• Title/Summary/Keyword: IgY (Immunoglobulin Y)

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Kinetic Analysis of CpG-Induced Mouse B Cell Growth and Ig Production

  • Kim, Young-Ha;Lee, Sang-Hoon;Yoo, Yung-Choon;Lee, Jung-Lim;Park, Jong-Hwan;Park, Seok-Rae
    • IMMUNE NETWORK
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    • v.12 no.3
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    • pp.89-95
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    • 2012
  • Immune cells express toll-like receptors (TLRs) and respond to molecular patterns of various pathogens. CpG motif in bacterial DNA activates innate and acquired immune systems through binding to TLR9 of immune cells. Several studies reported that CpG can directly regulate B cell activation, differentiation, and Ig production. However, the role of CpG in B cell growth and Ig production is not fully understood. In this study, we analyzed the effect of CpG on the kinetics of mouse B cell viability, proliferation, and Igs production. Overall, CpG enhanced mouse B cell growth and production of Igs in a dose-dependent manner. Unlike LPS, 100 nM CpG (high dose) did not support TGF-${\beta}1$-induced IgA and IgG2b production. Moreover, 100 nM CpG treatment abrogated either LPS-induced IgM or LPS/TGF-${\beta}1$-induced IgA and IgG2b production, although B cell growth was enhanced by CpG under the same culture conditions. We subsequently found that 10 nM CpG (low dose) is sufficient for B cell growth. Again, 10 nM CpG did not support TGF-${\beta}1$-induced IgA production but, interestingly enough, supported RA-induced IgA production. Further, 10 nM CpG, unlike 100 nM, neither abrogated the LPS/TGF-${\beta}1$- nor the LPS/RA-induced IgA production. Taken together, these results suggest that dose of CpG is critical in B cell growth and Igs production and the optimal dose of CpG cooperates with LPS in B cell activation and differentiation toward Igs production.

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.

A Survey of the Prevalence of Rubella Antibodies in Teachers of Child Bearing Age on Cheju Island (제주도 가임 여교직원의 풍진 항체 보유율 조사)

  • Yang, Hyun-Jong;Hong, Seong-Chul;Bae, Jong-Myon
    • Journal of Preventive Medicine and Public Health
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    • v.33 no.3
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    • pp.280-284
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    • 2000
  • Background : Congenital rubella syndrome (CRS) can be controlled by vaccination. Because rubella is typically a childhood disease, occurring predominantly in the 5 to 14 year age group, female school teachers nay be a high-risk population for CRS. Objectives : To determine the prevalence rate of rubella antibodies in school teachers of child bearing age. Methods : The study population consisted of primary, middle and high school teachers of child bearing age. The subjects were aged 35 years and younger, and consented to immunoglobulin (Ig) level testing using the ELISA method. Results : The positive rate of IgG was 77.9% in the study subjects (n=314). Sixty-three teachers (21.4%) were susceptible to rubella infection. Thirty-seven teachers (11.8%) had a history of rubella vaccination. Among the female teachers with no vaccination history, the proportion of negative IgM and IgG was 21.7%, and the proportion of positive IgM was 2.9%. Seventy-nine percent of the study subjects did not know that they should not become pregnant for three months after receiving the rubella vaccine. Conclusion : School teachers of child bearing age should be considered a high risk group for CRS, and should be vaccinated if they are found to be seronegative.

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Enhanced Delivery of siRNA Complexes by Sonoporation in Transgenic Rice Cell Suspension Cultures

  • Cheon, Su-Hwan;Lee, Kyoung-Hoon;Kwon, Jun-Young;Choi, Sung-Hun;Song, Mi-Na;Kim, Dong-II
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.781-786
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    • 2009
  • Small interfering synthetic double-stranded RNA (siRNA) was applied to suppress the expression of the human cytotoxic-T-Iymphocyte antigen 4-immunoglobulin (hCTLA4Ig) gene transformed in transgenic rice cell cultures. The sequence of the 21-nucleotide siRNA was deliberately designed and synthesized with overhangs to inactivate the expression of hCTLA4Ig. The chemically synthesized siRNA duplex was combined with polyethyleneimine (PEl) at a mass ratio of 1:10 (0.33 ${\mu}g$ siRNA:3.3 ${\mu}g$ PEl) to produce complexes. The siRNA complexes (siRNA+PEI) were labeled with Cy3 in order to subsequently confirm the delivery by fluorescent microscopy. In addition, the cells were treated with sonoporation at 40 kHz and 419W for 90 s to improve the delivery. The siRNA complexes alone inhibited the expression of hCTLA4Ig to 45% compared with control. The siRNA complexes delivered with sonoporation downregulated the production of hCTLA4Ig to 73%. Therefore, we concluded that the delivery of siRNA complexes into plant cells could be enhanced successfully by sonoporation.

The Effects of Oldenlandiae Diffusae Herba Extract on Eosinophil, IgE and IL-4 in Experimental Asthma induced by Ovalbumin (백화사설초(白花蛇舌草)가 ovalbumin으로 유도된 천식동물모델에서 Eosinophil의 수, IgE 및 IL-4에 미치는 영향)

  • Kim, Sang-Chan;Byun, Sung-Hui
    • The Korea Journal of Herbology
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    • v.20 no.2
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    • pp.35-42
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    • 2005
  • Objective : This study was performed to investigate the effect of oral administration of Oldenlandiae Diffusae Herba (ODH) on eosinophil, immunoglobulin-E and interleukin-4 in the experimental asthma induced by ovalbumin. Methods : Asthma was induced to Balb/c mouse by i.p. injection and aerosol immunization with ovalbumin. It was observed the change of the eosinophil number in the BALF. And, it was observed the change of the concentrations of IL-4 in BALF and splenocyte, IgE in serum by ELISA. Results : 1. The number of eosinophil in BALF was significantly decreased in ODH group copared with control group. 2. Concentration of IL-4 in serum, BALF and splenocyte was significantly decreased in ODH group compared with control group, respectively. 3. Level of IgE in serum was significantly decreased in ODH group compared with control group. Conclusion : We found that the effect of ODH extract in asthma was implicated in reductions of IL-4 released from Th2 cell, and decreases of IgE from plasma cell. These findings suggest that ODH extract can produce anti-asthmatic effect, which may playa role in allergen-induced asthma therapy.

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Production and characterization of monoclonal antibodies (MAb) against flounder serum immunoglobulin (Ig)

  • Jang, Han-Na;Cho, Young-Hye;Park, Sang-Hoon
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.446-446
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    • 2000
  • Specific polyclonal and/or monoclonal antibodies (MAbs) to immunoglobulins (Igs) and their subunits have proved to be valuable tools in immunological research and in immunological assays. In this study, we developed and characterized MAbs against flounder serum Igs. To obtain the pure flounder serum Igs, mouse IgG (mIgG) was immunized to flounder. Flounder Igs were purified by using mIgG-agarose affinity column chromatography. The structure of purified flounder Ig was observed, on denatured SDS-PAGE, to be composed of two heavy chains (77 and 72 kd) and two light chains (28 and 26 kd). MAbs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously primed with the flounder Igs. Finally, three hybridoma clones, FIM 511, FIM 519 and FIM 562 were established to recognize both 2 heavy chains, 26 kd of light chain and 28 kd of light chain, respectively. On the other hand, the flounder immune sera collected on the weekly basis were tested on ELISA and immunoblot analysis whether boosting effect is present in flounder humoral immune system. As a result, the secondary immune response in flounder was ascertained on ELISA, but not on immunoblot analysis. Further, we observed an alteration of serum protein levels following immunization. Our MAbs and basic information on flounder humoral immune system obtained in this study will be helpful to control and monitor the efficiency of fish vaccines and therapeutic process of flounder diseases.

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Scutellaria baicalensis Modulates Cytokine Production, T Cell Population and Immunoglobulin Level by Mesenteric Lymph Node Lymphocytes in Experimental Mice with Colitis (한약재인 황금의 궤양성 실험동물에 대한 장간막 임파절 임파구의 면역글로블린 수준, T세포집단, 사이토카인 생성의 조절작용)

  • Lim, Beong-Ou;Park, Pyo-Jam;Choi, Wahn-Soo;Kim, Jong-Dai
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.2
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    • pp.100-105
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    • 2008
  • We previously examined extracts, isolated from Scutellaria baicalensis (SB), chemical mediators, and IgE by mesenteric lymph node (MLN) lymphocytes in rats. The present study was to evaluate the effects of extracts of SB on the MLN lymphocytes function of mice given orally by 20 mg/kg for 2 weeks with dextran sulfate sodium (DS)-induced colitis. Results show that IgE levels in MLN lymphocytes was low, while IgA was high, in mice given SB compared to that fed water. Concentrations of $Inteferon-{\gamma}$ and interleukin (IL)-2 of T cells by concanavalin A treatment was significantly higher in the SB fed group than the normal group. Activation-induced IL-4 and IL-10 secretion was lower in SB fed mice compared control mice after DS-induced colitis. These results suggested that SB suppresses the inflammation in DS-induced colitis through the modulation of Th1/Th2 balance to down-regulate $Th_2$ response in MLN lymphocytes.

Chronologic chrnge of serum IgG antibody response in chickens reinfected with Cryptosporidium baileyi (닭와포자충 재감염닭의 혈청1gG 항체가 추이)

  • Lee, Jae-Gu;Kim, Hyeon-Cheol;Park, Bae-Geun
    • Parasites, Hosts and Diseases
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    • v.34 no.4
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    • pp.255-258
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    • 1996
  • Eight 2-day-old SPF Chickens were each inoculated Orally With 3 Sing1e dose Of 5 × 105 oocysts of Cryptosporinium boilevi. and immunoglobulin G (IgG) antibody responses were chronologically measured by indirect immunofluorescent antibody (IFA) assay. Anti-C. bcileyi IgG antibody levels remained high (1 : 106.67 to 1:512.00) for at least 4 months with 330 days of a detectable period. Ten days after the negative conversion, each chicken was re-challenged with 1 × 107 oocysts of the same species. Subsequent infection in 340-day-old individuals caused sudden elevated IgG antibody levels and the titer peaked on day 28 postchallenge inoculation (PCI), at 1:1.024 with a 65 days of detection period. Chickens in primary infection showed oocyst shedding profiles. but did not exhibit any oocyst shedding before or after experimental reinfection.

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Isolation of Mouse Ig Heavy and Light Chain Genomic DNA Clones, and Construction of Gene Knockout Vector for the Generation of Humanized Xenomouse (인간 단클론 항체 생산용 Humanized Xenomouse 제작의 기초 소재인 생쥐 Ig 중사슬 및 경사슬 Genomic DNA 클론의 확보 및 유전자 적중 벡터의 제작)

  • Lee, Hee-kyung;Cha, Sang-hoon
    • IMMUNE NETWORK
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    • v.2 no.4
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    • pp.233-241
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    • 2002
  • Background: Monoclonal antibodies (mAb) of rodent origin are produced with ease by hybridoma fusion technique, and have been successfully used as therapeutic reagents for humans after humanization by genetic engineering. However, utilization of these antibodies for therapeutic purpose has been limited by the fact that they act as immunogens in human body causing undesired side effects. So far, there have been several attempts to produce human mAbs for effective in vivo diagnostic or therapeutic reagents including the use of humanized xenomouse that is generated by mating knockout mice which lost Ig heavy and light chain genes by homologous recombination and transgenic mice having both human Ig heavy and light gene loci in their genome. Methods: Genomic DNA fragments of mouse Ig heavy and light chain were obtained from a mouse brain ${\lambda}$ genomic library by PCR screening and cloned into a targeting vector with ultimate goal of generating Ig knockout mouse. Results: Through PCR screening of the genomic library, three heavy chain and three light chain Ig gene fragments were identified, and restriction map of one of the heavy chain gene fragments was determined. Then heavy chain Ig gene fragments were subcloned into a targeting vector. The resulting construct was introduced into embryonic stem cells. Antibiotic selection of transfected cells is under the progress. Conclusion: Generation of xenomouse is particularly important in medical biotechnology. However, this goal is not easily achieved due to the technical difficulties as well as huge financial expenses. Although we are in the early stage of a long-term project, our results, at least, partially contribute the successful generation of humanized xenomouse in Korea.

Effects of Excess Dietary Supplementation of Several Micronutrients on Immune Response in Layers Inoculated with Newcastle Disease and Infectious Bronchitis Vaccines (미량영양소들의 추가급여가 뉴캣슬 또는 전염성 기관지염 백신 접종시 산란계의 면역성에 미치는 효과)

  • 김정우;고승연;조석현;김춘수
    • Korean Journal of Poultry Science
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    • v.22 no.2
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    • pp.85-95
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    • 1995
  • This study was conducted to investigate the immune response of layers fed diets supplemented with excess micronutrients, i.e., vitamin A, methionine, Zn, Cu, and Fe to the inoculation of Newcastle disease vaccine(NDV) or infectious bronchitis vaccine(IBV). The antibody titer against the NDV increased immediately after the inoculation and stayed high during the next 6 wk. On the other hand, The antibody titer against the IBV increased after 4 wk of inoculation The IgM level increased rapidly after 1 wk of NDV inoculation, however, it decreased after 5 wk of inoculation. The IgA displayed similar pattern to that of IgM in response to NDV inoculation. The pattern of IgM change after IBV inoculation was similar to that when layers were treated with NDV. However, IgA level changed earlier than did IgM. The IgG response to the NDV and IBV was very weak compared to the other immune responses. The excess supplementation of micronutrients to the diets of layers inoculated with NDV elicited favorable antibody titer and immune response compared to the layers fed the control diet. The excess Zn, however, allowed the layers to have higher antibody titer for the 4-wk period after NDV injection: after that they showed no effect of extra-Zn. The immune responses of layers fed excess vitamin A, Cu, methionine, and Fe were markedly higher in IgA and IgG than the control layers. The excess Zn, however, did not bring about any favorable result. No difference was detected in IgG level between control and micronutrients-treated groups.

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