• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Role of IL-5 and Receptor System in the Regulation of the Immune System and Inflammatory Response.

Research Project

Project/Area Number 02404033
Research Category

Grant-in-Aid for General Scientific Research (A)

Allocation TypeSingle-year Grants
Research Field Immunology
Research InstitutionKumamoto University

Principal Investigator

TAKATSU Kiyoshi  Kumamoto University, Professor, 医学部, 教授 (10107055)

Co-Investigator(Kenkyū-buntansha) 高木 智  熊本大学, 医学部, 特別研究員
HITOSHI Yasumiti  Kumamoto University, Lecturer, 医学部, 助手 (10222241)
TOMINAGA Akira  Kumamoto University, Associate Professor, 医学部, 助教授 (50172193)
YAMAGUCHI Naoto  Kumamoto University, Lecturer (00166620)
ONOUE Kaoru  Kumamoto University, Professor (60037497)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥17,800,000 (Direct Cost: ¥17,800,000)
Fiscal Year 1991: ¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1990: ¥11,200,000 (Direct Cost: ¥11,200,000)
KeywordsInterleukin-5 (IL-5) / B Cell Differentiation / Cytokine / LY-1 B Cells / Growth Factor / Eosinophils / IL-5 Receptor / ab Heterodimer / サイトカイン / サイトカインレセプタ- / クラス変換因子 / 発現クロ-ニング法 / 自己抗体 / 好酸球増多
Research Abstract

(1) We reported the establishment of IL-5 and stromal cell-dependent Ly-l^+ early B cell line in long-term bone marrow culture system (Tominaga et al. Growth Factors 1 : 135, 1989). In this study, we obtained another 8 different cell lines that showed germ line configuration of the J region segments of the IgH genes. Their surface markers were CD45R^+, Ly-l^+, Lyb-2^+, sIgM^-, Thy-l^-, and Mac-l^-. When we treated two of them (J8 and J10) with 5-azacytidine followed by coculture with stromal cells (ST2) and IL-5, they became sIgM^+ B cells and Mac-l^+ macrophage-like cells, respectively. After other early lymphold lines were maintained by coculture with ST2 and IL-5 for more than a year, they showed a heterogeneous DNA rearrangement profile of the J region segment of the IgH gene. Northern blot analysis revealed that these cell lines expressed Cu-mRNA, and 5-mRNA. consistent with normal pre-B cells. Intriguingly, they expressed c-fms-mRNA constitutively. When J13 cells were cocultured … More with ST2 and, GM-CSF ih place of ST2 and IL-5, they acquired Mac-1 expression and retained Ly-1 expression. They were morphologically macrophages, nonspeclfic-esterase positive, and showed phagocytosis of latex beads. The conversion was inhibited by addition of. IL-5. These results support evidence for a close relationship between the myeloid and Ly-1^+ B cell pathways of differentiation, and indicate that our IL-5-dependent clones are multipotential intermediates in differentiation from pre-B cells to B cells and macrophages.
(2) IL-5 has been suggested to be5 lnvolved in in vitro growth and differentiation of B cells and eoginophlls. To envisage the possible engagement of IL-5 in the development of these cells in vivo, transgenic mice carrying the mouse IL-5 gene legated with a metallothionein promoter were generated. The IL-5 transgenic mice exibited elevated levels of IL-5 In the serum and an increase in the levels of serum IgM and IgA. A massive eosinophilia In peripheral blood and spleen and an infiltration of eosinophils In muscle and liver were observed. When cadmium-containing saline was injected i. p. into transgenic mice, a distinctive Ly-l^+ B cell population became apparent in the spleen after 5 days. IL-5 receptors (lL-5R) were detected on those cells by mabs against la-5R. Another interesting finding in these transgenic mice was an increase in polyreactive anti-DNA antibodies of IgM class. It Is suggested, therefore, that aberrant expression of the IL-5 gene may Induce accumulation of Ly-l^+ B cells and eosinophils. Furthermore, this IL-5 transgenic mouse can be a model mouse for eosinophilia and we can determine the role of IL-5 in the differentiation of Ly-l^+ B cells and eosinophils by using this mouse.
(3) We have Isolated CDNA clones encoding a murine IL-5R by expression screening of a library prepared from a murine IL-5-dependent early B cell line. A CDNA library was expressed In COS7 cells and screened by panning with the use of anti-IL-5 receptor monocional antibodies. The deduced amino acid sequence analysis demonstrates that the receptor Is a glycoprotein of 415 amino acids (Mr 45, 284), Including an N-terminal hydrophobic region (17 amino acids), a glycosylated extracellular domain (322 amino acids), a single transmembrane segment (22 amino acids) and a cytoplasmic tall (54 amino acids). COS7 cells transfected with the CDNA expressed a 60-kD protein that bound IL-5 with a single class of affinity (K_D = 2-10 nM). FDC-PL cells transfected with the CDNA for murine IL-5R showed the expression of IL-5 binding sites with both low (K_D=6 nM) and high affinity (K_D=30 pill) and acquired responsiveness to IL-5 for proliferation, although parental FDC-PL cells did not show any detectable IL-5 binding. Northern blot analysis showed that two species of mRNAs (5.0 kb and 5.8 kb) were detected In cell lines that display binding sites for murine IL-5. Homology search for the amino acid sequence of the IL-5 receptor reveals that the IL-5 receptor contains a common motif of a cytokine receptor family that is recently identified.
(4) The murine high-affinity IL-5R consists of at least two membrane polypeptide chains ; a chain of p6O and B chain of P130/Pl4O. We found that the B chain is constitutively expressed in IL-3-dependent early B cell-line. To evaluate whether a component of IL-3R is the B chain of the high affinity IL-5R, effect of anti-IL-3 receptor mAb (anti-Aic-2 mAb) on DNA synthesis of Y16 was examined. Anti-Aic-2 mAb that immunoprecipitated p13O/p14O in the cell lysates of Y16 cells partially inhibited the IL-5- and IL-3-induced proliferation of Y16 cells, whereas it did not react with the a chain of IL-5R. Since anti-Aic-2 mAb is reported to react with. recombinant AIC2A and AIC2B, we examined whether AIC2A or AIC2B is involved in the formation of the high affinity IL-5R. The expression of the murine IL-5R receptor a chain with AIC2B, but not with AIC2A in L cells resulted in the high affinity IL-5 binding sites. The expression of AIC2B alone did not show any IL-5 binding by itself. Cell lysates of these transfectants were crosslinked to form the complex of a and B chains with radiolabeled IL-5. These results clearly indicate that B chain of the murine IL-5 receptor is an indispensable for the formation of high-affinity IL-5 binding sites together with the a chain and that AIC2B is likely the the B chain of the mouse IL-5 receptor. The B chain was found to be identical to the B chain of GM-CSF receptor. Less

