• Korean Journal of Veterinary Research
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• v.42 no.2
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• pp.209-217
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• 2002

This study was performed to produce monoclonal antibodies (mAb) specifically reacting with chicken leukocyte surface antigens. Popliteal lymph node cells of BALB/c mice previously immunized through foot-pad with peripheral blood mononuclear cells (PBMC) of chickens separated by Ficoll-Histopaque method. They were fused with P3X63Ag14 mouse myeloma cells. A total of 34 hybridomas secreted antibodies specifically binding to the PBMC. According to the reactivity patterns with PBMC, the mAbs were divided into 4 groups. Group 1 mAbs (IIB3, IIB10, IIE10) specifically reacted with non-adherent lymphocytes but not with adherent cells which were mainly composed of thrombocytes and monocytes in PBMC culture. These mAbs were reactive with 25-59% of thymus cells and 42-64% of spleen cells of chickens. They did not show any significant reactivity with cells in the bursa of Fabricius, T-cell (MDCC-MSB1) and B-cell (LSCC-1104B1) lines. These results indicate that Group I mAbs specifically reacted with T-lymphocyte subpopulation. Monoclonal antibodies in Group II (IC6, IG2-2 and IID9) showed specific reactivity with monocytes but not with thrombocytes or non-adherent cells in PBMC culture. These mAbs, though not reacted with the chicken macrophage cell line, HD11, also bound to macrophages of the spleen and lung in immunohistochemical staining. Five mAbs in Group III showed characteristics of binding to lymphocytes and monocytes, but not to thrombocytes. Twenty-three mAbs in Group IV showed specific reactivity to lymphocytes, monocytes, and thrombocytes. Two mAbs (IC3 and IE9) in Group IV reacted with most of PBMC.

• The Korean Journal of Food And Nutrition
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• v.19 no.2
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• pp.201-206
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• 2006

Numerous investigators have studied various activities of natural products and have found that they have not only nutritional effects, but also beneficial properties to cure various diseases and to maintain good health. Job's Tear(Yul-Moo) is a grass crop that has long been used in traditional medicine and as a nourishing food. Although its mechanism of action remains unclear, Job's Tear has been reported to exhibit anti-inflammatory, stomachic, anti-allergic, and anti-spastic effects and has been used in China for the treatment of warts, rheumatism, and neuralgia. Previous results in our laboratory demonstrated that the ethanol extract and the water extract of Job's Tear exerted an immune regulatory function on mice cells in vitro. The present study was performed to investigate the ex vivo effect of Job's Tear on immune function. Seven to eight weeks old mice(Balb/c) were fed chow diet ad libitum and water extract of Job's Tear was administered orally every other day for four weeks at two different concentrations(50 and 500mg/kg B.W.). Splenocytes proliferation with mitogen stimulation with Con A and LPS was enhanced at 50 mg/kg B.W. of Job's Tear compared to those of the control group. The results of this ex vivo study showed that proliferation of splenocytes and macrophage activation were seen in the mice orally administrated 50 mg/kg B.W. of Job's Tear water extracts. In conclusion, this study suggests that Job's Tear extracts may enhance immune function by regulating splenocyte proliferation and the cytokine prodution capacity of activated macrophages in mice.

• BMB Reports
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• v.31 no.1
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• pp.58-63
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• 1998

In this study, a mouse monoclonal antibody (mAb) against gp41(584-618), the immunodominant epitope protein, was generated. For this purpose, BALB/c mice were immunized with double branched multiple antigenic peptides derived from the HIV-1 gp41(584-618) sequence, and antibody-secreting hybridoma were produced by fusion of mice splenocytes with SP2/0 myeloma cells. One clone producing an antigen specific mAb, termed KI-41(isotype IgG1) was identified, whose specific reactivity against gp41(584-618) could be confirmed by ELISA and Western blot analysis. Epitope mapping revealed the recognition site of the mAb KI-41 to be located around the sequence RILAVERYLKDQQLLG, which comprises the N-terminal region within the immunized gp41(584-618) peptied. Since this mAb recognizes this specific epitope within the HIV-1 gp41 without any cross-reactivity to other immunodominant regions in the HIV-2 gp35, KI-41 will provide some alternative possibilities in further applications such as the development of indirect or competitive ELISA for specific antibody detection in HIV-1 infection or for other basic researches regarding the role and function of HIV-1 gp41.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.8
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• pp.1179-1187
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• 2001

