 was identified as IgG1. The antigen (Ag)-captured direct competitive ELISA using the mAb Enro44 was tested on different spiked samples, including chicken muscle, cattle milk, and cattle urine, and the assay demonstrated recoveries of 82-112%, 80-125%, and 78-124%, respectively. Furthermore, the quantitation of enrofloxacin obtained from the ELISA and from high-performance liquid chromatography (HPLC) was in good agreement, with the linear regression coefficient between 0.933 and 1.056. The cDNAs encoding a heavy-chain Fd fragment (VH and CH1) and a light chain of the mAb Enro44 were cloned and sequenced. Taken together, the results obtained reveal a potential use of this mAb in an ELISA for the detection of enrofloxacin in food samples. The information of amino acid sequence of this mAb will be useful for further modification and production of the mAb in a bioreactor.

• 대한수의학회지
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• 제35권3호
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• pp.583-593
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• 1995

The present study was conducted to develop a toxoplasma latex agglutination test antigen(Test-MT) and evaluate the toxoplasma latex agglutination(LA) test using a newly-made "Test-MT kit" by comparing with the Toxo-MT kit(Eiken chemical co, Tokyo). Also, the specifity and sensitivity test were made by comparing with IFA test and IgG-ELISA. Tachyzoite suspensions of Toxoplasma gondii(RH strain) were ultracentrifuged for 30min at $60,000{\times}g(4^{\circ}C)$ and the supernatant was used as a water-lysate antigen. Polystyrene latex particles of $1.0{\mu}m$ in diameter(Polyscience co) were used for the preparation of sensitized latex-antigen supension(Test-MT). The frequency distribution of LA titers in Test-MT showed two peaks at <1:32 and 1:128. The borderline titer for positive test in Test-MT was determined to be 1:64. But the frequency distribution of LA tites in Toxo-MT showed two peaks at <1:16 and 1:64. The positive borderline was determined to be 1:32. Agreement of reactions between Test-MT and Toxo-MT kit by LA test was shown 92.5% in bovine sera and 97.0% in swine sera, respectively. From the results obtained here it was determined that the sensitized latex-antigen, Test-MT kit, for the microtiter agglutination test prepared as same as by the procedure described in the previous paper(Suh and Lee, 1993) was useful as a highly specific, sensitive and stable immunotiteration reagent for serodiagnosis of toxoplasma infection in animal sera.

• 동의생리병리학회지
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• 제19권2호
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• pp.380-388
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• 2005

In this study, the immunological effect of a traditional Korea herbal medicine, Sochungyong-tang (SCRT) that has been widely used for the treatment of various immunological disorders including allergic asthma in Korea, was examined in vitro. In our previous study demonstrated that SCRT decreases the expression of IL-4 mRNA, that plays pivotal role in Th2 cell development, while increases $IFN-{\gamma}{\tilde{a}}expression$, which is one of the key cytokines for Th1 lineage development in Th0 condition. That study strongly implies that SCRT can correct Th2 dominant condition directly affecting to the CD4+ T cell development. Present study designated to further evaluate the SCRT on helper T cell development by monitoring Th1/Th2 specific cytokine secretion patterns in artificially induced Th1 or Th2 polarized condition. The results demonstrated that Th2 cells were dramatically under-populated in Th2 driven condition with SCRT treatment, while Th1 cells were not altered in Th1 skewed condition. Furthermore, under Th2-skewed conditions the levels of and IL-4 were considerably decreased with SCRT treatment. However, the expression of GATA-3, a transcription factor that plays pivotal role in Th2 lineage programming, was not changed with SCRT, suggesting that the suppression of Th2 cell development by SCRT was not mediated by GATA-3. Present study implies that the effect on CD4+ T cell may be the one of key pharmacological effect point for treating IgE medicated allergic asthma by SCRT. These results also suggest that SCRT might be desirable agent for the correction of Th2 dominant pathological disorders.

