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2001 Fiscal Year Final Research Report Summary

B cell clone proliferated in HCV infection and V regeon structure of RF correlated to complement clod activation

Research Project

Project/Area Number 11672313
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory medicine
Research InstitutionOsaka Medical College

Principal Investigator

SHIMIZU Akira  Osaka Medical College, Faculty of Medicine Professor, 医学部, 教授 (00028581)

Co-Investigator(Kenkyū-buntansha) NAKANISHI Toyofumi  Osaka Medical College, Faculty of Medicine Assistant Professor, 医学部, 講師 (10247843)
NAKAGAWA Toshimasa  Osaka Medical College, Faculty of Medicine Associate Professor, 医学部, 助教授 (30237226)
Project Period (FY) 1999 – 2001
Keywordscomplement / HCV / RF / immunoglobulin V regeon / mass spectrometry / cold complent activation / cold activation / proteomics
Research Abstract

In vitro activation of the classical pathway of complement at a low temperature is known as cold activation of complete and was first described in the 1970s by Kitamura et al. The former Professor, Shinaya Inai, of the Department of Clinical Pathology, Osaka Medical College, was the leading investigator for this project. We first described the relationship between this phenomenon and HCV infection. Further we found that rheumatoid factor in the serum of hepatitis C virus-infected patients showed an increase in the titer during cold storage. We hypothesized that in the serum of some HVC -infected patients, the affinity of RF appears to be weak, and the binding between the RF and IgG coasted on latex may be inhibited by the complement components of fresh serum.
We tried to analyze the V regeon repertoir of RF obtained from HCV infected and cole activation positive sera. Proteomics strategy was applied to the analysis. Immunogloburin was separated by 2-D electrophoresis on sodium dodecyl sulfate polyacrylamide gels (SDS Page) followed by enzymatic proteolysis of the isolated spot and peptide mapping by mass spectrometry. The germ line sequence with less mutation rate was anticipated. This case the low avidity of RF, which react with ligand (Fc of IgG, aggregated or denatured) only in low temperature.
By these analysis we obtained the sequence, MDMRVPAQLLGLLLLR (Ig k light chain 1-16) ASQSISSYLINWYQQPGKAPK (IG k light chain 47-67)@
DSTYSLSSTLTLSK (Ig k light chain 55-68), These sequence may represent low mutated sequences.
We need more sequence from infected and non-infected which ole activation and without cold activation. Quantitative analysis of protein in ultra mini scale is possible using mass spectrometry and isotope-coded affinity tags. Affinity capture-release electrospray ionization mass spectrometry is also powerful strategy to analyze complex samples in ultra-mini scale. This is also applicable to examine the clonality and mutation diversity.

  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Nakagawa T, Ikemoto, Takeuchi T., Shimizu A: "Eosinophilic peroxidase deficiency : identification of a point mutation (D648N) and prediction of structural changes"Human Mutation. 17・3. 235-236 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Fukada K, Nagano S, Satoh M, Nakanishi T, Shimizu A: "Stability of mutant SOD1 protien by co-expressed wild SOD1 protein causes disease acceleration in familial a amyotrophic lateral sclerosis model mouse"Eur J.Neuroscience. 14・12. 2032-2036 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shimizu A, Nakanishi T, Koyama R, Ikeda T: "Proteomics in Clinical Research : new approach of Mass spectrometry"Rinsyo Byori. 50・2. 169-172 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakanishi T, Koyama R, Ikeda T, Shimizu A: "Catalogue of soluble proteins in the human vitreous humor : comparison between diabetic"J.Chromatography B. (In press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shimizu A, Nakanishi T, Kishikawa M, Miyazaki A: "Detection and identification of protein variants and adducts in blood and tissues : an application of soft ionization mass spectrometry to clinical diagnosis"J.Chromatography B. (In press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakanishi T, Miyazaki A, Shimizu A, Yamaguchi A, Nishimura S: "Assessment of the effect of hemoglobin variants on routine HbA1c measurements by electrospray ionization mass spectrometry"Clinica Chimica Acta. (In press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shimizu A, Nakanishi T, Miyazaki A: "Advances in Mass Spectrometry"Determination of an accurate ratio of a peptide and its adduct by ESIMS : Calculation of the weighted sum of singly and doubly charged ion intensity. 984 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakagawa T., Ikemoto T., Takeuchi T., Shimizu A.: "Eosinophilic peroxidase deficiency: Identification of a point mutation (D648N) and prediction of structural changes"Human Mutation. 17 (3). 235-236 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fukada K., Nagano S., Satoh M., Tohyama C., Nakanishi T., Shimizu A.: "Stability of mutant SOD1 protien by co-expressed wild SOD1 protein causes disease acceleration in familial amyotrophic lateral sclerosis model mouse"Eur. J. Neuroscience. 14 (12). 2032-2036 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shimizu A., Nakanishi T., Koyama R., Ikeda T.: "Proteomics in Clinical Research: new approach of Mass spectrometry"Rinsyo Byori. 50 (2). 169-172 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakanishi T., Koyama R., Ikeda T., Shimizu A.: "Catalogue of soluble proteins in the human vitreous humor: comparison between diabetic retinopathy and macular hole"J. Chromatography B. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shimizu A., Nakanishi T., Kishikawa M., Miyazaki: "A Detection and identification of protein variants and adducts in blood and tissues; an application of soft ionization mass spectrometry to clinical diagnosis"J. Chromatography B. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakanishi T., Miyazaki A., Shimizu A., Yamaguchi A., Nishimura S.: "Assessment of the effect of hemoglobiin variants on routine HbA1c measurements by electrospray ionization mass spectrometry"Clinica Chemica Acta. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shimizu A., Nakanishi T., Miyazaki A.: "Determination of an accurate ratio of a peptide and its adduct by ESIMS: Calculation of the weighted sum of singly and doubly charged ion intensity"Advances in Mass Spectrometry. 984-985 (2001)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2003-09-17  

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