• IMMUNE NETWORK
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• 제14권3호
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• pp.164-170
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• 2014

JL1, a specific epitope on CD43, is a potential biomarker for the diagnosis of acute leukemia. Although qualitative assays for detecting leukemia-specific CD43 exist, there is a need to develop quantitative assays for the same. Here, we developed two novel monoclonal antibodies (mAbs), 2C8 and 8E10, recognizing different epitopes on CD43. These clones are capable of pairing with YG5, another mAb against JL1 epitope, because they were selectively obtained using sandwich ELISA. Antigens recognized by 2C8 and 8E10 were confirmed as CD43 by western blotting using the CD43-hFC recombinant protein. When expression on various leukemic cell lines was investigated, 2C8 and 8E10 displayed a disparity in the distribution of the epitope. Enzyme assays revealed that these mAbs recognized a sialic acid-dependent epitope on CD43. Using normal thymus and lymph node paraffin-embedded tissues, we confirmed a difference in the epitopes recognized by the two mAbs that was predicted based on the maturity of the cells in the tissue. In summary, we developed and characterized two mAbs, 2C8 and 8E10, which can be used with YG5 in a sandwich ELISA for detecting leukemia-specific CD43.

• 대한한방내과학회지
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• 제44권3호
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• pp.294-312
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• 2023

Objective: The purpose of this study is to evaluate the effects of GGX on an ovalbumin (OVA)-induced asthma mice model. Methods: Balb/c mice were challenged with OVA and then treated with three concentrations of GGX (100, 200, and 400 mg/kg). After sacrifice, the bronchoalveolar lavage fluid (BALF) or lungs of the mice were analyzed by fluorescence-activated cell sorting, ELISA, real-time PCR, H&E, Masson's trichrome, PAS and AB-PAS staining, and immunohistofluorescence staining. Results: GGX significantly inhibited the increase of total cells, immune cells (lymphocyte, neutrophils, macrophage, CD4+, CD8+, CD4+CD69+, CD62L-CD44high+, Gr-1+SiglecF-), and the expression of cytokines (IL-4, IL-5, IL-13, IFN-γ) in BALF. It also significantly inhibited the increase of total cells, immune cells (lymphocyte, neutrophils, eosinophil/macrophage, CD3+, CD19+, CD3+CD193+, CD4+, CD8+, CD4+CD69+, CD62L-CD44high+, and Gr-1+SiglecF-), and the expression of IL-13, TARC, and MCP-1 in lung tissue. GGX decreased the severity of histological lung injury and the expressions of STAT3 and GATA3. Conclusion: This study suggests the probability of using GGX for the treatment of asthma by inhibiting inflammatory immune response.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.4939-4941
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• 2012

Background: The serum level of soluble CD30 (sCD30) is known to be increased with several lymphomas and to correlate with prognosis. Primary effusion lymphoma (PEL) is a highly aggressive malignant lymphoma with poor prognosis, but the existence and significance of sCD30 in PEL have not yet been investigated in detail. Objectives: Since the membrane type of CD30 is frequently expressed on the surface of PEL cells, we compared the expression of the membrane type of CD30 and the production of sCD30 among PEL cell lines as well as other lymphomas. Methods: The expression of surface CD30 in various lymphoma cell lines was analyzed with flow cytometry ans sCD30 was quantified by ELISA. Results: Both surface and sCD30 were detected on PEL cell lines as well as on Hodgkin's lymphoma and adult T-cell leukemia/lymphoma cell lines. Surface CD30 and sCD30 levels of each cell lines correlated with each other. Conclusion: The serum level of sCD30 appear to be a useful biological tumor marker for the diagnosis and management of CD30-positive PEL.

• 대한한의학회지
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• 제29권2호
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• pp.7-20
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• 2008

Objectives: The purpose of this study wasto examine the effects of Kamijihwang-tang (KJHT) extracts on immune cells and cytokines in ovalbumin (OVA)-induced asthmatic mice. Methods: C57BL/6 mice were injected, inhaled and sprayed with OVA for 12 weeks (four times a week) for asthma induction. Two experimental groups were treated with different concentrations of KJHT (400 mg/kg and 200 mg/kg) extracts and cyclosporine A (10 mg/kg) for the later 8 weeks. At the end of the experiment, the mice lung, PLN and spleen were removed and immune cells were analyzed by flow cytometer, IL-5, IL-13, eotaxin-2, $TNF-{\alpha}$ were analyzed by real-time PCR, serum histamine was analyzed by ELISA kit. Results: $CD3^+$, $CD3e^-/CCR3^+$, $CD3e^+/CD69^+$, $CD4^+/CD25^+$, $B220^+/IgE^+$, and $CD3e^+/DX5^+$ cells in lung, PLN, and spleen of the KJHT group (400 mg/kg) decreased compared with that of the control group. $CD3e^+/CD69^+$, $CD4^+/CD25^+$, and $CD3e^+/DX5^+$ cells in lung, PLN, and spleen of the KJHT group (200 mg/kg), CD3+, $CD3e^-/CCR3^+$ cells in lung and PLN of the KJHT group (200 mg/kg) and $B220^+/IgE^+$ cells in lung and spleen of the KJHT group (200 mg/kg) decreased compared with that of the control group. mRNA expression of IL-5, IL-13, eotaxin-2, and $TNF-{\alpha}$ in lung tissue of the KJHT groups (400 mg/kg and 200 mg/kg) decreased compared with that of the control group. Histamine in serum of the KJHT group (400 mg/kg) decreased compared with that of the control group. Conclusions: These results suggest that KJHT can be utilized effectively in treating asthma because it significantly reduces inflammatory cells and cytokines.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제27권1호
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• pp.26-36
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• 2024