• Parasites, Hosts and Diseases
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• 제49권3호
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• pp.235-243
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• 2011

In order to get a better understanding of the role of protease-activated receptor 2 (PAR2) in type 2 helper T (Th2) cell responses against Trichinella spiralis infection, we analyzed Th2 responses in T. spiralis-infected PAR2 knockout (KO) mice. The levels of the Th2 cell-secreted cytokines, IL-4, IL-5, and IL-13 were markedly reduced in the PAR2 KO mice as compared to the wild type mice following infection with T. spiralis. The serum levels of parasite-specific IgE increased significantly in the wild type mice as the result of T. spiralis infection, but this level was not significantly increased in PAR2 KO mice. The expression level of thymic stromal lymphopoietin, IL-25, and eotaxin gene (the genes were recently known as Th2 response initiators) of mouse intestinal epithelial cells were increased as the result of treatment with T. spiralis excretory-secretory proteins. However, the expression of these chemokine genes was inhibited by protease inhibitor treatments. In conclusion, PAR2 might involve in Th2 responses against T. spiralis infection.

• 대한미생물학회지
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• 제22권3호
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• pp.197-208
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• 1987

Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.

• 생명과학회지
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• 제20권1호
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• pp.147-152
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• 2010

규칙적인 운동이 면역력 증가에 도움을 주는 것이 일반적인 사실이지만 차고 건조한 환경에서의 격렬한 운동은 호흡기 질환의 일종인 운동 유발성 천식과 운동 유발성 알레르기 질환을 야기하고, 상기도 감염 등을 유발하기도 한다. 이는 수행되는 운동의 종류나 방법 등에 따라서 다르게 나타나며 운동 수행의 경험과 운동 환경 등에 따라서 상이하게 나타난다. 따라서 운동하는 환경이 야외에서 이루어지며 오존이나 먼지 등을 많이 접하면서 수행되는 운동과 동계 스포츠인 스키, 스노우보드, 스케이트 및 아이스하키와 같은 운동 수행으로 운동 시 폐가 차갑고 건조한 공기에 장시간 노출됨에 따라 운동 유발성 천식과 운동유발성 알레르기 반응이 초래된다. 이러한 반응은 음식 알레르기와도 함께 나타나는데, 음식 알레르기가 있는 사람은 운동 시 더욱 알레르기 반응이 증가하게 되고, 음식 알레르기가 나타나지 않은 사람도 특정 음식 섭취 후 격렬한 운동 시 운동 유발성 알레르기가 나타나기도 한다. 이는 일반적으로 소화된 음식물이 장으로 지나갈 때, 혈액의 IgA가 덜 소화된 단백질을 혈액으로 들어오는 것을 방어해 주지만, 결렬한 운동이나 스트레스 상황에서는 IgA가 감소하게 되고, 이러한 상황에서 음식 유발성 알레르기 질환이 나타나게 되는데, 운동 시 이러한 반응이 더욱 심화되며 운동 유발성 알레르기 아나플락시스로 나타나기도 한다. 운동 유발성 알레르기 질환은 운동의 방법, 종류 및 수행되는 운동시간 등에 따라 깊은 관련이 있으며, 임상적인 징후로는 심한 기침, 가슴의 답답함, 호흡 곤란, 쌕쌕거림의 현상, 피부두드러기 및 혈관부종, 심할 경우 혈관파괴 등의 현상이 나타나고, 돌연사의 원인이 되기도 한다. 격렬한 운동은 과 호흡을 유발시키고, 이로 인해 폐의 비만세포가 증가하게 되며, 증가된 비만세포에서 나오는 히스타민이 알레르기 반응을 유발한다. 운동 유발성 두드러기나 아나필락시스 진단에 있어 가장 중요한 사항은 가족력과 병력에 대한 사항이며, 이외에도 메타콜린 피부 반응검사 등이 있다. 치료는 일상생활에서 활동의 수준과 범위를 수정하는 하는 것이 핵심이며, 이를 위해서는 전문가를 통한 환자 교육이 매우 중요하며, 식후에는 시간을 두고 운동해야 한다. 그리고 운동전 알레르기를 유발하는 음식물의 섭취를 제한해야 한다. 본 연구자는 운동 알레르기와 관련된 인자를 크게 운동 시의 환경과 음식물로 구분하고, 여러 문헌을 고찰하여, 운동 시 일어날 수 있는 알레르기 반응을 미연에 방지하고, 운동을 수행하는 엘리트 운동선수 및 동호인, 운동을 지도하는 지도자, 스포츠 산업에 종사하는 사람들에게 기초자료를 제공하고자 한다.

