• Title/Summary/Keyword: Antibodies

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Changing Patterns of Serum and Bile Antibodies in Re-infected Rats with Clonorchis sinensis

  • Zhang, Hong-Man;Chung, Byung-Suk;Li, Shun-Yu;Choi, Min-Ho;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.46 no.1
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    • pp.17-22
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    • 2008
  • Rats develop strong resistance to re-infection and super-infection by Clonorchis sinensis. The present study investigated the antibodies present in the sera and bile juice of rats that were primary infected and re-infected with C. sinensis. The serum level of specific IgG antibodies, which were elevated 2 wk of the primary infection, peaked at 4 wk and subsequently remained unchanged even during re-infection. The total IgE level in serum increased slowly from 388 ng / ml to 3,426 ng / ml beginning 2 wk after the primary infection, and remained high up to 8 wk but dropped to a normal level (259 ng / ml) after treatment. In resistant re-infected rats, the serum IgE level increased rapidly and peaked within 1 wk, whereas no increase was observed in immunosuppressed rats. The serum level of specific IgA antibodies was elevated beginning 1 wk after infection, and decreased 4 wk after treatment. The total bile IgA level unchanged during the primary infection but increased in treated and re-infected rats. The elevated levels of serum IgE and bile IgA indicate that these immunoglobulins may be correlated with the development of resistance to re-infection by C. sinensis in rats.

Analysis of Plant Hormones by Immunoassay I . Production of Monoclonal Antibodies to Indole- 3-acetic acid (면역측정법을 이용한 식물 홀몬의 분석 I. 옥신(IAA)에 대한 단크론 항체 생산)

  • 황태익;임현옥;이재와
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.36 no.6
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    • pp.506-512
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    • 1991
  • Monoclonal antibodies (mAb) to indole-3-acetic acid (IAA) were produced and characterized. Spleen cells from mouse immunized with IAA coupled to bovine serum albumin were fused with SP2/0-Ag14 myeloma cells. Three clones secreted specific antibodies to IAA were established to hybridoma cell lines and designated WLI-G1, WLI-G3 and WLI-Ell. The antibodies produced were classified into IgG, types and revealed the high degree of specificity by cross-reaction in the IAA derivatives and its analogues. In the IAA-ELISA with mAb, the measuring range of the assay was 1-500 p mol, and Ka and binding capacity calculated from Scatchard plot were 6.7 X 10$^{-10}$ L/M and 6 x 10$^{-10}$ L/M respectively. The ELISA with mAb can be used to quantitate IAA directly in crude plant eatract. The results showed that the immunoassay was easy and sensitive method to perform and applicate for quantitative analysis of IAA in plant.

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Production of monoclonal antibodies specific to the surface antigens of chicken peripheral blood mononuclear cells (닭의 혈액내 단핵세포 표면항원 특이 단클론성 항체 생산)

  • Choi, Jun-Gu;Sung, Haan-Woo;Kim, Sun-Joong
    • 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.

Risk Factors for Cholangiocarcinoma in the Lower Part of Northeast Thailand: a Hospital-based Case-control Study

  • Manwong, Mereerat;Songserm, Nopparat;Promthet, Supannee;Matsuo4, Keitaro
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5953-5956
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    • 2013
  • Background: Cholangiocarcinoma (CCA) is the most common cancer in Northeast Thailand. It is also a crucial health problem for Thai people. Various risk factors for CCA have been identified in the upper part of Northeast Thailand, but no similar studies of risk factors have been conducted in the lower parts of the region. This study aimed to investigate factors associated with CCA in the resident population. Materials and Methods: A hospital-based case-control study was conducted during 2009-2012 with the recruitment of 123 CCA cases and 123 non-CCA patient controls, matched for sex, age and residential area. Information was collected by interview with a structured questionnaire. Blood samples were collected for assays of anti-OV antibodies. Associations between various personal factors, dietary habits, family history, the presence of anti-OV antibodies and CCA were analyzed using multiple conditional logistic regression. Results: Patients who consumed raw meat (beef, pork) and alcoholic beverages ${\geq}3$ times per week had a higher risk of CCA than non-consumers ($OR_{adj}$=4.33; 95%CI=1.14-16.35 and $OR_{adj}$=2.13; 95%CI=1.00-4.55, respectively). Patients who had a family history of cancer had a higher risk than those who did not ($OR_{adj}$=4.34; 95%CI=1.80-10.43). Also, patients who had anti-OV antibodies (AU>23.337) had a higher risk than those whose anti-OV antibodies were below the cut-off ($AU{\leq}23.34$) ($OR_{adj}$=3.09; 95%CI=1.04-9.16). Conclusions: As is the case in the upper part of Northeast Thailand, OV infection is a crucial risk factor for CCA in people who live in lower part of the region. Similarly, a family history of cancer and the consumption of alcohol are risk factors for CCA.

