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
. 2005 Sep;79(18):12112-6.
doi: 10.1128/JVI.79.18.12112-12116.2005.

Increased expression of the NK cell receptor KLRG1 by virus-specific CD8 T cells during persistent antigen stimulation

Robert Thimme  1 , Victor Appay, Marie Koschella, Elisabeth Panther, Evelyn Roth, Andrew D Hislop, Alan B Rickinson, Sarah L Rowland-Jones, Hubert E Blum, Hanspeter Pircher
Affiliations

Increased expression of the NK cell receptor KLRG1 by virus-specific CD8 T cells during persistent antigen stimulation

Robert Thimme et al. J Virol. 2005 Sep.

Abstract

The killer cell lectin-like receptor G1 (KLRG1) is a natural killer cell receptor expressed by T cells that exhibit impaired proliferative capacity. Here, we determined the KLRG1 expression by virus-specific T cells. We found that repetitive and persistent antigen stimulation leads to an increase in KLRG1 expression of virus-specific CD8+ T cells in mice and that virus-specific CD8+ T cells are mostly KLRG1+ in chronic human viral infections (human immunodeficiency virus, cytomegalovirus, and Epstein-Barr virus) but not in resolved infection (influenza virus). Thus, by using KLRG1 as a T-cell marker, our results suggest that the differentiation status and function of virus-specific CD8+ T cells are directly influenced by persistent antigen stimulation.

