Journal Article

Multimer Staining of Cytomegalovirus Phosphoprotein 65–Specific T Cells for Diagnosis and Therapeutic Purposes: A Comparative Study

Junxia Yao, Clemens Bechter, Markus Wiesneth, Georg Härter, Marlies Götz, Lothar Germeroth, Philippe Guillaume, Ferishte Hasan, Stephanie von Harsdorf, Thomas Mertens, Detlef Michel, Hartmut Döhner, Donald Bunjes, Michael Schmitt and Anita Schmitt

in Clinical Infectious Diseases

Published on behalf of Infectious Diseases Society of America

Volume 46, issue 10, pages e96-e105
Published in print May 2008 | ISSN: 1058-4838
Published online May 2008 | e-ISSN: 1537-6591 | DOI: http://dx.doi.org/10.1086/587749
Multimer Staining of Cytomegalovirus Phosphoprotein 65–Specific T Cells for Diagnosis and Therapeutic Purposes: A Comparative Study

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Background. Cytomegalovirus (CMV) disease represents a serious complication after allogeneic peripheral blood stem cell (PBSC) transplantation. If possible, stem cell donors for transplantation are selected on the basis of their CMV serostatus. However, the cytomegalovirus-specific immune status can be further characterized by measuring CMV phosphoprotein 65–specific CD8+ T cell frequencies using tetramers, pentamers, and streptamers. We therefore investigated the specificity and sensitivity of all 3 methods and compared the results to patient serostatus.

Methods. Twenty-three samples from CMV-seropositive healthy volunteers and 15 samples from CMV-seropositive patients before and after allogeneic PBSC transplantation were stained with tetramers, pentamers, or streptamers and analyzed by flow cytometry.

Results. Similar frequencies of CD8+ and multimer+ T cells could be measured by all 3 multimer technologies. The lowest background signals (⩽0.02%) were obtained using tetramer technology. Frequencies of 0.19%–2.48% of CMV phosphoprotein 65 495–503–specific CD8+ T cells were detected in healthy volunteers. Antigen-specific T cells were detected in only 11 (48%) of 23 seropositive healthy volunteers. CMV antigenemia before day 100 after allogeneic PBSC transplantation occurred in 2 of 3 patients without any specific T cells.

Conclusion. These findings demonstrate the power of multimer staining and a certain limitation of serologic testing to define appropriate donors for transplantation. Therefore, whenever possible, CMV-seropositive donors of transplants to seropositive recipients should be screened for their CD8+ T cell frequency. All 3 multimer technologies can be used, yielding similar results. The streptamer technology additionally offers the advantage of selecting CMV phosphoprotein 65–specific CD8+ T cells at the good manufacturing practice level for adoptive T cell transfer.

Journal Article.  4590 words.  Illustrated.

Subjects: Infectious Diseases ; Immunology ; Public Health and Epidemiology ; Microbiology

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