• Biomolecules & Therapeutics
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• v.5 no.4
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• pp.344-350
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• 1997

The purpose of the present study was to produce and characterize a monoclonal antibody against human ${\beta}_2$-adrenergic receptor. Male BALB/c mice were immunized with glutathione S-transferase (GST) fusion protein of the C-terminal portion of the human ${\beta}_2$-adrenergic receptor which was expressed in E.Coli. The immunized splenocytes were fused with myeloma SP2/0-Agl4 cells. The resulting hybridomas were screened for the production of a monoclonal antibody which can recognize human ${\beta}_2$-adrenergic receptor, and then subcloned by limiting dilution. The resulting monoclonal antibody was named as mAb$\beta$CO2. The mono-clonal antibody $\beta$CO2 was determined as IgM subtype and then purified by anti-mouse IgM-agarose affinity chromatography. The results of ELISA, Western blot, and immunocytochemistry showed that mAb$\beta$CO2 recognized human ${\beta}_2$-adrenergic receptor in the ${\beta}_2$-adrenergic receptor-GST fusion protein and human spider-moid carcinoma cell line A431 with highly specific immunoreactivity, The monoclonal antibody $\beta$CO2 may provide useful tools for the study of the $\beta$-adrenergic receptor of human and other species including rats.

• Korean Journal of Veterinary Service
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• v.33 no.2
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• pp.105-111
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• 2010

Rabies virus which belongs to the genus Lyssavirus of the family Rhabdoviridae is known as a highly neurotropic virus and causes fatal encephalitis accompanied by severe neurological symptoms in almost all mammals, including humans. In this study, monoclonal antibodies (MAbs) against rabies virus were produced, characterized and applications of MAbs as diagnostic reagents were assessed Spleen and inguinal lymph node cells from Balb/c mouse immunized with purified rabies virus were fused with SP2/O myeloma cells using polyethylene glycol (PEG) and hybridoma cells producing rabies virus-specific MAbs were screened by an indirect fluorescent antibody test. A total of ten MAbs were produced against rabies virus. The protein specificity and neutralizing activity of MAbs were determined by Western blot analysis and fluorescent antibody virus neutralization test, respectively. As a result, two MAbs, 5G3 and 6H4 had specificity for nucleoprotein (N protein) and two other MAbs, 5B1 and 5C1 had neutralizing activity for rabies virus. Some MAbs recognized the rabies virus-infected bovine brain stem cells by immunohistochemistry (IHC) assay. In conclusion, it was confirmed that MAbs produced in this study were rabies virusspecific and could be used as reliable diagnostic reagents for the detection of rabies virus.

• YAKHAK HOEJI
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• v.51 no.1
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• pp.44-50
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• 2007

Acute septic shock is one of inflammatory diseases mediated by pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$. In this study, we examined the pathological difference and mechanism of lipopolysaccharides isolated from E. coli (E-LPS) or S. abortus (S-LPS) on inducing acute septic shock in ICR mouse. All mice were died by intraperitoneal treatment of S-LPS with 0.75 mg/kg, whereas E-LPS treated with even 3 mg/kg only showed 30% of mice lethal, indicating that S-LPS may be more feasible in triggering a strong septic shock condition. The secretion pattern of TNF-${\alpha}$, a critical pro-inflammatory cytokine in septic shock condition, was also distinct between E-LPS- and S-LPS-treated groups. Thus, S-LPS strikingly increased serum level of TNF-${\alpha}$ (6 ng/ml) at 1 h, while E-LPS just displayed at 2 ng/ml level. However the interaction of S-LPS with LPS receptor toll like receptor (TLR)-4, was not stronger than that of E-LPS, according to experiments with macrophage cell line RAW264.7 cells. Thus, E-LPS rather than S-LPS strongly enhanced the production of TNF-${\alpha}$. Interestingly, S-LPS more strongly up-regulated splenocyte proliferation, compared to E-LPS group, whereas there was no difference between S- or E-LPS treated groups in proliferation of Balb/c- or C57BL/6-originated splenic lymphocytes. Therefore, our data suggest that S-LPS is a more active endotoxin and that the strong septic shock-inducing effect of S-LPS seems due to the enhancement of early TNF-${\alpha}$ production and S-LPS-sensitive lymphocyte proliferation.

