• Title, Summary, Keyword: Antibody Production

Search Result 726, Processing Time 0.033 seconds

Flagellin-Stimulated Production of Interferon-β Promotes Anti-Flagellin IgG2c and IgA Responses

  • Kang, Wondae;Park, Areum;Huh, Ji-Won;You, Gihoon;Jung, Da-Jung;Song, Manki;Lee, Heung Kyu;Kim, You-Me
    • Molecules and Cells
    • /
    • v.43 no.3
    • /
    • pp.251-263
    • /
    • 2020
  • Flagellin, a major structural protein of the flagellum found in all motile bacteria, activates the TLR5- or NLRC4 inflammasome-dependent signaling pathway to induce innate immune responses. Flagellin can also serve as a specific antigen for the adaptive immune system and stimulate anti-flagellin antibody responses. Failure to recognize commensal-derived flagellin in TLR5-deficient mice leads to the reduction in anti-flagellin IgA antibodies at steady state and causes microbial dysbiosis and mucosal barrier breach by flagellated bacteria to promote chronic intestinal inflammation. Despite the important role of anti-flagellin antibodies in maintaining the intestinal homeostasis, regulatory mechanisms underlying the flagellin-specific antibody responses are not well understood. In this study, we show that flagellin induces interferon-β (IFN-β) production and subsequently activates type I IFN receptor signaling in a TLR5- and MyD88-dependent manner in vitro and in vivo. Internalization of TLR5 from the plasma membrane to the acidic environment of endolysosomes was required for the production of IFN-β, but not for other pro-inflammatory cytokines. In addition, we found that anti-flagellin IgG2c and IgA responses were severely impaired in interferon-alpha receptor 1 (IFNAR1)-deficient mice, suggesting that IFN-β produced by the flagellin stimulation regulates anti-flagellin antibody class switching. Our findings shed a new light on the regulation of flagellin-mediated immune activation and may help find new strategies to promote the intestinal health and develop mucosal vaccines.

CpG-DNA induces bacteria-reactive IgM enhancing phagocytic activity against Staphylococcus aureus infection

  • Kim, Te Ha;Kim, Dongbum;Lee, Heesu;Kwak, Min Hyung;Park, Sangkyu;Lee, Younghee;Kwon, Hyung-Joo
    • BMB Reports
    • /
    • v.52 no.11
    • /
    • pp.635-640
    • /
    • 2019
  • CpG-DNA triggers the proliferation and differentiation of B cells which results in the increased production of antibodies. The presence of bacteria-reactive IgM in normal serum was reported; however, the relevance of CpG-DNA with the production of bacteria-reactive IgM has not been investigated. Here, we proved the function of CpG-DNA for the production of bacteria-reactive IgM. CpG-DNA administration led to increased production of bacteria-reactive IgM both in the peritoneal fluid and serum through TLR9 signaling pathway. When we stimulated B cells with CpG-DNA, production of bacteria-reactive IgM was reproduced in vitro. We established a bacteria-reactive monoclonal IgM antibody using CpG-DNA stimulated-peritoneal B cells. The monoclonal IgM antibody enhanced the phagocytic activity of RAW 264.7 cells against S. aureus MW2 infection. Therefore, we suggest that CpG-DNA enhances the antibacterial activity of the immune system by triggering the production of bacteria-reactive IgM. We also suggest the possible application of the antibodies for the treatment of antibiotics-resistant bacterial infections.

Animal Cell Culture and the Production of Monoclonal Antibody(MAb) Using Biopolymer Membrane (생물고분자 막 형성을 이용한 동물세포 배양 및 단클론항체 생산)

  • 손정화;유선희;김성구
    • KSBB Journal
    • /
    • v.13 no.1
    • /
    • pp.13-19
    • /
    • 1998
  • Biopolymer membrane was prepared using two oppositely charged natural biopolymers. The biopolymer membrane was used for the encapsulation of two hybridoma cell lines(ATCC CRL-1606, ATCC HB-8852) to produce monoclonal antibodies. In order to reduce the down stream steps, the pre size of the membrane was controlled to retain the monoclonal antibodies in the capsules based on the diffusion experiments with standard proteins. T-flask culture showed cell densities of 8$\times$107 cells/mL and 3$\times$107 cells/mL, and MAb concentrations of 506$\mu$g/mL and 109$\mu$g/mL for encapsulated ATCC CRL-1606 and HB-8852, respectively. Two liter perfusion cultures with encapsulated ATCC HB-8852 were performed to enhance the MAb production. The MAb production of the encapsulated hybridoma increased considerably comparing to the culture using silicon tubing for oxygen transfer.

