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- Title
Multimer Staining of Cytomegalovirus Phosphoprotein 65—Specific T Cells for Diagnosis and Therapeutic Purposes: A Comparative Study.
- Authors
Junxia Yao; Bechter, Clemens; Wiesneth, Markus; Härter, Georg; Götz, Marlies; Germeroth, Lothar; Guillaume, Philippe; Hasan, Ferishte; von Harsdorf, Stephanie; Mertens, Thomas; Michel, Detlef; Döhner, Hartmut; Bunjes, Donald; Schmitt, Michael; Schmitt, Anita
- Abstract
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.
- Subjects
LYMPHOCYTES; COMPARATIVE studies; T cells; CELL-mediated lympholysis; PHOSPHOPROTEIN phosphatases; PHOSPHOPROTEINS; PHOSPHATASES; PROTEIN-tyrosine phosphatase; CYTOMEGALOVIRUS diseases
- Publication
Clinical Infectious Diseases, 2008, Vol 46, Issue 10, pe96
- ISSN
1058-4838
- Publication type
Article
- DOI
10.1086/587749