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
Expression of KLRG1 by LCMV-specific CD8 T cells. B6 mice obtained from Harlan Winkelmann (Borchen, Germany) were infected with LCMV (A and B). At the indicated times after infection, PBLs were stained with Db/LCMV GP33-41 tetramers and with monoclonal antibodies specific for CD8 and KLRG1. To stain murine lymphocytes, fluorescein isothiocyanate- or allophycocyanin-labeled anti-CD8 monoclonal antibodies were used that were purchased from BD PharMingen (San Diego, CA). Murine KLRG1 was detected by 2F1 monoclonal antibody (10) purified from hybridoma supernatants by affinity chromatography over protein G-agarose and conjugated with Alexa 488 (Molecular Probes, Eugene, OR) according to the protocol of the manufacturer. The tetramers were prepared by us as described previously (1). The dot plots shown in panel A were gated on CD8 T cells. In panel B, percent tetramer+ cells of CD8 T cells and percent KLRG1+ cells of tetramer+ cells from individual mice are shown.
FIG. 2.
FIG. 2.
Increased KLRG1 expression by LCMV-specific memory CD8 T cells from CD40-deficient mice. B6 (open circles) and B6.CD40−/− (closed circles) mice (obtained from the Jackson Laboratory, Bar Harbor, ME) were infected with LCMV, and at the indicated times after infection, PBLs were stained with Db/LCMV GP33-41 or Db/LCMV NP396-404 tetramers and with monoclonal antibodies specific for CD8 and KLRG1. In panels A and C, percent GP33-41 tetramer+ (A) and NP396-404 tetramer+ (C) cells of CD8 T cells from individual mice are shown. In panels B and D, percent KLRG1+ cells of GP33-41 (B) and of NP396-404 (D) tetramer+ cells from the same mice are depicted. Data are derived from three individual mice (indicated by lines) per group that were analyzed at weeks 1 and 18 after LCMV infection, and relevant P values are indicated.
FIG. 3.
FIG. 3.
Expression of KLRG1 by virus-specific CD8 T cells in humans. (A) PBLs from donors that were known to possess traceable numbers of virus-specific T cells were stained by the indicated tetramers (A2/pp65 for CMV, B8/Nef for HIV, B8/BZLF1 for EBV, and A2/matrix for FLU) and with monoclonal antibodies specific for CD8 and KLRG1. Antibody and tetramer staining was performed as previously described using either heparinized blood, buffy coats, or peripheral blood mononuclear cells isolated from blood by using Ficoll-Hypaque (Amersham Biosciences, Uppsala, Sweden) density centrifugation. Antibodies were obtained from BD PharMingen. Human KLRG1 was detected by Alexa 488-labeled 13A2 monoclonal antibody or by biotinylated or phycoerythrin-labeled 13F12F2 monoclonal antibody (15, 32). Shown are representative dot plots gated on CD8 T cells with percentages of cells present in the different quadrants. (B) Percent KLRG1+ cells of CMV-, HIV-, EBV-, and FLU-specific CD8 T cells identified with the tetramers. Each circle within one particular virus group represents the value from one donor. All donors were analyzed in the chronic (CMV, HIV, and EBV) or postacute (FLU) phase of the infection. KLRG1 expression levels in CMV- and EBV-specific T cells were independent of the HIV status, and FLU-specific T cells were analyzed from HIV-seronegative donors. Peripheral blood mononuclear cells from healthy adult donors (>20 years) were obtained from the Blood Transfusion Center, University Hospital Freiburg, Freiburg, Germany. Further samples were obtained from healthy laboratory workers and from patients chronically infected with HIV, EBV, or CMV, attending clinics in Oxford, United Kingdom. The medical history of each subject was recorded, and blood samples were drawn for serological, virological, and immunological analyses. The study protocol was approved by the local ethics committee.
FIG. 4.
FIG. 4.
Expression of KLRG1 in CD27/CD28 subsets of human CD8 T cells. PBLs from healthy donors were stained with monoclonal antibodies specific for CD8, CD27, CD28, and KLRG1. (A) Histograms on the right display KLRG1 expression gated on the regions R1 to R4 that are indicated in the CD27/CD28 dot plot on the left. A representative staining from one donor is shown. (B) Percent KLRG1+ cells in the indicated CD27/CD28 subsets of CD8 T cells. Symbols represent values from individual donors.
FIG. 5.
FIG. 5.
Coexpression of KLRG1 and CD62L by virus-specific CD8 T cells in humans and mice. (A) P14 TCR-transgenic memory T cells were generated in an adoptive transfer system as described previously (29). Three weeks after LCMV infection, splenic P14 memory cells, identified by the Thy.1 marker, were examined for mouse KLRG1 versus CD62L expression. (B) KLRG1 versus CD62L expression by human FLU tetramer+ CD8 T cells from a healthy donor.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Altman, J. D., P. A. Moss, P. J. Goulder, D. H. Barouch, M. G. McHeyzer-Williams, J. I. Bell, A. J. McMichael, and M. M. Davis. 1996. Phenotypic analysis of antigen-specific T lymphocytes. Science 274:94-96. - PubMed
    1. Anfossi, N., S. H. Robbins, S. Ugolini, P. Georgel, K. Hoebe, C. Bouneaud, C. Ronet, A. Kaser, C. B. DiCioccio, E. Tomasello, R. S. Blumberg, B. Beutler, S. L. Reiner, L. Alexopoulou, O. Lantz, D. H. Raulet, L. Brossay, and E. Vivier. 2004. Expansion and function of CD8+ T cells expressing Ly49 inhibitory receptors specific for MHC class I molecules. J. Immunol. 173:3773-3782. - PubMed
    1. Appay, V., P. R. Dunbar, M. Callan, P. Klenerman, G. M. Gillespie, L. Papagno, G. S. Ogg, A. King, F. Lechner, C. A. Spina, S. Little, D. V. Havlir, D. D. Richman, N. Gruener, G. Pape, A. Waters, P. Easterbrook, M. Salio, V. Cerundolo, A. J. McMichael, and S. L. Rowland-Jones. 2002. Memory CD8+ T cells vary in differentiation phenotype in different persistent virus infections. Nat. Med. 8:379-385. - PubMed
    1. Azuma, M., J. H. Phillips, and L. L. Lanier. 1993. CD28− T lymphocytes. Antigenic and functional properties. J. Immunol. 150:1147-1159. - PubMed
    1. Bachmann, M. F., L. Hunziker, R. M. Zinkernagel, T. Storni, and M. Kopf. 2004. Maintenance of memory CTL responses by T helper cells and CD40-CD40 ligand: antibodies provide the key. Eur. J. Immunol. 34:317-326. - PubMed

Publication types