Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2003 Apr;132(1):96-104.
doi: 10.1046/j.1365-2249.2003.02098.x.

Cellular responses to cytomegalovirus in immunosuppressed patients: circulating CD8+ T cells recognizing CMVpp65 are present but display functional impairment

M Engstrand  1 , A K Lidehall, T H Totterman, B Herrman, B-M Eriksson, O Korsgren
Affiliations

Cellular responses to cytomegalovirus in immunosuppressed patients: circulating CD8+ T cells recognizing CMVpp65 are present but display functional impairment

M Engstrand et al. Clin Exp Immunol. 2003 Apr.

Abstract

The availability of tetrameric complexes of HLA class I molecules folded with immunodominant peptides makes it possible to utilize flow cytometry for rapid and highly specific visualization of virus specific CD8+ T cells. An alternate technique is to incubate whole blood with specific antigens and to subsequently detect and characterize responding T cells (e.g. by performing intracellular staining of interferon-gamma). By using an HLA-A2 tetramer construct folded with the same immunodominant CMV-peptide as that used for peptide pulsing, we monitored both the presence and functional capacity of CMV-specific CD8+ T cells. In addition T cell activation was assayed by determination of CD38 and CD69 expression. Twelve organ transplant patients and 31 healthy blood donors with latent CMV infection were investigated using CMV pp65 tetramer staining and intracellular staining of interferon-gamma after CMV pp65 peptide pulsing or CMV lysate pulsing. CMV-specific T cells were detected in similar absolute numbers as well as frequencies of T cells in the two groups investigated. However, the CMV-specific CD8+ T cells in immunosuppressed individuals showed a decreased functional response to the CMV-peptide, as evidenced by reduced interferon-gamma production when compared to healthy blood donors (19%; 42%, P < 0.005). In addition, CD38 expression was markedly higher in immunosuppressed patients compared to healthy blood donors (24%; 6%, P < 0.005). In a case report we demonstrate that reactivation of CMV can occur in an immunosuppressed patient with high number of CMV-specific T cells, but without functional capacity. Hence, these findings reflect impaired activation of cytotoxic T cells controlling latent CMV infection in immunosuppressed patients.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Detection of CMV-specific T lymphocytes using flow-cytometry in a healthy HLA A2-positive CMV-seropositive blood donor. T lymphocytes were defined as CD3 high; side-scatter low (a). CMVpp65 tetramer-positive T lymphocytes were all CD8+ (b). IFN-γ-positive T lymphocytes after pulsing with CMVpp65-peptide showing decreased expression of CD8 (c). IFN-γ positive T lymphocytes after pulsing with CMV lysate showing decreased expression of CD4 (d). Unpulsed CD8+ (e) or CD4+ (f) T lymphocytes displaying low background-staining.
Fig. 2
Fig. 2
Comparison of HLA-A2/pp65 tetramer staining with CMVpp65 peptide- and CMV lysate-pulsing techniques for the detection of CMV-specific T lymphocytes in HLA-A2-positive, CMV-seropositive healthy individuals. The frequency of CMV-specific T lymphocytes was highest using the lysate pulsing technique, followed by tetramer staining and CMVpp65 peptide pulsing. Four individuals did not display CMV-specific T cells using any of the techniques.
Fig. 3
Fig. 3
Visualization of a reduced functional capacity of CD8+ CMV-specific T cells in immunosuppressed patients. In HLA-A2-positive and CMV-seropositive individuals, the fraction of tetramer-positive T cells responding to peptide pulsing by IFN-γ synthesis was lower in immunosuppressed patients compared with non-immunosuppressed blood donors. Results are shown as the percentage of CMV pp65 tetramer binding cells and CMV pp65 peptide responding cells for each individual (a). The ratio of CMV tetramer-positive cells responding with IFN-γ secretion was calculated after determining the absolute number of tetramer- and IFN-γ positive cells (b). IS: immunosuppressed (n = 12); non-IS: non-immunosuppressed (n = 12) •, Tetramer staining; ○, peptide pulsing; ▵, ratio.
Fig. 4
Fig. 4
IFN-γ-secreting CD4+ T lymphocytes after pulsing with CMV lysate. In HLA-A2-positive and CMV-seropositive individuals, no significant difference could be seen between immunosuppressed patients compared with non-immunosuppressed blood donors. IS: immunosuppressed (n = 12); non-IS: non-immunosuppressed (n = 12)
Fig. 5
Fig. 5
Expression of CD38 on CMV lysate-responding T lymphocytes. The CD38 expression was markedly higher in immunosuppressed patients than in healthy blood donors (P < 0·005). ▪, Non-immunosuppressed (n = 12); □, immunosuppressed (n = 12).
Fig. 6
Fig. 6
Reactivated CMV infection in CMV-seropositive recipient of a kidney transplant from a CMV-seropositive donor. After CMV reactivation, the absolute number of tetramer-positive cells increases and remain high during antiviral treatment. However, functional response in terms of IFN-γ secretion after peptide or lysate pulsing has not been seen in this patient, suggesting that this individual may have an impaired ability to keep the CMV infection in a latent stage. (a) •, Creatinine (µmol/l); ○, WBC (×109). (b) □, Prednisolone (mg/24 h); •, cyclosporine (ng/ml). (c) ○, CMV–DNA (×103 copies/ml; •, CMVpp65+ (×103/ml); ▪, ganciclovir; □, valaciclovir.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Reusser P, Cathomas G, Attenhofer R, Tamm M, Thiel G. Cytomegalovirus (CMV)-specific T cell immunity after renal transplantation mediates protection from CMV disease by limiting the systemic virus load. J Infect Dis. 1999;180:247–53. - PubMed
    1. Lee PP, Yee C, Savage PA, et al. Characterization of circulating T cells specific for tumor-associated antigens in melanoma patients. Nat Med. 1999;5:677–85. - PubMed
    1. Komanduri KV, Viswanathan MN, Wieder ED, et al. Restoration of cytomegalovirus-specific CD4+ T-lymphocyte responses after ganciclovir and highly active antiretroviral therapy in individuals infected with HIV-1. Nat Med. 1998;4:953–6. - PubMed
    1. Li CR, Greenberg PD, Gilbert MJ, Goodrich JM, Riddell SR. Recovery of HLA-restricted cytomegalovirus (CMV)-specific T cell responses after allogeneic bone marrow transplant: correlation with CMV disease and effect of ganciclovir prophylaxis. Blood. 1994;83:1971–9. - PubMed
    1. Engstrand M, Tournay C, Peyrat MA, et al. Characterization of CMVpp65-specific CD8+ T lymphocytes using MHC tetramers in kidney transplant patients and healthy participants. Transplantation. 2000;69:2243–50. - PubMed

Publication types

MeSH terms