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (51 results)

All Other

All Publications (51 results)

  • [Publications] Masahiro Migita,et al.: "Characterization of the human interleukin 5 (IL-5) receptors on eosinophils." Cell.Immunol.133. 484-497 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yasumichi Hitoshi,et al.: "In vivo administration of antibody to murine IL-5 receptor inhibits eosinophilia of IL-5 transgenic mice." Int. Immunol.,. 3. 135-139 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Akira Tominaga,et al.: "Transgenic mice expressing a B cell growth and differentiation factor (interleukin 5) gene develop eosinphilia and autoantibody production." J.Exp.Med.,. 173. 429-438 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Seiji Mita, et al.: "Molecular characterization of the B chain of the murine interleukin 5 (IL-5) receptor." Int. Immunol.3. 665-672 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Satoshi Takaki,et al.: "Identification of the second subunit of the murine interleukin 5 receptor: Interleukin-3 receptor like protein,AIC2B is the component of the high affinity interleukin-5 receptor." EMBO J.10. 2833-2838 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kiyoshi Takatsu.: "Interleukin 5 and its receptor." Microbiol. Immunol.35. 593-606 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Masatoshi Korenaga,et al.: "The role of interleukin-5 (IL-5) in protective immunity to Srtongyloides venezuelensis infection in mice." Immunology,. 72. 502-507 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Naoto Yamaguchi,et al.: "Murine interleukin 5 receptor isolated by immunoaffinity chromatography: comparison of determined N-terminal sequence and deduced primary sequence from cDNA and implication of a role of the intracytoplasmic domain." Int. Immunol.3. 889-898 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yoshiyuki Murata,et al.: "Molecular cloning and characterization of the human interelukin 5 receptor." J.Exp.Med.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Eiichiro Sonoda,et al.: "Differential regulation of IgA production by TGFB and IL-5: TGFB induces class switch of surface IgA^+ B cells, whereas IL-5 promotes postswitch maturation into IgA-secreting cells." Cell. Immunol.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] N. Yamaguchi, et al.: "Characterization of murine IL-5 receptors by using monoclonal anti-receptor antibody." Int. Immunol.2. 181-187 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] A. Tominaga, et al.: "Role of carbohydrates moiety of IL-5 in the expression of biological activity : effect of tunicamhcin on the translation of IL-5 and its Activity." J. Immunol.144. 1345-1352 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y. Hitoshi, et al.: "Distribution of interleukin-5 receptor expression of IL-5 receptor on Ly-1 (CD5)-positive B cells." J. Immunol.144. 4218-4325 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S. Katoh, et al.: "Conversion of Ly-1 B lineage cells to Ly-1 macrophages in long-term bone marrow culture." Develop. Immunol.1. 113-126 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S. Takaki: "Molecular cloning and expression of the murine interleukin 5 receptor." EMBO J.9. 4367-4374 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y. Yamaguchi, et all.: "In vitro and in vivo induction of IL-5 mRNA by IL-2 that causes eosinophiloa in mice." J. Immunol.145. 873-877 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y. Murata, et al.: "Interleukin 5 and interleukin 3 induce serine and tyrosine phosphorylation of several cellular proteins in an interleukin-5 dependent cell line." Bioche. Biophys. Res. Com.173. 1102-1108 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] T. Takahashi, et al.: "Structural comparison of murine T-Cell (B151)-derived IL-5 with rIL-5 : dimer formation is essential for the expression of biological activity." Mol. Immunol.27. 991-920 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H. Enokihara, et al.: "Interleukin 5 activity in sera from patients with eosinophilia." Brit. J. Hematol.75. 