uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
Characterization of CMVpp65-specific CD8+ T lymphocytes using MHC tetramers in kidney transplant patients and healthy participants
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Oncology, Radiology and Clinical Immunology. (KITM)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences. (Friman, infektion)
Show others and affiliations
2000 (English)In: Transplantation, ISSN 0041-1337, E-ISSN 1534-6080, Vol. 69, no 11, 2243-2250 p.Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Cytomegalovirus (CMV) is a ubiquitous herpesvirus that infects 50-90% of individuals in different populations. After primary infection, the virus persists latently in myeloid cells under the control of specific T-cells. Reactivation of CMV infection may cause lethal organ dysfunction and is frequently seen in immunosuppressed individuals. CD8+ cytotoxic T-cells (CTL) have a primary role in suppressing CMV reactivation, and the dominating CTL response is directed against pp65.

METHODS: MHC tetramers, that is, complexes between HLA class I (or class II) molecules and antigenic peptides conjugated to fluorochromes allow the direct visualization of antigen-specific receptor-carrying T-cells using flow cytometry. We constructed a novel MHC tetramer for identification of CMVpp65-specific CD8+ T-cells using HLA-A2 molecules folded with the immunodominant NLVPMVATV peptide.

RESULTS: The A2/pp65 tetramer specifically stained CMV-directed T-cell lines, and sorted cells showed CMV-specific cytotoxicity. High proportions (0.1-9%) of the CD8+ T-cells were A2/pp65 tetramer+ in healthy HLA-A2+ CMV carriers and in immunosuppressed kidney transplant patients with latent infection. Patients with reactivated CMV infection exhibited up to 15% A2/pp65 tetramer+ cells, which seemed to correlate with CMV load over time. A2/pp65 tetramer+ cells expressed T-cell activation markers.

CONCLUSIONS: The construction of a novel A2/pp65 MHC tetramer enabled the design of a rapid and precise flow cytometric method allowing quantitative and qualitative analysis of CMV-specific T-cells. The number of A2/pp65 tetramer binding CTLs in blood may prove to be clinically relevant in assessing the immune response to CMV.

Place, publisher, year, edition, pages
2000. Vol. 69, no 11, 2243-2250 p.
National Category
Medical and Health Sciences
URN: urn:nbn:se:uu:diva-90462PubMedID: 10868621OAI: oai:DiVA.org:uu-90462DiVA: diva2:162823
Available from: 2003-05-16 Created: 2003-05-16 Last updated: 2010-09-09Bibliographically approved
In thesis
1. Cellular Immune Responses to Allografts and Cytomegalovirus
Open this publication in new window or tab >>Cellular Immune Responses to Allografts and Cytomegalovirus
2003 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Today the immunosuppressive treatment is kept to a level were the incidence of acute rejection is below 20% within the first year after transplantation. As a consequence, a group of transplanted patients is over-immunosuppressed and at risk for infections. There is therefore an urgent need for tools which are able to determine the cellular immune response after organ transplantation. This knowledge would facilitate the task of prospectively opimize the immunosuppressive treatment and identify patients at risk of developing rejection episodes or infections.

To address this issue, a rat-kidney transplantation model for acute rejection was developed to study immune responses to allografts. Infiltrating lymphocytes were analysed using an in vitro culture system which allowed cells to propagate from the biopsies to culture medium. The numbers of outgrowing cells were correlated with morphological and immunohistochemical signs of rejection. When immunosuppressive treatment was administered for 2 and four days after acute rejection, histology did not reveal any improvement, however cellular propagation was reduced by 50 and 75%, respectively. Using the tissue culture technique in human transplanted kidney grafts, which was originally developed for the animal model, the number of propagated cells measured was profoundly higher in grafts with acute cellular rejection than from grafts in other groups. In some cases the number of propagated cells was better correlated with the clinical outcome than the diagnosis made by morphological evaluation. To determine immune responses to cytomegalovirus (CMV), we utilised Human Leukocyte Antigen (HLA) tetramer staining and stimulation of T cells with viral antigens. Both of these techniques independently detected CMV specific T cells in immunosuppressed and healthy individuals with latent or active infection. Although the frequency of CMV specific T cells did not differ between groups, there was a functional impairment in immunosuppressed patients as evidenced by reduced interferon-gamma production. In conclusion, these techniques can be used to determine the cellular immune response to allografts and cytomegalovirus and prove valuable for the optimization of immunosuppressive protocols and antiviral treatment.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2003. 100 p.
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 0282-7476 ; 1269
Immunology, Transplantation, Rejection, Monitoring, Flow Cytometry, MHC, Tetramer, CMV, infection, Immunologi
National Category
Immunology in the medical area
urn:nbn:se:uu:diva-3441 (URN)91-554-5652-9 (ISBN)
Public defence
2003-06-06, Fåhraeussalen, C5, Rudbecklaboratoriet, Uppsala, 13:15
Available from: 2003-05-16 Created: 2003-05-16Bibliographically approved

Open Access in DiVA

No full text


Search in DiVA

By author/editor
Tötterman, Thomas H.
By organisation
Department of Oncology, Radiology and Clinical ImmunologyDepartment of Medical SciencesDepartment of Surgical Sciences
In the same journal
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 242 hits
ReferencesLink to record
Permanent link

Direct link