• Journal of Dairy Science and Biotechnology
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• v.14 no.2
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• pp.229-245
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• 1996

1. CM-Sephadex C-50-120 column을 사용하여 HLF를 효과적으로 정제하였다. CM-Sepha-dex 50-120 column chromatography를 수행한 결과, HLF는 NaCl 350 mM-500 mM 사이에서 용출되었으며, 용출된 분획을 SDS-PAGE를 수행해서 단일 band를 확인하였다. Anti-HLF antibody를 이용한 Western blotting 결과 nitrocellulose paper 상에 단일 band 가 나타나므로 HLF 가 효과적으로 정제되었다는 것을 알 수 있었다. 2. Con A, PWM, PWA LPS 등의 자극원으로 단핵구와 대식세포를 자극한 다음 BLF를처리한 배 양액을 IL-1 bioassy한 결과는 Con A 33%, PMA 33%, PWM 15%, LPS 35% 이고, HLF로 처리하여 IL-1 bioassay를 한 결과는 Con A 15%, PMA 22%, PWM 10%, LPS 5%로 감소된 것으로 나타났다. 3. K-562 세포를 이용한 colony forming assay에서 BLF가 10 g/ml일 때는 30%, 30 g/ml일 때는 35%, HLF는 10 g/ml일 때는 5%,30 g/ml일 때는 30%의 저해를 나타냈다. 4. Lactoferrin의 면역증강효과를 알아보기 위하여 hapten인 VCR-BSA를 투여 한 후, 생성되는 항체생성능력을 ELISA로 비교하였다. 그 결과 HLF 및 BLF 투여군에서 대조군에 비하여 adjuvant 효과를 관찰할 수 있었다. 또한 이때 macrophage 수는 대조군에 비하여 HLF와 BLF를 투여한 군이 증가됨을 알 수 있었다. 이러한 결과로 미루어 볼 때 LF는 macrophage를 활성화 시켜서 항체 생성 능력을 증가시키는 효과가 있음을 관찰할 수 있었다. 5. Balb/c mouse의 thymus로부터 분리한 CD4- CD8- 세포를 BLF로 처리하여 24 시간 배양한 후 CB4$^-$ CD8$^-$ 세포의 분화를 측정한 결과, CD4$^-$를 CD4$^+$ 로 분화하였다. 그리고 HLF로 처리하여 24 시간 배양 후 CD4$^-$ CD8$^-$ 세포의 분화를 측정한 결과, CD4$^-$ CD8$^-$를 CD4$^+$ CD8$^+$ 로 분화하였다. 6. Lactoferrin이 T cell의 IL-2 production에 미치는 영향은 PMA 처리군, PMA+OKT3처리군, LF 단독 처리군 보다 PMA+OKT3+LF를 처리한 군이 IL-2 생성에 영향을 미쳤다. 7. Lfcin B의 단일크론항체에 의해 인식되어지는 Lfcin B의 항원결정기는 ‘QWR’로 밝혀졌다.

• Journal of radiological science and technology
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• v.42 no.2
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• pp.113-117
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• 2019

The purpose of this study was to investigate the FA value which can produce the best T2-weighted images by measuring the signal intensity and noise according to the FA value change in the brain image and the abdominal image of the mouse using micro-MRI. Brain imaging and abdominal imaging of BALB / C mice weighing 20g were performed using 4.7T (Bruker BioSpin MRI GmbH) micro-MRI equipment, Turbo RARE-T2 (spin echo-T2) images were scanned at TR 3500 msec and TE 36 msec. The changes of the FA values were $60^{\circ}$, $80^{\circ}$, $100^{\circ}$, $120^{\circ}$, $140^{\circ}$, $160^{\circ}$ and $180^{\circ}$. We measured signal intensity according to FA values of ventricle and thalamus in brain imaging, The signal intensity of kidney and muscle around the kidney was measured in abdominal images. To obtain SNR and CNR, we measured the background signals of two different parts, not the tissue. In the brain (thalamus) image, the signal intensity of FA $100^{\circ}$ was 7,433 and SNR (6.49) was the highest. In the abdominal (kidney) image, the signal intensity was highest at 16,523 when FA was $120^{\circ}$, and the highest SNR was 8.54 when FA was $140^{\circ}$. The CNR value of the brain image was 1.38 at FA $60^{\circ}$ and gradually increased to 8.29 at FA $180^{\circ}$. The CNR value of the muscle adjacent to the kidney gradually increased from 2.36 when the FA value was $60^{\circ}$ and the highest value was 4,57 at the FA value $180^{\circ}$.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.6
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• pp.1453-1462
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• 2003