458-462 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] M. Migita, et al.: "Characterization of the human interleukin 5 (IL-5) receptors on eosinophils." Cell. Immunol.133. 484-497 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y. Hitoshi, et al.: "In vivo administration of antibody to murine IL-5 receptor inhibits eosinophilia of IL-5 transgenic mice." Int. Immunol.3. 135-139 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] A. Tominaga, et al.: "Transgenic mice expressing a B cell growth abd differentation factor (interleukin 5) gene develop eosinophilia and autoantibody production." J. Exp. Med.173. 429-438 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S. Mita, et al.: "Molecular characterization of the B chain of the murine interleukin 5 (IL-5) receptor." Int. Immunol.3. 665-672 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S. Takaki, et al.: "Identification of the second subunit of the murine Interleukin 5 receptor : Interleukin-3 receptor-like protein, AIC2B is the component of the high affinity interleukin-5receptor." EMBO J.10. 2833-2838 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K. Takatsu: "Interleukin 5 and its receptor." Microbiol. Immunol.35. 593-606 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] M. Korenaga, et al.: "The role of interleukn-5 (IL-5) in protective immunology to Strongyloides Venezuelensis infection in mice." Immunology. 72. 502-507 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] N. Yamaguchi, et al.: "Murine interleukin 5 receptor isolated by immunoaffinity chromatography : comparison of determined N-terminal sequence and deduced primary sequence from cDNA and implication of a role of the intracytoplasmic domain." Int. Immunol.3. 889-898 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y. Murata, et al.: "Molecular cloning and characterization of the human interleukin 5 receptor." J. Exp. Med.175. 341-352 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] E. Sonoda, et al.: "Differential regulation of IgA production by TGFB and IL-5 : TGFB induces class switch of surface IgA+ B cells, whereas IL-5 promotes postswitch maturation into IgA-secreting cells." Cell. Immunol.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Masahiro Migita,et al.: "Characterization of the human interleukin 5 (ILー5) receptors on eosinophils." Cell.Immunol.133. 484-497 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Yasumichi Hitoshi,et al.: "In vivo administration of antibody to murine ILー5 receptor inhibits eosinophilia of ILー5 transgenic mice." Int.Immunol.,. 3. 135-139 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Akira Tominaga,et al.: "Transgenic mice expressing a B cell growth and differentiation factor (interleukin 5) gene deveolp eosinphilia and autoantibody production." J.Exp.Med.,. 173. 429-438 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Seiji Mita,et al.: "Molecular characterization of the B chain of the murine interleukin 5 (ILー5) receptor." Int.Immunol.3. 665-672 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Satoshi Takaki,et al.: "Identification of the second subumit of the murine interleukin 5 receptor: Interleukinー3 receptor like protein,AIC2B is the component of the high affinity interleukinー5 receptor." EMBO J.10. 2833-2838 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kiyoshi Takatsu: "Interleukin 5 and its receptor." Microbiol.Immunol.35. 593-606 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Masatoshi Korenaga,et al: "The role of interleukinー5 (ILー5) in protective immunity to Strongyloides venezuelensis infection in mice." Immunology,. 72. 502-507 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Naoto Yamaguchi,et al.: "Murine interleukin 5 receptor isolated by immunoaffinity choromatography: comparison of determined Nーterminal sequence and deduced primary sequence from cDNA and implication of a role of the intracytoplasmic domain." Int.immunol.3. 889-898 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Yoshiyuki Murata,et al: "Molecular cloning and characterization of the human interelukin 5 receptor." J.Exp.Med.

