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Article: A method to increase tetramer staining efficiency of CD8+ T cells with MHC-peptide complexes: therapeutic applications in monitoring cytotoxic T lymphocyte activity during hepatitis B and C treatment

TitleA method to increase tetramer staining efficiency of CD8+ T cells with MHC-peptide complexes: therapeutic applications in monitoring cytotoxic T lymphocyte activity during hepatitis B and C treatment
Authors
Issue Date2004
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jim
Citation
Journal of Immunological Methods, 2004, v. 285 n. 1, p. 71-87 How to Cite?
AbstractThe development of peptide–MHC tetrameric complexes heralds a new era in the study of antigen-specific T cells and their role in viral infections. However, the frequencies of tetramer-staining CD8+ T cells in fresh peripheral blood mononuclear cells (PBMCs) are usually below 1% in patients with chronic hepatitis B and C viruses (HBV and HCV) as well as human immunodeficiency virus (HIV) infections, which makes difficult the comparison and sequential evaluation of different individuals. Thus, the development of a method to enumerate efficiently antigen–specific CD8+ T cells will be clinically beneficial in monitoring the antiviral cellular immunity during therapy. We report here a modified CRI-p culture method (cytotoxic T lymphocyte response index of the epitope–peptide method), using a panel of peptides to stimulate PBMCs in bulk culture. The modified CRI-p cultured cells were, in turn, subjected to fluorescence-activated cell sorter (FACS) analysis, tetramer staining or T-cell functional assays to quantify the antiviral immunity of HLA-A2 (+) HBV and HCV patients receiving antiviral therapies. The results obtained showed that patients with a sustained response had a significantly higher increase in the frequencies of tetramer staining of virus-specific CD8+ T cells than did nonresponders. This method permits semi-quantitative determination of the relative strength of CTL activity against a panel of peptides and provides a large number of cells for FACS analysis from a single blood sampling. Significantly, it achieves high frequencies of tetramer staining of CD8+ T cells allowing the data of different individuals to be easily compared and sequentially evaluated. The mechanisms involved in this method are discussed. Keywords
Persistent Identifierhttp://hdl.handle.net/10722/225164
ISSN
2015 Impact Factor: 1.858
2015 SCImago Journal Rankings: 1.053

 

DC FieldValueLanguage
dc.contributor.authorTsai, SL-
dc.contributor.authorLee, TH-
dc.contributor.authorChien, RN-
dc.contributor.authorLiao, SK-
dc.contributor.authorLin, CL-
dc.contributor.authorKuo, GC-
dc.contributor.authorLiaw, YF-
dc.date.accessioned2016-04-26T04:44:44Z-
dc.date.available2016-04-26T04:44:44Z-
dc.date.issued2004-
dc.identifier.citationJournal of Immunological Methods, 2004, v. 285 n. 1, p. 71-87-
dc.identifier.issn0022-1759-
dc.identifier.urihttp://hdl.handle.net/10722/225164-
dc.description.abstractThe development of peptide–MHC tetrameric complexes heralds a new era in the study of antigen-specific T cells and their role in viral infections. However, the frequencies of tetramer-staining CD8+ T cells in fresh peripheral blood mononuclear cells (PBMCs) are usually below 1% in patients with chronic hepatitis B and C viruses (HBV and HCV) as well as human immunodeficiency virus (HIV) infections, which makes difficult the comparison and sequential evaluation of different individuals. Thus, the development of a method to enumerate efficiently antigen–specific CD8+ T cells will be clinically beneficial in monitoring the antiviral cellular immunity during therapy. We report here a modified CRI-p culture method (cytotoxic T lymphocyte response index of the epitope–peptide method), using a panel of peptides to stimulate PBMCs in bulk culture. The modified CRI-p cultured cells were, in turn, subjected to fluorescence-activated cell sorter (FACS) analysis, tetramer staining or T-cell functional assays to quantify the antiviral immunity of HLA-A2 (+) HBV and HCV patients receiving antiviral therapies. The results obtained showed that patients with a sustained response had a significantly higher increase in the frequencies of tetramer staining of virus-specific CD8+ T cells than did nonresponders. This method permits semi-quantitative determination of the relative strength of CTL activity against a panel of peptides and provides a large number of cells for FACS analysis from a single blood sampling. Significantly, it achieves high frequencies of tetramer staining of CD8+ T cells allowing the data of different individuals to be easily compared and sequentially evaluated. The mechanisms involved in this method are discussed. Keywords-
dc.languageeng-
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jim-
dc.relation.ispartofJournal of Immunological Methods-
dc.subject.meshCD8-Positive T-Lymphocytes - immunology-
dc.subject.meshCytotoxicity Tests, Immunologic - methods-
dc.subject.meshHepatitis B, Chronic - drug therapy - immunology-
dc.subject.meshHepatitis C, Chronic - drug therapy - immunology-
dc.subject.meshT-Lymphocytes, Cytotoxic - immunology-
dc.titleA method to increase tetramer staining efficiency of CD8+ T cells with MHC-peptide complexes: therapeutic applications in monitoring cytotoxic T lymphocyte activity during hepatitis B and C treatment-
dc.typeArticle-
dc.identifier.emailLin, CL: clin@hkucc.hku.hk-
dc.identifier.doi10.1016/j.jim.2003.11.005-
dc.identifier.pmid14871536-
dc.identifier.scopuseid_2-s2.0-1042292703-
dc.identifier.hkuros87731-
dc.identifier.volume285-
dc.identifier.issue1-
dc.identifier.spage71-
dc.identifier.epage87-
dc.publisher.placeNetherlands-

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