Purpose: We investigated the role of CD8⁺T cells as host immune factors in pediatric patients with Helicobacter pylori gastritis. Methods: Gastric mucosal tissue and blood samples were collected from 39 children, including 11 children with H. pylori infection and 28 children as controls. Anti-CD8 and anti-T-bet antibodies were used for immunohistochemistry of the gastric mucosa. For the cell surface and intracellular staining, peripheral blood mononuclear cells were stained with anti-IL7Rα, anti-CX3CR1, anti-CD8, anti-T-bet, and anti-IFN-γ antibodies. Cytokines of sera such as tumor necrosis factor alpha (TNF-α) and CX3CL1 were analyzed using enzyme- linked immunosorbent assay (ELISA). Results: In the immunohistochemistry of gastric mucosa, the frequency of CD8⁺ and T-bet⁺ T cells cells was higher in the H. pylori-positive group than in the control group (26.9± 7.8% vs. 16.9±3.3%, p<0.001; 5.0±2.5% vs. 2.2±0.7%, p=0.001). Between the control and H. pylori-positive groups, the frequency of IL-7Rα^lowCX3CR1⁺ CD8⁺ and T-bet⁺ INF-γ⁺ CD8⁺ T cells were not significantly different between surface and intracellular staining, respectively (40.4±24.0% vs. 38.2±17.8%, p=0.914; 40.4±24.0% vs. 38.2±17.8%, p=0.914). In the ELISA, no significant differences in TNF-α and CX3CL1 concentrations were observed between the control and H. pylori-positive groups (34.3±12.1 pg/mL vs. 47.0±22.6 pg/mL, p=0.114/0.5± 0.1 pg/mL vs. 0.5±0.1 pg/mL, p=0.188). Conclusion: CD8⁺ T and Th1 cells, which secrete IFN-γ, might play important roles in the mucosal immunity of the stomach in children with H. pylori infection.

• 한국식품과학회지
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• 제35권2호
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• pp.291-296
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• 2003

된장 및 간장의 chitooligosaccharide(COS) 함량을 높이기 위해 Bacillus subtilis와 Rhizopus oryzae를 접종하여 koji를 만들고 이들 균주들에 의한 된장 및 간장에서 COS 함량변화에 대하여 알아보았다. COSM에 특이적인 항체를 이용하여 competitive direct ELISA(cdELISA)로 COS를 검출하였는데, 검출 감도는 $0.001{\sim}1{\mu}g/mL$이었고, 회수율은 된장 및 간장에서 각각 102%, 115%를 보였다. B. subtilis koji와 R. oryzae koji를 섞어 담근 된장 또는 간장(BR 된장 또는 간장)이 높은 COS 함량을 보였는데, 두 달 숙성에서 된장이 $171{\mu}g/g$, 간장이$29{\mu}g/mL$을 나타내었다. 일본, 국내 시판, 그리고 실험실에서 만든 된장 및 간장의 COS 함량을 비교해 본 결과, BR 시료구가 일본이나 국내 시판용보다 훨씬 높은 함량을 나타내 주어서 본 실험에서 koji 제조시 이용된 Bacillus subtilis와 Rhizopus oryzae 의해 COS의 함량을 높일 수 있는 가능성을 보여주었다.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.877-880
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• 2004

In this study, we determined the content of chitooligosaccharides (COS) in Korean commercial salt-fermented shrimps by competitive direct enzyme-linked immunosorbent assays (cdELISAs), using anti-COS mixture (COSM) antibody and COSM horseradish peroxidase (HRP) conjugate. When COS6 was spiked into salt-fermented shrimps at the level of $10-300\mu{g/g,}$ the average recovery was $120\pm19%$ ($mean\pmS.D.$). The COS contents of the 92 samples of Korean commercial salt-fermented shrimps collected during February 2000 and August 2002 were $36.3\pm20.7\mug$ COS6 equivalent/g (expressed as "$\mug/g$" hereafter). Among the samples, the COS contents of yuk-jeot ( $40.3 \pm 22.5 \mug/g, n=27$) and buksaewoojeot ($40.2 \pm 21.6 \mug/g, n=5$) were higher than the others. The COS contents of salt-fermented shrimps produced at Gwangcheon ($47.1 \pm 20.7 \mug/g, n=18$) and Gomso ($44.1 \pm 21.8 \mug/g, n=6$) areas were higher than those produced at the other areas. This is the first report to determine COS of salt-fermented shrimps by cdELISA.

