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Wölfl, M.* ; Schalk, S.* ; Hellmich, M.* ; Huster, K.M.* ; Busch, D. ; Berthold, F.*

Quantification of MHC tetramer-positive cells from whole blood: Evaluation of a single-platform, six-parameter flow cytometric method.

Cytometry A 57A, 120-130 (2004)
DOI
Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
Background Quantitation of antigen-specific T cells provides an insight into the development and dynamics of T-cell responses in tumor immunology and infectious diseases. Soluble major histocompatibility class I tetramers are widely used to monitor immune responses; however, variations due to handling and analysis are likely to confound comparisons between different experiments and laboratories. Methods Whole blood from healthy donors was stained with HLA-A*0201/tetramers specific for an epitope of phosphoprotein 65, the immunodominant antigen in cytomegalovirus infection. With the help of Trucount tubes, a single-platform, four-color flow cytometric assay was established to obtain absolute counts of tetramer-positive cells. Various staining and gating strategies were evaluated. Results The no-wash method was a quick and straightforward procedure for the quantitation of tetramer-positive events from whole blood. The level for background staining was low. This information about the intra-assay–related variation and the physiologic variation will allow validation and interpretation of data in future studies. Conclusions The method is highly reliable and can be standardized for multiple experiments. It is therefore suitable for the direct ex vivo analysis of antigen-specific T cells in a variety of clinical settings such as infectious, autoimmune, or neoplastic diseases and can be implemented as a tool for multicenter studies.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Korrespondenzautor
Schlagwörter MHC tetramers; antigen-specific T cells; cytomegalovirus; flow cytometry; single-platform
ISSN (print) / ISBN 1552-4922
e-ISSN 1552-4930
Zeitschrift Cytometry Part A
Quellenangaben Band: 57A, Heft: 2, Seiten: 120-130 Artikelnummer: , Supplement: ,
Verlag Wiley
Verlagsort Hoboken, NJ
Nichtpatentliteratur Publikationen
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Virology (VIRO)