• 한국축산식품학회지
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• 제24권3호
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• pp.283-292
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• 2004

본 연구는 흰쥐(S.D, ♂)에 각 처리에 따라 기초사료로 시판사료(T$_1$)를 무제한 급여하면서 시험사료로 한약 증탕액 (T$_2$), 오골계 증탕액(T$_3$), 오골계 교잡종 육에 Flavourzyme을 0.1%첨가하여 가수 분해시킨 후 한약제와 혼합하여 증탕시킨 증탕액(T$_4$)을 35일간 경구 투여한 후 흰쥐의 혈청에 대한 glucose및 hormones과 면역학적 변화에 미치는 영향을 조사 한 결과는 다음과 같았다. 비만하였을 때나 과도한 스트레스를 받았을 때 insulin 함량이 증가한다고 하였으나 효소 처리 오골계 교잡종 증탕액을 급여한 처리구에서 체중 증가가 일반사료를 급여한 처리구에 비해 유의적으로(p<0.05) 증가하였어도 insulin 함량이 일반사료 급여구보다 증가하지 않은 것으로 나타나 바람직한 생리현상을 나타냈다. Aldosterone은 스테로이드 계통의 호르몬으로 물의 생리적 밸런스와 혈압조절에 관여하는데 일반사료 급여구에 비해 한약 증탕액, 오골계 증탕액,효소 처리 오골계 증탕액 급여 구에서 낮은 함량을 나타내 고농도 단백 식이의 급여에도 불구하고 aldosterone수치의 저하로 혈압 저하의 효과를 나타 낸 것은 한약을 첨가시켜 고압으로 처리시킨 증탕액의 효과와 가수분해 효소에 의한 소화가 용이하도록 처리한 결과로 사료된다 Cortisol은 스트레스 측정에 감도가 높은 호르몬으로 알려져 있는데 일반사료급여구가 0.67 nmol/L인 반면 증탕액을 급여한 구에서는 0.40∼0.49 nmol/L으로 cortisol 농도가 감소하는 경향을 나타냈는데 이는 aldosterone에서와 같이 한약제와 동물성 단백질을 섭취함에 따른 효과로 사료된다. 흰쥐에 증탕액 급여가 면역반응에 미치는 영향에서 IL-4, IFN-v 및 anti-BSA IgG의 역가는 일반사료를 급여한 처리구에 비해 오골계 증탕액 및 효소 처리 오골계 교잡종 증탕액 급여구에서 spleen과 serum두에서 증가하는 경향을 나타내었는데 앞에서 언급했던 바와 같이 한약(십전대보탕)의 효과와 소화시키기 쉬운 고농도의 단백질 가수분해 물질의 다량 섭취에 의해 면역 활성이 증가되었을 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제14권8호
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• pp.1138-1143
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• 2001

A total of 96 piglets were weaned at 19 and 13 days in Exp. 1 and 2, respectively, and allotted to one of four diets: three with different spray dried plasmas (SPs) and one with hydrolysed casein (HC). SPs were from pigs (SPP), mixed origin (SMP), and mixed origin with standardized level of immunoglobulins (SMPIG). All the diets contained 1.7% total lysine, 25% of the test protein source, 45% corn starch, 15% lactose, 2% sucrose, 7% soybean oil. At d 4 and d 2 in Exp. 1 and 2, respectively, piglets were perorally challenged with $10^{10}$ CFU E. coli K88. Growth performance, immunity, and health condition were measured for 15 days and 14 days in Exp. 1 and 2, respectively. To investigate apparent ileal digestibility and nutrient deposition, all piglets were sacrificed at d 14 in Exp. 2. In 1. 3 piglets died in HC diet and 1 in SPP diet. HC diet showed higher mortality (p<0.01) than other diets. In Exp. 2, no clinical sign of infection was detected, no difference for the content of E. coli K88 was found in feces at 4 and 6 days after the infection, and no E. coli K88 was found in the jejunum at the end of experiment. In both experiments, feed intake was lower for HC diet and ADG was 96, 106, 122 and 155 for HC, SPP, SMP and SMPIG diet, respectively (HC vs others, p<0.05; SMPIG vs other SP, p<0.01). Heal apparent digestibility of nitrogen in sacrificed piglets was higher for HC diet (p<0.05). After the challenge, K88-specific titers in saliva (Exp. 1) and in plasma (Exp. 2) were reduced in SMP and SMPIG. The piglets positive to the adhesion of the used E. coli strain to the intestinal brush borders had a significantly reduced growth (p<0.01) and a higher K88-specific IgA titer in plasma, in comparison with negative ones. This effect was independent of the diet. The data show the relevance of spray dried plasma sources and particularly of SP with standardized level of immunoglobulins for the feeding of early-weaned at the risk of infection by enterotoxigenic bacteria.

