• YAKHAK HOEJI
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• v.54 no.6
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• pp.494-499
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• 2010

We have been focussing on discovery of natural compounds that have immunoregulatory activities for many years. In the present study, we investigated if chlorogenic acid (CRA), a polyphenolic compound, has an immunoadjuvant activity. Prior to examining the immunoadjuvant activity, effect of CRA on proliferation of T- or B-lymphocyte was determined. Results showed that CRA enhanced the proliferation of those lymphocytes in dose-dependant manner (P<0.05), and the proliferation enhancement by CRA was appeared to be more effective to B-cells than to T-cells. Based on these observations, it was tested with bovine serum albumin (BSA) and Candida albicans cell wall (CACW) as antigenic sources if CRA has an immunoadjuvant activity. In experiments, BSA alone or a mixture of BSA plus CRA was injected intraperitoneally to mice (BALB/c strain). For a negative control, mice were given only diluent (DPBS) by the same route. In other experiment, CACW was tested by the same way as did with BSA. Three weeks after the first immunization these animals were boosted. Antisera collected from the mice one week after the booster were analyzed by ELISA. Results displayed that the induction of anti-BSA antibody was increased in mice that received the mixture of BSA and CRA as compared to anti-BSA induction in BSA only-given mice groups (P<0.05). In case of CACW, a similar observation as did with BSA was made, resulting in that there was app. 40% increased production of the anti-CACW antiserum from the combination (CACW plus CRA)-received mice as compared to antiserum induction from CACW alone-given animals. Taken all together, these data indicate that CRA has an ability of enhancing antibody production regardless of nature of antigenic sources. Presumably, activation of B-cell proliferation by CRA may plays an important role in the immunoadjuvant activity of the polyphenolic compound.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.5741-5745
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• 2013

Background: Heat-shock protein70 (HSP70) are intracellular protein chaperones, with emerging evidence of their association with various diseases. We have previously reported significantly elevated plasma-HSP70 (pHSP70) in pancreatic cancer. Current methods of pHSP70 isolation are ELISA-based which lack specificity due to cross-reactivity by similarities in the amino-acid sequence in regions of the protein backbone resulting in overestimated HSP70 value. Materials and Methods: This study was undertaken to develop a methodology to capture all isoforms of pHSP70, while further defining their tyrosine and serine phosphorylation status. Results: The methodology included gel electrophoresis on centrifuged supernatant obtained from plasma incubated with HSP70 antibody-coupled beads. After blocking non-specific binding sites, blots were immunostained with monoclonal-antibody specific for human-HSP70, phosphoserine and phosphotyrosine. Conclusions: Our novel immunocapture approach has distinct advantages over the commercially available methods of pHSP70 quantification by allowing isolation of molecular aggregates of HSP70 with additional ability to precisely distinguish phosphorylation state of HSP70 molecules at serine and tyrosine residues.

• Parasites, Hosts and Diseases
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• v.53 no.4
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• pp.493-496
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• 2015

Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis with worldwide distribution. The present study investigated the prevalence of T. gondii in dogs in Zhanjiang city, southern China, using both serological and molecular detection. A total of 364 serum samples and 432 liver tissue samples were collected from the slaughter house between December 2012 and January 2013 and were examined for T. gondii IgG antibody by ELISA and T. gondii DNA by semi-nested PCR based on B1 gene, respectively. The overall seroprevalence of T. gondii IgG antibody was 51.9%, and T. gondii DNA was detected in 37 of 432 (8.6%) liver tissue samples. These positive DNA samples were analyzed by PCR-RFLP at 3'- and 5'-SAG2. Only 8 samples gave the PCR-RFLP data, and they were all classified as type I, which may suggest that the T. gondii isolates from dogs in Zhanjiang city may represent type I or type I variant. This study revealed the high prevalence of T. gondii infection in dogs in Zhanjiang city, southern China. Integrated measures should be taken to prevent and control toxoplasmosis in dogs in this area for public health concern.

