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Identification of molecular markers for human thymic regulatory CD4+CD25+ T cells by phage display

G L Porto R J Giordano; L C Marti; Carlos Alberto Moreira-Filho; O K Okamoto; V Coelho; Meeting of the Brazilian Society for Immunology (31. 2006 Búzios)

Abstracts São Paulo, SP: Brazilian Society for Immunology, 2006

São Paulo 2006

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  • Título:
    Identification of molecular markers for human thymic regulatory CD4+CD25+ T cells by phage display
  • Autor: G L Porto
  • R J Giordano; L C Marti; Carlos Alberto Moreira-Filho; O K Okamoto; V Coelho; Meeting of the Brazilian Society for Immunology (31. 2006 Búzios)
  • Assuntos: IMUNOLOGIA
  • É parte de: Abstracts São Paulo, SP: Brazilian Society for Immunology, 2006
  • Notas: Disponível em CD-ROM
  • Descrição: Introduction and objectives: In spite of significant advances in the understanding of the biological function of CD4+CD25+ regulatory T cells in the last years, much still remains unknown. One important limitation for the research of these cells is the lack of a specific marker, or a combination of markers, that distinguish them from activated cells and memory T cells. The need for such markers becomes especially apparent when investigating the role of the CD4+CD25+ regulatory T cells in humans. Our aim is to identify novel specific cellular markers for CD4+CD25+ regulatory T cells and to identify thymic molecules that may be potentially relevant to their development, using the phage display technique. Methodology: In our experiments, thymic specimens obtained from children who underwent corrective cardiac surgery at the Heart Institute (InCor), were used to obtain thymic tissue, thymocytes and CD4+CD25+ thymic cells. At the first stage, phage peptide-library was incubated with a pool of PBMC (pre-clearing). After the incubation, phages bound to PBMC were discarded. In the second stage, unbound phages were incubated with either total thymocytes or CD4+CD25+ thymic cells sorted by FACSAria. The phages were recovered in E. coli culture and used in further selections as described by the BRASIL method. After three rounds of selection, the phages recovered were sequenced and their ligands identified. To search for phages that
    presented peptides with specificity for the thymic tissue, peptide-library was incubated with thymic medullar region. The phages were recovered in E. coli culture and used in additional cycles of selection. After three rounds of selection, phages recovered were sequenced and their ligands identified. Results: Until now, we have analyzed the sequences of phages derived from the total thymocytes and phages from thymic tissue. Among them, we identified 2 phages that represent ligands with special interest in immune regulation. Their preferential binding to thymocytes was confirmed by binding assays. We are now analyzing phages obtained after selection with CD4+CD25+ thymocytes.
  • Editor: São Paulo
  • Data de criação/publicação: 2006
  • Formato: res. IR.052.
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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