• Journal of Microbiology
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• 제40권3호
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• pp.183-192
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• 2002

Cells infected With equine herpesvirus type-1 (EHV-1) Produced both infectious and non-infectious Virus-related particles. Compared to the whole virion, non-infectious particles termed L-particles were deter-mined to lack 150 kDa protein, commonly known as nucleocapsid protein. The potential of L-particles to induce immune responses was studied in mice and foals. Intranasal immunization with L-particles or whole virions induced poor IgG antibody responses in mice. Interestingly, despite the poor antibody response, the conferred immunity protected the host from challenge infections. This was indicated by a significant reduction in virus titers in line with recovery towards normal body weight. Subsequently, the test on the usefulness of L-particles as immunizing agents was extended to foals. Immunization of specific-pathogen-free (SPF) foals resulted in similar results. As determined by a complement-fixing-antibody test (CFT), foals seroconverted when they were immunized either with inactivated L-particles or whole virions via intramuscular (i.m.) injections. The presence of the antibody correlated with the degree of protection. Beyond day 1 post challenge infection (p.i.), there was no virus shedding in the nasal mucus of foals immunized with whole EHV-1 virions. Virus shedding was observed in foals Immunized with L-particles but limited to days 6 to 8 p.i. only. In contrast, extended vim shedding was observed in non-immunized foals and it was well beyond day 14 p.i. Viremia was not detected for more than four days except in non-immunized foals. Immunization in mice via intranasal (i.n.) conferred good protection. However, compared to the i.n. route, a greater degree of protection was obtained in foals following immunization via i.m. route. Despite variation in the degree of protection due to different routes of immunization in the two animal species, our results have established significant evidence that immunization with L-particles confers protection in the natural host. It is suggested that non-infectious L-particles should be used as immunizing agents for vaccination of horses against EHV-1 infection.

• The Korean Journal of Physiology and Pharmacology
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• 제1권4호
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• pp.445-456
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• 1997

We previously reported that some components of ginsenosides decreased mediator releases evoked by the activation of mast cells with specific antigen-antibody reactions. This study aimed to assess the effects of ginsenosides ($Rb_2$, Re) on the mechanism of histamine release in the mast cell activation. We partially purified guinea pig lung mast cells by using enzyme digestion, the rough and the discontinuous percoll density gradient method. Mast cells were sensitized with $IgG_1$ and challenged with ovalbumin (OA). Histamine was assayed by fluorometric analyzer, leukotrienes by radioimmunoassay. Phospholipase D (PLD) activity was assessed more directly by the production of $[^3H]phosphatidylbutanol$ (PBut) which was produced by PLD-mediated transphosphatidylation in the presence of butanol. The amount of 1,2- diacylglycerol (DAG) were measured by the $[^3H]DAG$ labeled with $[^3H]palmitic$ acid or $[^3H]myristic$ acid. Pretreatment of $Rb_2$ ($300\;{\mu}g$) significantly decreased histamine release by 60%, but Re ($300\;{\mu}g$) increased histamine release by 34%. Leukotrienes release in $Rb_2$ was decreased by 40%, Re was not affected in the leukotrienes release during mast cell activations. An increasing PLD activity during mast cell activation was decreased by the dose-dependent manner in the pretreatment of $Rb_2$, but Re pretreatment facilitated the increased PLD activity during mast cell activation. The amount of DAG produced by phospholipase C (PLC) activity was decreased by $Rb_2$ pretreatment, but Re pretreatment was not affected. The amount of mass DAG was decreased by $Rb_2$ and Re pretreatment during mast cell activation. The data suggest that $Rb_2$ purified from Korean Red Ginseng Radix inhibits the DAG which is produced by the activation of mast cells with antigen-antibody reactions via both phosphatidylinositide-PLC and phosphatidylcholine-PLD systems, and then followed by the inhibition of histamine release. However, Re increases histamine release by stimulation of DAG production, which is mediated by phosphatidylcholine-PLD system rather than by phosphatidylinositide-PLC system, but inhibits the mass DAG production. Thus, it could be inferred that other mechanisms play a role in the increase of histamine release during mast cell activation.