In order to study the effect of oral administration of Mawhangyounpye-tang against to asthma, astham was induced to allergy-sensitive Balb/c mouse with ovalbumin using method of Hatfield et al (1997). The changes of diameter lumen of upper portion of the trachea, lung weight, gross appearance of lung, histological profiles of lung and trachea, numbers of cellular compartments in the bronchoalveolar lavage fluid (BALF), numbers and morphology of the mast cells in the trachea, numbers of mucus-secretory cell in the broncus, morphology of the bronchus, ultramicroscopical appearance of surface of trachea and number of cilia and mucous-secretory cells by scanning electron microscope. Obtained results were as follows. 1. The diameters of trachea lumen were significantly decreased in asthma induced control groups and these decreasing were result from hypertrophy of mucous membrane. However, these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 2. Lung weights and black spots, which were result from infiltration of inflammatory cells, were significantly increased in asthma induced control groups but these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 3. Hypertrophy of mucous membrane of trachea and bronchus and !bronchioles in the lung, peritracheal, peribronchus and peribronchiolar inflammatory cell infiltration, and mucoid exudate deposit in the lumen were observed in asthma induced control groups but these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 4. Cellular compartments including neutrophil and eosinophil were dramatically increased in the BALF of asthma induced control groups but these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 5. Mast cell degranulation and decreasing of the numbers of mast cells were detected in the trachea of asthma induced control groups. However, these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 6. Shed, decreasing of cilia cell and increasing of mucous-secretory cells in the surface of the trachea of asthma induced control groups but these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. In conclusion, it Is considered that Mawhangyounpye-tang has somewhat favorable effect on the asthma because the asthma specific series of abnormalities in respiratory system were decreased after oral administratin of Mawhangyounpye-tang in this study. In future, it is needed that the toxicological and dosagespecific study of Mawhangyounpye-tang to use against asthma with safe.

• Parasites, Hosts and Diseases
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• v.35 no.3
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• pp.189-196
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• 1997

The purpose of this study is to determine whether nitric oxide is involved in the extracellular killing of Trichomoncs uasinalis by mouse (BALB/c) peritoneal macrophages and RAW264.7 cells activated with LPS or rIFN-γ and also to observe the effects of various chemicals which affect the production of reactive nitrogen intermediates (RNl) in the cytotoxicity against T. vnginnlis. The cytotoxicity was measured by counting the release of (3H)-thymidine from labelled protozoa and NOa was assayed by Griess reaction. Nemonomethyl-L-arginine (L-NMHA), Nenitro-L-arginine methyl ester (NAME) and arginase inhibited cytotoxicity to T. vaginnlis and nitrite production by activated mouse perioneal macrophagrs and RAW 264.7 cells. The addition of excess L-arginine competitively restored trichomonacidal activity of macrophages. Exogenous addition of FeSO4 inhibited cytotoxicity to T. vaginaLis and nitric products of macrophages. From above results, it is assumed that nitric oxide plays an important role in the host defense mechanism of macrophages against T ucfinalis.

• KSBB Journal
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• v.30 no.4
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• pp.148-154
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• 2015

Onion (Allium cepa) is one of the flavonoids-rich materials in human diet and onion peel, which is the onion by-products, contains over 20 times more quercetin than the flesh. In this study, to examine the anti-inflammatory effects of onion peel hot water extract (OPHWE), the cell viability, nitric oxide (NO), pro-inflammatory cytokines, such as interluekin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and IL-$1{\beta}$, were measured using the murine macrophage cell line RAW 264.7 cells. The Balb/c mice were used for an in vivo acute toxicity test and ICR mice were used for measurement of inhibition effects of croton oil-induced mouse ear edema. As a result, NO levels decreased in a dose-dependent manner. The production of IL-6, TNF-${\alpha}$, and IL-$1{\beta}$ was suppressed by 38%, 41%, and 34% respectively, compared with that of the LPS only group, without any cytotoxicity. The edema formation in the ICR mouse ear was also reduced compared to that in control. Moreover, there were no mortalities occurred in mice administered 5,000 mg/kg body weight of OPHWE. These results suggest that OPHWE has considerable anti-inflammatory activities and can be regarded as a potent candidate material to treat inflammatory diseases.