    • Related Report
      1991 Annual Research Report
  • [Publications] Eiichiro Sonoda,et al.: "Differential regulation of IgA production by TGFB and ILー5: TGFB induces class switch of surface IgA^+ B cells,whereas ILー5 promotes postswitch maturation into IgAーsecreting cells." Cell.Immunol.

    • Related Report
      1991 Annual Research Report
  • [Publications] Naoto Yamagchi,et al.: "Characterization of murine ILー5 receptors by using monoclonal antiーreceptor antibody." Int.Immunol.2. 181-187 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Akira Tominaga,et al.: "Role of carbohydrates moiety of ILー5 in the expression of biological activity:effect of tunicamycin on the translation of ILー5 and its Activity." J.Immunol.144. 1345-1352 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Yasumichi Hitoshi,et al.: "Distribution of interleukinー5 receptorーpositive B cells:expression of ILー5 receptor on Lyー1 (CD5)ーpositive B cells." J.Immunol.144. 4218-4325 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Sigeki Katoh,at al.: "Conversion of Lyー1 B lineage cells to Lyー1 macrophages in longーterm bone marrow culture." Develop.Immunol.1. 113-126 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Satoshi Takaki: "Molecular cloning and expression of the murine interleukin 5 receptor." EMBO J.9. 4367-4374 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Yuji Yamaguchi,et al.: "In vitro and in vivo inductionof ILー5 mRNA by ILー2 that causes eosinophiloa in mice." J.Immunol.145. 873-877 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Yoshiuki MUrata,et al.: "Interleukin 5 and interleukin 3 induce serine and tyrosine phosphorylation of several cellular proteins in an interleukinー5ーdependent cell line." Biochem.Biophys.Res.Com.173. 1102-1108 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Takeo Takahashi,et al.: "Structural comparison of murine TーCell(B151)ーderived ILー5 with rILー5:dimer formation is essential for the expression of biological activity." Mol.Immunol.27. 911-920 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Hideo Enokihara,et al.: "Interleukin 5 activity in sera from patients with eosinophilia." Brit.J.Hematol.75. 458-462 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Masahiro Migita.et al.: "Characterization of the human ILー5 receptors on eosinophils." Cell.Immunol.(1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Yasumichi Hitoshi,et al.: "Ih vivo administration of antibody tomurine ILー5 receptor inhibits eosinophilia of ILー5 transgenic mice." International Immunolol.(1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Akira Tominaga,et al.: "Transgenic mice expressing a B cellgrowth and differentiation factor (ILー5) gene develop eosinophilia and autoantibody production." J.Exp.Med.,. (1991)

    • Related Report
      1990 Annual Research Report

URL: 

Published: 1990-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi