Improved preparation of class I HLA tetramers and their use in detecting CMV-specific CTL.
J Immunol Methods
; 312(1-2): 148-56, 2006 May 30.
Article
em En
| MEDLINE
| ID: mdl-16682054
ABSTRACT
Different methods were used to prepare HLA tetramers and the yields of each method were compared. Our results indicate that preliminary refolding of the heavy chain (Hc) and light chain (beta 2m) yields more monomer than the typical conventional method with urea-solubilized Hc and beta 2m. We then used the corresponding tetramers to detect cytomegalovirus (CMV)-specific cytotoxic T lymphocytes (CTL). Increasing data suggest that the adoptive transfer of CMV-specific CTL constitutes an effective strategy against CMV infections. We designed a method that efficiently induces CMV-specific CTL to a higher frequency in vitro than is currently achieved. This method increased the percentage of CMV-specific CTL from below 1% to 20% of PBL, accounting for more than 40% of CD8+ T cells. Successful HLA tetramer preparation provides the basis for the subsequent detection of CMV-specific CTL in clinical applications.
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Base de dados:
MEDLINE
Assunto principal:
Fosfoproteínas
/
Linfócitos T Citotóxicos
/
Antígenos de Histocompatibilidade Classe I
/
Proteínas da Matriz Viral
/
Infecções por Citomegalovirus
/
Citomegalovirus
/
Antígenos HLA
Limite:
Humans
Idioma:
En
Ano de publicação:
2006
Tipo de documento:
Article