In fish both humoral and cell mediated immune responses have been reported whereas antibodies recognizing specific cellular populations have not yet been developed except for ones recognizing surface Ig molecules on B lymphocytes. Our aim was to develop and characterize monoclonal antibodies (Mabs) specific for the immune-related cells. Mabs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously sensitized against Israeli carp (I. carp) kidney mononuclear cells. We obtained 44 Mabs positively reacting with I. carp kidney mononuclear cells and partially characterized 7 Mabs in the morphological and mitogen-based proliferative aspects. Fluorescence-activated cell sorter (FACS) analysis against I. carp kidney cells by using 7 different Mabs showed 80.3% for ICK 17-4, 65.1% for ICK 2-3, 64.1% for ICK 25-1, 67.5% for lCK 22-1, 70.8% for ICK 16-2, 76.8% for ICK 13-2, 79.7% for ICK II-I. Panning method was used for the isolation of Mabs specific mononuclear carp spleen cells followed by Wright's stain. The stained cell populations were identified as monocytes (ICK 17-4, ICK 2-3, ICK 25-1, ICK 22-1 and ICK 16-2), lymphocytes (ICK 11-1), and a mixed cell population of monocytes and lymphocytes (ICK 13-2). In cell proliferation assay, monocytes purified by ICK 17-4, 2-3 and 22-1 efficiently responded to Con A and PHA, while ones separated by ICK 25-1 did not react with any mitogens. Lymphocytes isolated by ICK 11-1, though it is not known whether they are T or B cells, were more responsive to Con A than PHA or LPS, suggesting that fish immune cells are somewhat different from mammalian cells in responding to mammalian T or B cell mitogens.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.167-170
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• 2004

Lactic acid bacteria (LABs) have been proposed as a potential oral allergy-therapeutic means of modulating immune phenotype expression in vivo, via promoting or reducing cytokine production. This study investigated the ability of LABs to suppress allergic response via modulating cytokine production in mice splenocytes. BALB/c mice were intraperitoneally primed with ovalbumin together with alum adjuvant to invoke antigen-specific Th1/Th2 cytokine-secreting cell populations in splenocytes. Spleen cells from mice fed with Lactobacillus confusus PL9001 (KCCM-10245), L. fermentum PL9005 (KCCM-10250), L. plantarum PL9011 (KCCM-10358), and Bifidobacterium infantis PL9506 (KCCM-10406) suppressed the levels of Th2 cell cytokines such as IL-4 and IL-5 during antigen sensitization. In addition, all mice fed with LABs induced secretion of Th1 cell cytokines such as IL-2 in splenocytes. These results suggested that LABs are anti-allergic agents, in view of their Th1/anti-Th2 immunoregulation.

• Korean Journal of Veterinary Research
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• v.44 no.2
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• pp.259-268
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• 2004

Porcine circovirus type 2 (PCV-2) has been associated with various disease in pigs worldwide including postweaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS). In this study, monoclonal antibodies (MAbs) against PCV were produced, characterized and applications of MAbs as diagnostic reagents were described. Spleen or lymph node cells from BALB/c mouse immunized respectively with PCV-1, PCV-2 or expressed PCV-2/ORF2 proteins in baculovirus were fused with SP2/0 myeloma cells using polyethylene glycol (PEG) and hybridoma cells producing PCV-1 or PCV-2-specific antibody were screened by an indirect immunofluorescence (IIF) test. A total of fifteen MAbs were produced against PCV. Six MAbs were PCV-1-specific and nine were PCV-2-specific. All PCV-1-specific MAbs reacted with only PCV-1 and all PCV-2-specific MAbs were reactive with only PCV-2 by IIF test. None of the MAbs was reactive with porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV), porcine rotavirus (PRV), and transmissible gastroenteritis virus (TGEV). Some PCV-2-specific MAbs recognized the PCV-2 infected porcine tissues by IIF or immunohistochemistry (IHC) assay. From this experiment, it was confirmed that MAbs produced in this study were PCV-specific and could be used as reliable diagnostic reagents for PCV-1/PCV-2 detection and differentiation.