• Parasites, Hosts and Diseases
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• v.47 no.2
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• pp.125-130
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• 2009

To figure out the epidemiological status and relevance with other diseases in toxoplasmosis, we checked serum IgG antibody titers of 1,265 patients and medical records of seropositive patients. Seropositive rates were 6.6% by latex agglutination test (LAT) and 6.7% by ELISA. No significant differences were detected between sexes and age groups. The peak seroprevalence was detected in the 40-49-year-old age group. According to clinical department, Toxoplasma-positive rates were high in patients in psychiatry, ophthalmology, health management, emergency medicine, and thoracic surgery. Major coincidental diseases in seropositive cases were malignant neoplasms, diabetes mellitus, arthritis, chronic hepatitis B, chronic renal diseases, schizophrenia, and acute lymphadenitis, in the order of frequency. In particular, some patients with chronic hepatitis B and malignant neoplasms had high antibody titers. These results revealed that the seroprevalence of toxoplasmosis in a general hospital-based study was similar to that in a community-based study, and T. gondii seropositivity may be associated with neoplasms, diabetes, and other chronic infections.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.197-202
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• 2001

Although several methods have been developed and applied to control swine respiratory diseases, the disease induces severe economic impact to swine industry worldwide. As one of the new trials, application of egg yolk antibody(IgY) was attempted for the purposes and immune response in sera and egg yolk was analysed with ELISA in previous study. In this study, immunological specificity of the IgY was analysed by Western blot analysis. In the analysis of causative agents of atrophic rhinits, B bronchiseptica and P multocida 4D, proteins of 33, 40, 43, 67 and 141 kDa were specifically reacted with IgY Also, 40 and 110 kDa proteins were identified as the major immunogens in P multocida 3A. In A pleuropneumoniae serotypes 2 and 5, 40 kDa and 47 kDa proteins were found to be the major reactive ones. These results suggested that egg yolk antibodies from immunized hens was specific with antigens injected into hens and partially purified antigens, outer membrane proteins and dermonecrotic toxin, were more effective than bacterin for the production of specific antibody.

• Food Science of Animal Resources
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• v.33 no.2
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• pp.198-204
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• 2013

This study aimed to apply the Parallux system to detect (fluoro)quinone antibiotics residues in raw bovine milk. The immunogen enabled the generation of a specific antiserum with a titer of 1/40,000. The $Parallax^{TM}$ kit using the antibody displayed $IC_{50}$ value of 10 to 150 ppb for (fluoro)quinolone antibiotics. $Parallax^{TM}$ kit was also sensitive for the detection of incurred (fluoro)quinolone at Korean Maximum Residual Levels in raw bovine milk as the result of dose response test. Cross reactivities of the antibody with the common (fluoro)quinolones were determined to be norfloxacin, 100%; enrofloxacin, 100%; ciprofloxacin, 100%; danofloxacin, 100%; nalidixic acid, 40%. Lower detection limit (LOD) values of the $Parallax^{TM}$ kit in raw bovine milk were determined to be norfloxacin, 4 ppb; enrofloxacin, 5 ppb; danofloxacin, 5 ppb; ciprofloxacin, 5 ppb and nalidixic acid, 10 ppb. The $Parallax^{TM}$ kit was run 8 times with five different concentrations of norfloxacin to determine the coefficient of variation (CV, %) of intra-assay, which was between 2.7% and 11.8%. To confirm the precision among kit batches for the inter-assay, five different batch kits were tested with 2 different concentration of norfloxacin. The CVs of the inter assay were 4.2% at 50 ppb, and 7.2% at 10 ppb norfloxacin, respectively.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1336-1344
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• 2017

Hepatitis B virus (HBV) is a major cause of liver cirrhosis and hepatocellular carcinoma. With recent identification of HBV receptor, inhibition of virus entry has become a promising concept in the development of new antiviral drugs. To date, 10 HBV genotypes (A-J) have been defined. We previously generated two murine anti-preS1 monoclonal antibodies (mAbs), KR359 and KR127, that recognize amino acids (aa) 19-26 and 37-45, respectively, in the receptor binding site (aa 13-58, genotype C). Each mAb exhibited virus neutralizing activity in vitro, and a humanized version of KR127 effectively neutralized HBV infection in chimpanzees. In the present study, we constructed a humanized version (HzKR359-1) of KR359 whose antigen binding activity is 4.4-fold higher than that of KR359, as assessed by competitive ELISA, and produced recombinant preS1 antigens (aa 1-60) of different genotypes to investigate the binding capacities of HzKR359-1 and a humanized version (HzKR127-3.2) of KR127 to the 10 HBV genotypes. The results indicate that HzKR359-1 can bind to five genotypes (A, B, C, H, and J), and HzKR127-3.2 can also bind to five genotypes (A, C, D, G, and I). The combination of these two antibodies can bind to eight genotypes (A-D, G-J), and to genotype C additively. Considering that genotypes A-D are common, whereas genotypes E and F are occasionally represented in small patient population, the combination of these two antibodies might block the entry of most virus genotypes and thus broadly neutralize HBV infection.

• Korean Journal of Veterinary Service
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• v.31 no.1
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• pp.101-111
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• 2008

The purpose of this study was to investigate the protective effects against porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) in suckling piglets by oral administration of IgY. Twenty piglets were divided into two groups with the same number: group I (treated with IgY) and group II (not treated). Group I was administerd orally with IgY for three days from one-day-old and experimentally challenged with PEDV and TGEV at four-day-old. The other was administered with saline solution and challenged with PEDV and TGEV at four-day-old. Serum antibody titers against PEDV and TGEV were examined by enzyme-linked immunosorbent assay (ELISA) and the detection of PEDV or TGEV antigen from feces and small intestines was performed by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunofluorescence (IFA). The antibody titers of the group I was higher than that of the other, and lasted at the end of experiment. In the detection tests of both virus from feces and small intestine, the rate of the group I was lower. Based on these results, oral administration of IgY may be effective to prevent the diarrhea caused by PEDV and TGEV.

• Korean Journal of Veterinary Service
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• v.34 no.4
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• pp.297-301
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• 2011

Rabies virus is transmitted most commonly through a bite from an infected dog. Especially, stray dogs that are not excluded from contact with rabid wild animals can become rabies vectors. Therefore, serological survey of rabies virus from stray dogs in Seoul Metropolitan City was carried out in this study. To investigate prevalence of rabies antibodies in the stray dogs, serum samples were taken from 500 stray dogs between April and December 2010. Antibodies to rabies virus were detected by indirect ELISA. Of 500 tested sera, 147 (29.4%) were positive to rabies virus. Prevalence rates of rabies antibodies(PRRA) in northern and southern Han river region of Seoul were 26.4% and 33.2% respectively. PRRA in male and female dogs were 33.6% and 26.1% respectively. PRRA in less than 1 year, 1~2< years, 2~3< years, 3~5< years, 5~10< years and over 10 years old dogs were 12.7%, 21.6%, 26.4%, 36.4%, 32.5% and 46.4%, respectively. These results indicates that antibody seroprevalence to rabies is still not enough to prevent rabies and rabies vaccination is required to enhance the seroprevalence in the dogs. To improve the situation, measures to raise public awareness of rabies and its prevention is needed. Also, reducing stray dogs and keeping companion animals from contact with wild animals are indispensable for the prevention of rabies.

• Journal of Animal Science and Technology
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• v.47 no.1
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• pp.19-28
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• 2005

The objectives of the current study were to develop polyclonal antibodies in sheep against adipocyte plasma membrane(APM) proteins isolated from swine, to investigate tissue specificity, and to determine cytotoxic effects of antiserum on swine adipocytes. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver, and spleen were isolated using a 32% sucrose gradient. Adult male sheep was immunized three times at three week interval with the purified swine APM proteins. Antiserum was taken from immunized sheep at 10, 12, and 14 days after the third immunization. Antiserum expressed strong reactivity with APM proteins determined by enzyme-linked immunosorbent assay(ELISA), and the reactivity could be detected at dilutions in excess of 1 : 81,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver, or spleen. Tissue specificity of the antiserum was reconfirmed by Western immunoblotting using anti-sheep immunoglobulin G•alkalinephosphatase conjugate as a secondary antibody. The reactivity of antiserum to the external surface of fixed swine adipocytes was confmned by an immunohistochemical technique using anti-sheep immunoglobulin G-FITC. Confluent swine adipocytes in culture were lysed by antiserum treatment and cytosolie lactate dehydrogenase(LDH) was released as a dose-dependent patterns while adipocytes treated with normal sheep serum maintained their integrity and expressed low level of LDH. These results implicate that fat contents in the pigs can be reduced by immunological methods.