Abu-Khader, Ahmad and Krause, Stefan (2013) Rapid monitoring of immune reconstitution after allogeneic stem cell transplantation - a comparison of different assays for the detection of cytomegalovirus-specific T cells. EUROPEAN JOURNAL OF HAEMATOLOGY, 91 (6). pp. 534-545. ISSN 0902-4441, 1600-0609
Full text not available from this repository. (Request a copy)Abstract
ObjectivesCytotoxic T cells (CTLs) play an important role in controlling cytomegalovirus (CMV) infection after allogeneic hematopoietic stem cell transplantation (HSCT). The purpose of this study was to compare four different assays to detect CMV antigen-specific T cells for the monitoring of CMV-specific immune reconstitution after HSCT. MethodsComparative monitoring of CMV-specific T cells was performed using HLA tetramer, interferon-gamma enzyme-linked immunospot (ELISPOT), intracellular IFN- (IC-FACS), and real-time polymerase chain reaction (RT-PCR) assays with immunodominant CMV pp65-HLA-A2-restricted peptide and recombinant pp65-protein in 18 patients after allogeneic HSCT. ResultsUsing pp65-peptide to stimulate T cells in HLA-A2-positive patients, antigen-specific T cells have been detected in 43% (10 of 23) by IC-FACS, 62% (13 of 21) by tetramer, 68% (15 of 22) by RT-PCR, and 53% (10 of 19) by ELISPOT. No T-cell responses were detected in HLA-A2-negative patients by all assays. Stimulating with combination of both pp65-peptide and pp65-protein detected 72% (18/25) by IC-FACS, 88% (22/25) by RT-PCR, and 59% (12/22) by ELISPOT. Comparing the different methods of T-cell detection we found a significant positive correlation between RT-PCR and tetramer analysis, IC flow cytometry and ELISPOT assay. Using pp65-protein for T-cell activation overcomes the HLA restriction of the HLA-A2-restricted pp65-peptide stimulation, thus increasing the detection limit of antigen-specific T cells. ConclusionCMV-specific T cells have been most frequently detected using RT-PCR suggesting a higher sensitivity compared with IC-FACS, tetramer, and ELISPOT assays. This technique allows rapid and sensitive monitoring of CMV-specific immune reconstitution after HSCT, which could guide prophylactic and therapeutic strategies to avoid active CMV disease.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ENZYME-LINKED IMMUNOSPOT; BLOOD MONONUCLEAR-CELLS; PERIPHERAL-BLOOD; INTERFERON-GAMMA; HEALTHY-INDIVIDUALS; ELISPOT ASSAY; ADOPTIVE TRANSFER; CMV INFECTION; BONE-MARROW; ANTIGEN; allogeneic hematopoietic stem cell transplantation; CMV-specific T cells; monitoring; real-time polymerase chain reaction |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin III (Hämatologie und Internistische Onkologie) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 24 Mar 2020 07:20 |
| Last Modified: | 24 Mar 2020 07:20 |
| URI: | https://pred.uni-regensburg.de/id/eprint/15553 |
Actions (login required)
 |
View Item |
