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Real-time imaging analysis of cell adhesion molecules of epidermal keratinocytes

Research Project

Project/Area Number 14370262
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Dermatology
Research InstitutionKeio University

Principal Investigator

NISHIKAWA Takeji  Keio University, School of Medicine, Professor, 医学部, 教授 (50051579)

Co-Investigator(Kenkyū-buntansha) TANAKA Masaru  Keio University, School of Medicine, Associate Professor, 医学部, 助教授 (40188339)
AMAGAI Masayuki  Keio University, School of Medicine, Assistant Professor, 医学部, 専任講師 (90212563)
ISHIKO Akira  Keio University, School of Medicine, Assistant, 医学部, 専任講師 (10202988)
NAGAO Keisuke  Keio University, School of Medicine, Instructor, 医学部, 助手 (40286521)
TAKAE Yujirou  Keio University, School of Medicine, Assistant, 医学部, 助手 (00306367)
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2004: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2003: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2002: ¥7,600,000 (Direct Cost: ¥7,600,000)
KeywordsCell adhesion / Desmoglein / Cadherin / Keratin / Pemphigus / Autoantibody / Adenovirus / Cytoskeleton
Research Abstract

In this study, we developed a novel real-time imaging method with fluorescent-tagged proteins to analyze the functions of desmosomal components, which is critical for cell-cell adhesion between epidermal keratinocytes. Using the technique, we assessed spatial (cellular localization) as well as temporal (variation per hour) movements of the desmosomal components in cultured keratinocytes in response to anti-desmoglein 3 (Dsg3) IgG antibodies in pemphigus. Transformation of GFP-tagged keratin 14 (EGFP-K14) to Normal Human Epidermal Keratinocyte (NHEK) using adenovirus expression system resulted in a reticular network formation of fluorescent keratin fibers in the cytoplasm of NHEK cells. When we shifted the Ca concentration in the culture media to a higher concentration, the fluorescent keratins formed insertion towards cell-cell contact sites. Incubation of anti-Dsg3 IgG antibodies to EGFP-K14-expressed NHEK cells resulted in detachment of keratinocytes as well as in internalization of Dsg3 accompanied by retraction of fluorescent keratin fibers from cell-cell contact sites. These phenomena were determined by either time-lapse imaging analysis or real-time imaging analysis. Furthermore, dynamics of Dsg3 as well as EGFP-K14 in response to anti-Dsg3 IgG were positively influenced by pathogenic strength of the anti-Dsg3 IgG on disruption of cell-cell adhesion of cultured keratinocytes. These findings taken together suggested that our real-time imaging method will give novel insights not only to elucidate the molecular mechanisms for blister formation in pemphigus, but also to cell-cell adhesive functions of epidermal keratinocytes.

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (25 results)

All 2005 2004 2003 Other

All Journal Article (13 results) Publications (12 results)

  • [Journal Article] In vitro keratinocyte dissociation assay for evaluation of the pathogenicity of anti-desmoglein 3 IgG autoantibodies in pemphigus vulgaris.2005

    • Author(s)
      Ishii K, Harada R, Matsuo I, Shirakata Y, Hashimoto K, Amagai M.
    • Journal Title

      J Invest Dermatol 124

      Pages: 939-946

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] In vivo ultrastructural localization of the desmoglein 3 adhesive interface to the ultrastructural localization of the desmoglein 3 adhesive interface to the desmosome mid-line.2005

    • Author(s)
      Shimizu A, Ishiko A, Ota T, Saito H, Oka H, Tsunoda K, Amagai M, Nishikawa T.
    • Journal Title

      J Invest Dermatol 124

      Pages: 984-989

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] In vitro keratinocyte dissociation assay for evaluation of the pathogenicity of anti-desmoglein 3 IgG autoantibodies in pemphigus vulgaris.2005

    • Author(s)
      Ishii K, Harada R, Matsuo I, Shirakata Y, Hashimoto K, Amagai M.
    • Journal Title

      J Invest Dermatol. 124

      Pages: 939-946

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] In vivo ultrastructural localization of the desmoglein 3 adhesive interface to the ultrastructural localization of the desmoglein 3 adhesive interface to the desmosome mid-line.2005

    • Author(s)
      Shimizu A, Ishiko A, Ota T, Saito H, Oka H, Tsunoda K, Amagai M, Nishikawa T.
    • Journal Title

      J Invest Dermatol. 124

      Pages: 984-989

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] In vitro keratinocyte dissociation assay to evaluate pathogenic activity in blister formation by anti-desmoglein 3 IgG autoantibodies in pemphigus vulgaris.2005

    • Author(s)
      Ishii K, Harada R, Matsuo I, Shirakata Y, Hashimoto K, Amagai M
    • Journal Title

      J Invest Dermatol (In press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] In vivo ultrastructural localization of the desmoglein 3 adhesive interface to the desmosome mid-line2005

    • Author(s)
      Shimizu A, Ishiko A, Ota T, Saito H, Oka H, Tsunoda K, Amagai M, Nishikawa T
    • Journal Title

      J Invest Dermatol (In press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] IgG binds to desmoglein 3 in desmosomes and causes a desmosomal split without keratin retraction in a pemphigus mouse model.2004

    • Author(s)
      Shimizu A, Ishiko A, Ota T, Tsunoda K, Amagai M, Nishikawa T.
    • Journal Title

      J Invest Dermatol 122

      Pages: 1145-1153

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Auto-reactive B cells against peripheral antigen, desmoglein 3, escape from tolerance mechanism.2004