• Journal of Life Science
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• v.9 no.1
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• pp.106-110
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• 1999

Lipopolysaccharide(LPS) from Vibrio vulnificus was purified, the fatty acid composition was analyzed, and Limulus gelation activity and lethal toxic activity were tested in order to investigate the cause of cytotoxicity by V. vutnificus. These results were compared to those of Escherichia coli LPS and Salmonella typhimurium LPS. LPS from V. vulnificus had a different fatty acid composition from those of E coli and S. typhimurium. The major fatty arid from each LPS was lauric acid for E. coli, rapric acid for S. typhimurium, and myristic acid for V. vulnificus. The Limulus gelation activities of three LPSs were the same(0.1ng/ml) and the lethal toxicity in BALB/c mouse of V vulnificus LPS was similar to those of E. coli LPS and S. typhimurium LPS. Such factor as exotoxin need to be considered to be the cause of cytotoxicity by V. vulnificus LPS.

• Journal of Food Hygiene and Safety
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• v.28 no.2
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• pp.95-98
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• 2013

Streptococcus suis (S. suis) is a major swine pathogen and an emerging zoonotic agent and is an increasing public health problem across Asia. The present study was undertaken to estimate the antibacterial effect of GR extract and therapeutic effect of GR extract against S. suis infection in mice. At the concentration of GR extract 2.5 mg/ml, the antibacterial effect was not shown on S. suis. However, the antibacterial effect against S. suis was observed at the concentration of GR extract 5.0 mg/ml. Oral administration of GR extract at the dose of 10 mg/kg showed a therapeutic effect for S. suis infected BALB/c mice. The mortality of GR extract-treated mice at the concentration of 5, 10 and 20 mg/kg was 80%, 70%, and 50% at 12 days, respectively, while that of untreated mice was 100% at 8 days after a lethal dose of S. suis infection. The results of our study strongly indicate that GR extract has potential as an effective for S. suis infection in mice.

• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.603-608
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• 2005

We recently isolated a novel probiotic strain, Lactobacillus fermentum PL9005 (KCCM-10250), from infant feces and showed that it had a potential immunoenhancing effect. In the present study, a safety assessment of the bacteria was performed using a BALB/c mouse model. Mice were administered with L. fermentum PL9005 daily for 28 days. There were no detectable changes in body weight, feed intake, or clinical signs, and no significant difference in hematological parameters or blood biochemistry between the L. fermentum PL9005-fed and control groups. Bacterial translocation was detected in the mesenteric lymph nodes, liver, and spleen of some mice with and without L. fermentum PL9005 feeding, however, the organisms were not related to ingestion of L. fermentum PL9005; this was confirmed by PCR using a species-specific primer. No gross lesions were detected in the liver, spleen, or intestine of L. fermentum PL9005-fed or control mice. Mucosal thickness in the ileum, cecum, and colon of L. fermentum PL9005-fed mice was not significantly different from that of corresponding organs in control mice. No inflammation or epithelial cell degeneration in the intestines was observed in any mice. These results indicate that ingestion of L. fermentum PL9005 is safe in mice and can be applied in the functional food market.

• Journal of Marine Bioscience and Biotechnology
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• v.1 no.4
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• pp.298-304
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• 2006

In this study, we establish a thin-layer chromatography (TLC) immunostaining method for detecting starfish gangliosides. A new monoclonal antibody (MAb) against AG-2, the major gangliosides molecular species of Acanthaster planci, was produced by fusing hybridoma with splenocytes immunized to liposomal AG-2. BALB/c male mice were injected with liposomal AG-2 antigen, and immunized. Their splenocytos were isolated and fused with hypoxanthine-aminopterine-thimidine (HAT)-sensitive mouse myeloma cells. Hybridomas producing MAb reactive to AG-2 were cloned using the limited dilution method. Established hybridomas were cultured in eRDF medium. Crude MAb produced from clone 8D4 was purified with a magnesium pyrophosphate column. Enzyme immunoassay and TLC immunostaining of AG-2 were performed using the purified MAb. Structurally related gangliosides did not cross-react with anti-AG-2 antibodies. The detection limit of TLC immunostaining was 50 ng of AG-2. The newly established immunostaining method was further developed for detecting AG-2 distribution and qualitative analysis in tissues and/or organs. Our results show that the majority of AG-2 is present in the stomach of male A. planci, while AG-2 is distributed not only in the stomach but also in the the pyloric caeca of female A. planci.