Characterization of Carp (Cyprinus carpio L.) Immunoglobulin Structure

  • Choi, Sang-Hoon;Park, Kwan-Ha;Yoon, Jong-Man
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.2
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    • pp.290-296
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    • 2002
  • Serum immunoglobulins (Igs) from Israeli carp were purified using affinity chromatography. Fish were immunized with purified mouse IgG, and the specific fish antibodies were purified from the immune serum on a mouse IgG-immobilized agarose gel. Rabbit anti-Israeli carp Igs (R $\alpha$ I. carp Igs) antibodies were produced following hyperimmunization with mouse IgG specific carp antibodies. SDS-PAGE analysis under reducing condition showed that Israeli carp Igs were composed of two $\mu$-like heavy chains with about 82 and 50 kd, respectively, and one light chain with about 25 kd. On immunoblotting analysis, however, R $\alpha$ I. carp Igs failed to react with the light chain. When both protein A and protein G-purified normal carp Ig were compared with mouse IgG-specific Israeli carp Ig, no significant structural differences among them were observed. To investigate if there is any homology between other fish Ig molecules, cross-reactivity of R $\alpha$ I. carp Igs against Ig molecules from 6 different fish sera and mouse control serum was checked on immunoblotting analysis. As a result, R $\alpha$ I. carp Igs responded to Israeli carp, common carp, and tilapia Ig molecules. In flow cytometry study, however, R $\alpha$ I. carp Igs appeared to recognize 42.0%, 35.8% and <5% of Israeli carp, common carp and tilapia $Ig^+$ head kidney cells, respectively. The result suggests the heterogeneity between receptor Igs on B-like lymphocytes and soluble Igs in serum. It is crucial to obtain pure fish Igs to produce reagent antibodies as tools for the study on their specific immune responses.

Generation of Anti-HLA-DR4 Specific Antibodies by Immunization of the Recombinantly Expressed Allelic Subtype-Specific Region of the $HLA-DRB1^*0405$ Molecules

  • Park, Jung-Hyun;Cho, Eun-Wie;Lee, Yun-Jung;Chung, Jin;Hahm, Kyung-Soo;Kim, Kil-Lyong
    • BMB Reports
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    • v.31 no.2
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    • pp.111-116
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    • 1998
  • HLA-DR4 is the dominant allele of MHC class II genes in Koreans. In particular, the $DRB1^*0405$ subtype has been reported to be almost exclusively expressed in Far East Asians, and has also been observed to be strongly associated with rheumatoid arthritis in Koreans and the Japanese. Identification of this specific allele has been mainly performed by PCR-based methods, which is often time consuming, costly, and involves tedious procedures such as the isolation of genomic DNA, PCR, and gel electrophoresis. To develop a more convenient tool for screening vast amounts of samples as well as to generate reagents which might also be used in other applications, in this study, antibodies were produced against this specific HLA subtype. By PCR, an allelespecific region covering the ${\beta}1$ domain of $DRB1^*0405$ was amplified and recombinantly expressed in E.coli. Immunization of Lewis rats with the purified protein yielded an allele specific antiserum. Western blot analysis showed the selective detection of the HLA-DR ${\beta}-chain$. Using this antiserum, established cell lines and peripheral blood lymphocytes were analyzed on their HLA haplotype by fluorescence activated flow cytometry. These novel antibodies will provide a powerful tool in the detection and investigation of DR4 alleles.

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Studies on In Vitro Fertilization and Development of Bovine Follicular Oocytes Matured In Vitro II. Effect of Anti-Cumulus Cell Antibody on In Vitro Maturation of Bovine Follicular Oocytes (체외성숙 우난포란의 체외수정과 발달에 관한 연구 II. 항란구세포 항체가 우난포란의 체외성숙에 미치는 영향)

  • 박세필;김은영;정형민;박흠대;김종배;정길생
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.93-100
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    • 1990
  • These experiments were carried out to investigate the effect of rabbit anti-bovine cumulus cell antibodies on in vitro maturation of bovine follicular oocytes. Antisera to bovine cumulus cell were produced Japanese Ginat rabbit by repeated immunization of intact or solubilized bovine cumulus cell and purified by ammonium sulfate precipitation and Sepharose CL-4B protein-A affinity chromatography. The bovine cumulus cell-specific antibodies were confirmed by indirect ELISA. The results obtained in these experiments were summarized as follows : 1. The titer of the antibodies to cumulus cell determined by indirect ELISA using intact or solubilized bovine cumulus cell coated plates was very high in both intact and solubilized cumulus cells. Namely, the optical density at 1:12,800 dilution of antibodies was still significantly higher than that of non-immunized control serum. 2. When the follicular oocytes were treated with antibody to intact cumulus cells, the maturation rate of cumulus compacted and removed oocytes was ranged 47.6 to 59.1%. These value is significantly lower(p<0.05) than that(78.8%) of follicular oocytes cultured without the antibody. 3. the maturation rate of cumulus compacted and removed oocytes treated with antibody to solubilized cumulus cells was ranged 46.7 to 59.1%, significantly lower(p<0.05) than that(82.1%) of ooyctes cultured in antibody free medium. From above mentioned results, it could be said that cumulus cells promote nuclear maturation of follicular oocytes and that the beneficial effect of cumulus cells to the oocyte maturation is inhibited by the action of antibody to cumulus cells.

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Development of an Enzyme-Linked Immunosorbent Assay for the Iletection of Aflatoxin $B_1$ (Aflatoxin $B_1$의 검출을 위한 효소면역측정법의 개발)

  • 손동화;박애란;서병철;김진철;이인원;남영중;허우덕
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.225-232
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    • 1992
  • In order to develop an enzyme-linked immunosorbent assay(ELISA) for detecting aflatoxin $B_1(AFB_1)$, we produced and purified antibodies, thereafter established and evaluated methods of direct and indirect competitive ELISA. Anti-AFB, antisera, produced by immunizing rabbits with $AFB_1$-1-(0-carboxymethy1)oxime-bovine serum albumin conjugate ($AFB_1$-BSA), were removed of anti-BSA antibodies by quantitative precipitation reaction and further purified by ammonium sulfate precipitation and DEAE-Sephadex A-50 ion exchange chromatography. Purified IgG fractions were used as anti-$AFB_1$ antibodies. The antibodies, whose titer was deterrnined extremely high above $2 \times 10^6$, showed low cross-reactivity of 3~34% against $AFB_1$ analogues such as G2, B2, and GI. From the standard curves of direct and indirect competitive ELISA for AFBI, the detection ranges were found 0.2~20 and 1~10, 000 ng/ml(ppb) respectively. In their sensitivity, stability, simplicity, and rapidity, the direct method was more suitable than the indirect method for practical use.

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Toxoplasma gondii: Ultrastructural localization of specific antigens and inhibition of intracellular multiplication by monoclonal antibodies

  • Lee, Boo-Young;Ahn, Myoung-Hee;Kim, Hyun-Chul;Min, Duk-Young
    • Parasites, Hosts and Diseases
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    • v.39 no.1
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    • pp.67-76
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    • 2001
  • This experiment was focused on the characterization of anti- Toxoplasma monoclonal antibodies (mAbs) and the effect of mAbs on the parasite invasion of mouse peritoneal macrophages. Twenty eight mAbs including M110, M556, R7A6 and M62l were characterized by Ab titer, immunoglobulin isotyping and western blot pattern. Antibody titer (optical density) of 4 mAbs. Ml 10. M556. R7A6 and M62l. were 0.53,0.67, 0.45 and 0.39 (normal mouse serum; 0.19) with the same IgGl isotypes shown by Enzyme-linked immunosorbent assay (ELISA). Western blot analysis showed that Ml 10. M556. R7A6 and M62l reacted with the 33 kDa (p30),31 kDa (p28),43 kDa and 36 kDa protein. Immuno-gold labelling of mAbs M110, M556, R7A6 and M621 reacted with the surface membrane, dense granules and parasitophorous vacuolar membrane (PVM) , rhoptries and cytoplasm of tachyzoite, respectively. For in vitro assay, preincubation of tachyzoties with four mAbs, Ml 10, M556, R7A6 and M62l resulted in the decrease of the number of infected macrophages (p < 0.05) and the suppression of parasite multiplication at 18 h post-infection. Four monoclonal antibodies including Ml 10 (SAGI) were found to have an important role in the inhibition of macrophage invasion and T. gondii multiplication in vitro, and these mAbs may be suitable for vaccine candidates, diagnostic kit and for chemotherapy.

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Prevalence of antibodies against bovine viral infectious diseases in farmed deer and wild water deer in Jeonbuk province (사육사슴 및 야생고라니의 소 바이러스성 전염병에 대한 혈청학적 연구)

  • Jo, Young-Suk;Chu, Keum-Suk;Lee, Jeong-Won;Camer, Gerry A;Chekarova, Irina;Seol, Min-Suk;Park, Hyun-Jong;Kim, Bum-Seok;Lim, Chae-Woong
    • Korean Journal of Veterinary Service
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    • v.32 no.2
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    • pp.111-117
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    • 2009
  • Farmed deer could be susceptible carrier to bovine viral infectious disease. But unfortunately, there has not been an overall study over this subject in Korea so far. Therefore, a study was conducted to see serum antibodies to bovine leukosis, food and mouth disease, bovine viral diarrhea and infectious bovine rhinotracheitis in deer using the sera of farmed deer. As a result, two deer in a farms showed positive in bovine leukosis antibodies, using ELISA. For wild water deer, no antibodies were found for those diseases. As a result, it can be assumed that deer were relatively low rate of exposure to highly contagious disease such as viral bovine infectious disease in Korea. As this study was conducted over limited in number of subject and regions, continued study should be carried out in order to prevent and control the interspecies transmission in the future.