    • Author(s)
      Ota T, Aoki-Ota M, Tsunoda K, Simoda K, Nishikawa T, Amagai M, Koyasu S.
    • Journal Title

      Int Immunol 16

      Pages: 1487-1495

    • NAID

      10013567441

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] IgG binds to desmoglein 3 in desmosomes and causes a desmosomal split without keratin retraction in a pemphigus mouse model.2004

    • Author(s)
      Shimizu A, Ishiko A, Ota T, Tsunoda K, Amagai M, Nishikawa T
    • Journal Title

      J Invest Dermatol 122

      Pages: 1145-1153

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Auto-reactive B cells against peripheral antigen, desmoglein 3, escape from tolerance mechanism2004

    • Author(s)
      Ota T, Aoki-Ota M, Tsunoda K, Simoda K, Nishikawa T, Amagai M, Koyasu S
    • Journal Title

      Int Immunol 161

      Pages: 1487-1495

    • NAID

      10013567441

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A mouse model of pemphigus vulgaris by adoptive transfer of naive splenocytes from desmoglein 3 knockout mice2004

    • Author(s)
      Aoki-Ota M, Tsunoda K, Ota T, Iwasaki T, Koyasu S, Amagai M, Nishikawa T
    • Journal Title

      Br J Dermatol 151

      Pages: 346-354

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Conformational epitope mapping of antibodies against desmoglein 3 in experimental murine pemphigus vulgaris2004

    • Author(s)
      Anzai H, Fujii Y, Nishifuji K, Aoki-Ota M, Ota T, Amagai M, Nishikawa T.
    • Journal Title

      J Dermatol Sci 35

      Pages: 133-142

    • NAID

      10016184597

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Induction of pemphigus phenotype by a mouse monoclonal antibody against the amino-terminal adhesive interface of desmoglein 3.2003

    • Author(s)
      Tsunoda K, Ota T, Aoki M, Yamada T, Nagai T, Nakagawa T, Koyasu S, Nishikawa T, Amagai M.
    • Journal Title

      J Immunol 170

      Pages: 2170-2178

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Ota T, Amagai M, Watanabe M, Nishikawa T: "No involvement of IgG autoantibodies against extracellular domains of desmoglein 2 in paraneoplastic pemphigus or inflammatory bowel diseases."J Dermatol Sci. 32. 137-141 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ohyama M, Ota T, Aoki M, Tsunoda K, Harada R, Koyasu S, Nishikawa T, Amagai M.: "Suppression of the immune response against exogenous desmoglein 3 in desmoglein 3 knockout mice : An implication for gene therapy"J Invest Dermatol. 120. 610-615 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Futei Y, Amagai M, Hashimoto T., Nishikawa T: "Conformational epitope mapping and IgG subclass distribution of desmoglein 3 in paraneoplastic pemphigus"J Am Acad Dermatol. 49. 1023-1028 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nishifuji K, Amagai M, Ota T, Nishikawa T, Iwasaki T: "Cloning of canine desmoglein 3 and immunoreactivity of serum antibodies in human and canine pemphigus vulgaris with its extracellular domains."J Dermatol Sci. 32. 181-191 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Shimizu A, Ishiko A, Ota T, Tsunoda K, Amagai M, Nishikawa T: "IgG binds to desmoglein 3 in desmosomes and causes a desmosomal split without keratin retraction in a pemphigus mouse model."J Invest Dermatol. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Aoki-Ota M, Tsunoda K, Ota T, Iwasaki T, Koyasu S, Amagai M, Nishikawa T: "A mouse model of pemphigus vulgaris by adoptive transfer of naive splenocytes from desmoglein 3 knockout mice."Br J Dermatol. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tsunoda K, Ota T, Aoki M, Yamada T, Nagai T, Nakagawa T, Koyasu S, Nishikawa T, Amagai M: "Induction of pemphigus phenotype by a mouse monoclonal antibody against the amino-terminal adhesive interface of desmoglein 3"J Immunol. 170. 2170-2178 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ohyama M, Ota T, Aoki M, Tsunoda K, Harada R, Koyasu S, Nishikawa T, Amagai M: "Suppression of the immune response against exogenous desmoglein 3 in desmoglein 3 knockout mice: An implication for gene therapy"J Invest Dermatol. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kobayashi M, Amagai M, Kuroda-Kinoshita K, Hashimoto T, Shirakata Y, Haslumoto K, Nishikawa T: "BP180 ELISA using bacterial recombinant NC16a protein as a diagnostic and monitoring tool for bullous pemphigoid"J Dermatol Sci. 30. 224-2323 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Cheng SW, Kobayashi M, Kinoshita-Kuroda K, Tanikawa A, Amagai M, Nishikawa T: "Monitoring disease activity in pemphigus with enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3"Br J Dermatol. 147. 261-265 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shimizu A, Ishiko A, Ota T, Tsunoda K, Koyasu S. Amagai M, Nishikawa T: "Ultrastructural changes in mice actively producing antibodies to desmoglein 3 parallel those in patients with pemphigus vulgaris"Arch Dermatol Res. 294. 318-323 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ohyama M, Amagai M, Tsunoda K, Ota T, Koyasu S. Hata J, Umezawa A, Nishikawa T: "Immunologic and histopathologic characterization of an active disease mouse model for pemphigus vulgaris"J Invest Dermatol. 118. 199-204 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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