  • PDF

Immune Responses in Broiler Chicks Fed Propolis Extraction Residue-supplemented Diets

  • Eyng, C.;Murakami, A.E.;Santos, T.C.;Silveira, T.G.V.;Pedroso, R.B.;Lourenco, D.A.L.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.28 no.1
    • /
    • pp.135-142
    • /
    • 2015
  • This study was conducted to evaluate the effect of inclusion of propolis extraction residue in the feed of broilers from 1 to 21 d of age on phagocytic activity of macrophages, cutaneous basophil hypersensitivity response to phytohemagglutinin, antibody production against Newcastle disease, lymphoid organ weight and hematological profile and to determine the optimal level of inclusion. 120 chicks, reared in metabolism cages until 21 days of age, were distributed in a completely randomized design, with five treatments (0%, 1%, 2%, 3%, and 4% of propolis residue) and six replications. The relative weight of thymus and monocyte percentage were affected by propolis residue, with a quadratic response (p<0.05) and lowest values estimated at 2.38% and 2.49%, respectively. Changes in relative weight of cloacal bursa and spleen, percentage of lymphocyte, heterophil, basophil, eosinophil, and heterophil:lymphocyte ratio, antibody production against Newcastle disease, phagocytic activity of macrophages and the average number of phagocytosed erythrocytes were not observed. The nitric oxide production with regard to positive control (macrophages+erythrocytes) decreased linearly (p<0.05) with increased doses of propolis residue. The remaining variables of nitric oxide production (negative control - macrophages, and difference between the controls) were not affected by propolis residue. The cutaneous basophil hypersensitivity response to phytohemagglutinin as determined by the increase in interdigital skin thickness exhibited a quadratic response (p<0.05), which predicted a lower reaction response at a dose of 2.60% of propolis residue and highest reaction response after 43.05 hours of phytohemagglutinin injection. The inclusion of 1% to 4% of propolis extraction residue in broiler diets from 1 to 21 days of age was not able to improve the immune parameters, despite the modest changes in the relative weight in thymus, blood monocyte percentage, nitric oxide concentration, and interdigital reaction to phytohemagglutinin.

Studies on Histological Changes of Bursa of Fabricius in Chicken Treated with Thyroxine II. Effect of Thyroxine on Antibody Production (갑상선(甲狀腺) 호르몬이 닭의 Fabricius낭(囊)에 미치는 조직학적변화(組織學的變化)에 관(關)한 연구(硏究) II. 갑상선(甲狀腺) 호르몬이 항체산생(抗體産生)에 미치는 영향(影響))

  • Kim, Soon Bok;Lee, Cha Soo
    • Korean Journal of Veterinary Research
    • /
    • v.20 no.2
    • /
    • pp.99-104
    • /
    • 1980
  • The effects of thyroxine (TX) or propylthiouracil (PPT) administration on the antibody forming activity agains t sheep red blood cell (SRBC) and Newcastle disease virus (NDV) were studied by using of hemagglutination and hemagglutination-inhibition techniques. Antibody titers to both SRBC and NDV increased significantly in the TX-treated group, whereas decreased in the PPT-treated group, compared with control. When TX was administered after antigen inoculatioon, antibody forming activity was significantly enhanced, compared with the TX administration before antigen inoculation.

  • PDF

Effect of Salvia plebeia on IgE antibody mediated allergic reaction in rats

  • Shin, Tae-Yong;Kim, Dae-Keun;Choi, Young-Kyun;Kim, Yong-Jin;Choi, Dong-Sung;Kim, Sang-Hyun;An, Nyeon-Hyung
    • Advances in Traditional Medicine
    • /
    • v.1 no.2
    • /
    • pp.29-35
    • /
    • 2000
  • The effect of aqueous extract of Salvia plebeia R. Br. (Labiatae) (AESP) on immunoglobulin (lgE) antibody mediated allergic reactions in rats was investigated. AESP inhibited passive cutaneous anaphylaxis (PCA) when intravenously, intraperitoneally, and orally administered. AESP dose-dependently inhibited histamine release from rat peritoneal mast cells activated by anti-DNP IgE antibody. Moreover, AESP had an inhibitory effect on anti-DNP IgE antibody induced $TNF-{\alpha}$ production from RPMC. These results suggest that AESP inhibit the IgE-mediated allergic reaction in rats.

  • PDF

Egg Yolk Antibody and Its Application

  • Kim, Mujo;Shinji Higashiguchi;Yoshitomo Iwamoto;Yang, Han-Chul;Cho, hong-Yon;Hsjime Hatta
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.5 no.2
    • /
    • pp.79-83
    • /
    • 2000
  • A han transfers her serum immunoglobulin G to the agg (IgY) and gives immunity to her offspring. Therefore, The hen agg can be an effective supplier of a large amount of antigen specific antibody that accumulates in the egg yolk. Antigen specific antibody has been widely used for immunological analysis in the field of diagnosis as well as pure scientific research. The production and separation technology of IgY is demonstrated in the present study.

  • PDF

Production of Egg Yolk Antibody (IgY) Against Human Placental DNA-Dependent RNA Polymerase II

  • Lee, Yoon-Ik;Surzycki, Stefan S.;Lee, Young-Ik
    • BMB Reports
    • /
    • v.28 no.1
    • /
    • pp.27-32
    • /
    • 1995
  • Polyclonal antibodies against human DNA-dependent RNA polymerase II (HPP II) were generated from chicken egg yolk after immunization with RNA polymerase II as an antigen. The antibodies from egg yolk (IgY) were purified and characterized. IgY showed a specificity against DNA-dependent RNA polymerase II, and was a polyclonal antibody against 12 subunits of polymerase II. An amount of 0.35 mg of IgY was obtained freman HPP II-Sepharose affinity column using 10 eggs from a chicken immunized against RNA polymerase II as an antigen. These antibodies can be used for isolating the genes for RNA polymerase II components, and for in vitro transcription assays using HP-RNA polymerase II.

  • PDF

Purification of Phytase from Aspergillus ficuum and Production of Anti-phytase Antibody (Aspergillus ficuum의 Phytase의 정제와 Anti-phytase 항체생산)

  • Kim, Keun
    • The Korean Journal of Mycology
    • /
    • v.27 no.4
    • /
    • pp.299-303
    • /
    • 1999
  • Phytase(myo-inositol-hexakis phosphate 3-phosphohydrolase, E C 3.1.3.8) sequentially hydrolyzes phytate to myo-inositol and inorganic phosphate. Phytase of Aspergillus ficuum was purified to homogeneity using ultrafiltration, cation exchange column and anion exchange column. It's molecular weight is estimated as around 90,000 by SDS-PAGE. Antibody against the phytase was produced by immunizing mice with the purified phytase. The titer of the antibody was determined to be 1/25,000.

  • PDF

Studies on antigenicity and production of monoclonal antibody for diagnosis of canine heartworm(Dirofilaria immitis) (개 심장사상충(Dirofilaria immitis) 진단을 위한 항원성 조사 및 단크론항체 생산)

  • Lee, Cheol-soon;Jee, Cha-ho
    • Korean Journal of Veterinary Research
    • /
    • v.40 no.1
    • /
    • pp.130-137
    • /
    • 2000
  • In order to diagnose canine heartworm infection by antigen capture ELISA, the crude somatic(S), partial somatic(below 45kDa) and excretory/secretory(E/S) antigen of adult heartworm were identified and the antigenicity was examined by silver stain, immunoblot and ELISA. Then, production of monoclonal antibody to specific antigen carried out in this experiment. The bands to S antigen and E/S antigen were recognized between 10 and 200kDa and common bands were recognized strongly 14, 18, 28, 43kDa by silver stain. By western blot analysis, fractions to S antigen were recognized 14, 16, 18, 20, 24, 28, 32, 43, 50, 55kDa, etc. and only a 14kDa to E/S antigen in positive sera which were positive in modified Knott's test and necropsy. In ELISA, the positive sera reacted to antigens(SA, $SA_{45}$, E/S) were significantly different from negative sera by Student's t-test(p<0.05). Four hybridoma cell lines(14, 16, 17, 32kDa) than produce specific monoclonal antibodies for these antigens were obtained by immunizing BALB/c mice with a partially purified somatic antigen (below 45kDa) preparation, by fusing spleen cells with SP2/O cell myeloma cells, and by screening cell culture supernatants for antibody. In these results, it was confirmed that partial somatic antigen(below 45kDa) or E/S antigen can be used for serologic diagnosis of heartworm infection and monoclonal antibody reacting with specific antigen(14kDa) can be used for antigen capture ELISA in prepatent period of canine heartworm infection.

  • PDF