• IMMUNE NETWORK
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• 제11권5호
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• pp.268-280
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• 2011

Background: Dengue virus, which belongs to the Flavivirus genus of the Flaviviridae family, causes fatal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) with infection risk of 2.5 billion people worldwide. However, approved vaccines are still not available. Here, we explored the immune responses induced by alternating prime-boost vaccination using DNA vaccine, adenovirus, and vaccinia virus expressing E protein of dengue virus type 2 (DenV2). Methods: Following immunization with DNA vaccine (pDE), adenovirus (rAd-E), and/or vaccinia virus (VV-E) expressing E protein, E protein-specific IgG and its isotypes were determined by conventional ELISA. Intracellular CD154 and cytokine staining was used for enumerating CD4+ T cells specific for E protein. E protein-specific CD8+ T cell responses were evaluated by in vivo CTL killing activity and intracellular IFN-${\gamma}$ staining. Results: Among three constructs, VV-E induced the most potent IgG responses, Th1-type cytokine production by stimulated CD4+ T cells, and the CD8+ T cell response. Furthermore, when the three constructs were used for alternating prime-boost vaccination, the results revealed a different pattern of CD4+ and CD8+ T cell responses. i) Priming with VV-E induced higher E-specific IgG level but it was decreased rapidly. ii) Strong CD8+ T cell responses specific for E protein were induced when VV-E was used for the priming step, and such CD8+ T cell responses were significantly boosted with pDE. iii) Priming with rAd-E induced stronger CD4+ T cell responses which subsequently boosted with pDE to a greater extent than VV-E and rAd-E. Conclusion: These results indicate that priming with live viral vector vaccines could induce different patterns of E protein-specific CD4+ and CD8+ T cell responses which were significantly enhanced by booster vaccination with the DNA vaccine. Therefore, our observation will provide valuable information for the establishment of optimal prime-boost vaccination against DenV.

• 한국식품과학회지
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• 제28권5호
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• pp.953-958
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• 1996

발암성 진균 독소의 하나인 fumonisin을 신속, 간편하게 분석할 수 있는 효소면역측정법 (ELISA)을 개발하고자 하였다. 먼저 Fumonisin $B_1\;(FB_{1})$에 특이적인 항체를 생산하기 위하여 $FB_1$을 keyhole lympet hemocyanin (KLH)에 공유결합시키고 이를 Freund's adjuvant와 함께 토끼에 3차례 피하주사하여 면역하였다. 높은 항체가와 가장 양호한 경합반응을 보인 항혈청으로부터 항체를 정제하였으며, 이 항체의 fumonisin 유사독소에 대한 교차반응은 fumonisin $B_1,\;B_2$및 $B_3$에 대하여 각각 100%, 69% 및 166%이었다. $FB_1$의 검출을 위하여 확립한 직접법 경합ELISA (cdELISA)로 옥수수에 인위적으로 오염시킨 $FB_1$을 정제없이 ELISA로 분석하는 경우, 75% methanol로 시료의 추출 후 완충액으로 1/100 희석하였을 때 양호한 회수율을 보였다. 이 조건하에서 행한 ELISA의 FB₁분석회수율은 $1-30\;{\mu}g/g$의 오염농도범위에서 평균 67%이었으며, 농도별 회수율의 분산은 3.4%로 분석치가 매우 안정하였다. 본 연구에서 개발한 fumonisin 분석용 ELISA system은 수입 및 국내산 농산물로부터 이 독소의 정제없이 손쉽게 다량의 시료를 일차로 분석하는 데 유용할 것으로 판단된다.

• Environmental Analysis Health and Toxicology
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• 제12권3_4호
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• pp.15-22
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• 1997

This experiment was performed to examine the immuno-reactive characteristics of fish metal-binding protein, metallothionein (MT), and gain the practical understandings for the proposed use of fish MT as a biomarker. Liver MT induced by Cd in the silver carp was seperated and purified by gel filtration chromatography and ion exchange chromatography. The immuno-reactivity of fish MT was examined with 3 rabbit antisera. Fish MT showed little reactivity with rabbit anti-rat MT antiserum and a weak reactivity with anti-MT peptide antiserum while showed a strong reactivity with rabbit anti-fish MT antiserum. The time-course change of liver MT in the silver carp, after waterborne exposure to 1 ppm of Cd, was checked by Cd-hem method and established competitive ELISA. In both cases, the induction of liver MT showed a good increasing relationship with the exposure days. The results indicate that the fish MT can be developed as a useful biomonitoring means in the toxicological study and for the evaluation of water pollution.