• Archives of Plastic Surgery
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• v.38 no.4
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• pp.339-344
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• 2011

Purpose: The goal of cancer surgery is complete removal of cancer tissue and prevention of recurrence. Surgeons can change the surgical instruments after total resection of the cancer mass. The purpose of this procedure is to prevent dissemination of the cancer cells attached to the surgical instruments. Authors hypothesize the possibility of local recurrence caused by the cancer cells attached to the surgical instruments in the skin cancer cases. Methods: Skin cancers were induced by using DMBA-TPA two-stage carcinogenesis model in 10 of Balb/c mice. In 2-weeks, skin cancer was developed in all 10 mice. cancer cell attached surgical instruments were made by pinching the removed cancer tissue using Adson tissue forcep 10, 20, 30 times each. To count number of cancer cells in each forcep with different number of pinching was done, the forceps were washed in 30 mL of the normal saline and Cytospin preparation was done. To make recurrence models from cancer cell attached surgical instrument, three incisions were made in normal skin of each mouse, and local seeding was done by pinching subcutaneous tissue in 10, 20, 30 times each by using Adson teeth forceps mentioned above as cancer cell attached surgical instrument. Results: All skin cancers were squamous cell carcinoma. Local recurrences were developed in 7 mice (3 in 10 times forceping site, 2 in 20 times forceping and 3 in 30 times forceping). In the cytospin test, the mean number of squamous cells in 100 microscope was 28.6 in 10 times, 47.2 in 20 times, 93.6 in 30 times, respectively. P value was 0.002 in Wilcoxon-Sign test. Conclusion: The number of cell count was significantly increased as number of pinching was increased. And these cells are able to induce local recurrence by local seeding. Considering this result, authors are able to confirm that the minimal handling in cancer surgery is important factor to prevent local recurrence.

• IMMUNE NETWORK
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• v.1 no.2
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• pp.162-169
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• 2001

Background: Nitric oxide (NO), a cytotoxic molecule is produced in various tissues including tumor cells during interleukin-2 (IL-2) therapy . Lymphokine-activated killer (LAK) cells are induced during IL-2 therapy, and have cytotoxic activity against tumor cells. The current study investigated the effects of NO synthesized in target cells or exposure of target cells to NO on the sensitivity of target cells to LAK cell cytotoxicity. Methods: Cytotoxicity was measured using 4 h chromium release assays. LAK cells which were induced by a 4 day incubation of BALB/c mouse splenocytes with IL-2 (6,000 IU/mL) were employed as effector cells. RD-995 skin tumor cells originated from a C3H/HeN mouse were employed as target cells. NO synthesis in target cells was induced by a 24 h incubation of RD-995 cells with $IFN{\gamma}$ (25 U/mL), TNF (50 U/mL) and IL-1 (20 U/mL). S-nitrosyl acetylpenicillamine (SNAP), an NO donor, was used to expose target cells to NO. $N^G$-monomethyl-L-arginine (MLA) and carboxy-PTIO were added during cytotoxicity assays to inhibit NO synthesis, and to scavenge NO produced by target cells, respectively. Results: Sensitivity of NO-producing RD-995 cells to LAK cell cytotoxicity was decreased by addition of MLA and carboxy-PTIO during cytotoxicity assays. However, the two reagents had no effect on the sensitivity of non-NO-producing RD-995 cells. Pretreatment of RD-995 target cells with SNAP increased the sensitivity in comparison with untreated cells. Conclusions: Sensitivity of target cells to LAK cell cytotoxicity is increased by target cell NO synthesis or exposure to NO. Further studies are needed to evaluate whether these in vitro results have relevance to in vivo phenomena.