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IMMUNOBIOLOGY
From the Renal Transplant Unit, Department of Internal
Medicine; Clinical Immunology Laboratory and Department of
Immunobiology, Central Laboratory for Blood Transfusion and Laboratory
for Experimental and Clinical Immunology; and Department of Clinical
Virology, Academic Medical Center; and Department of Immunology, The
Netherlands Cancer Institute, Amsterdam.
During immunosuppression, cytomegalovirus (CMV) can
reactivate and cause serious clinical problems. Normally, abundant
virus replication is suppressed by immune effector mechanisms. To study the interaction between CD8+ T cells and persisting
viruses, frequencies and phenotypes of CMV-specific CD8+ T
cells were determined in healthy individuals and compared to those in
renal transplant recipients. In healthy donors, function of circulating
virus-specific CD8+ T cells, as measured by peptide-induced
interferon The immunologic control of persistent viral
infections, like infections with the family of herpes viruses, requires
the coordinated actions of many cell types.1
CD4+ T cells appear to play a key role in this process
because they orchestrate the various effector arms of the immune
system. In mice it has been shown that the production of neutralizing
antibodies to many viruses critically depends on the availability of
specific CD4+ helper T cells.2,3 In line with
this, we recently demonstrated that in primary human cytomegalovirus
(CMV) infection the emergence of helper T cells precedes the appearance
of virus-specific antibodies.4 CD8+ T cells
eliminate virus-infected cells and are thought to be the major effector
cells in controlling persistent infection. In many instances,
differentiation of CD8+ T cells into competent effector
cells depends on the presence of helper CD4+ T
cells.5,6
On encountering viruses, CD8+ T cells can differentiate
from naive T cells into effector T cells that through cytolysis and secretion of specific cytokines control virus replication and into
memory cells that provide enhanced immunity after renewed contact with
the same pathogen. To study the development of these cells in
clinically relevant situations in humans, various phenotypic markers
have been used that allow separation of the aforementioned subsets.
Naive CD8+ T cells express CD45RA and CD62L and also the
costimulatory receptors CD27 and CD28, whereas memory-type cells
express CD45R0, CD27, and CD28 molecules and low levels of CD62L.
Effector-type cells have a
CD45RA+CD27 Human CMV is a persistent Subjects
For the longitudinal study, 6 HLA-A2+,
CMV-seropositive renal transplant recipients were selected. Heparinized
peripheral blood samples were collected before transplantation and 12 months after transplantation. Peripheral blood mononuclear cells
(PBMCs) were isolated using standard density gradient centrifugation
techniques as described by Rentenaar and colleagues.10 All
patients gave written informed consent and the study was approved by
the local medical ethical committee.
Generation of HLA-A2.1/CMVpp65(NLVPMVATV)
tetrameric complexes
Immunofluorescent staining and flow cytometry Freshly isolated or thawed PBMCs were resuspended in RPMI, containing 10% fetal calf serum (FCS) and antibiotics. Half a million PBMCs were incubated with fluorescent-labeled conjugated monoclonal antibodies (mAbs; concentrations according to manufacturer's instructions) and an appropriate concentration of tetrameric complexes in a small volume for 30 minutes at 4°C, protected from light. To validate specificity of the HLA-A2.1/CMVpp65(NLVPMVATV) a control HLA-A2.1 tetramer containing the HIV-specific SLYNTVATL peptide (a gift from Stefan Kostense, Central Laboratory for Blood Transfusion (CLB), Amsterdam, The Netherlands) was used as negative control. Additional negative controls consisted of HLA-A2.1 CMV-seropositive or
HLA-A2.1+ CMV-seronegative healthy individuals
and renal transplant recipients. Negative controls always showed
tetramer staining of less than 0.01% of total lymphocytes (data not
shown). Cells were washed in phosphate-buffered saline (PBS) containing
0.01% (w/v) NaN3 and 0.5% (w/v) bovine serum albumin
(PBA). For staining with the mouse antihuman CCR7 mAb, a 3-step
staining protocol was performed consisting of incubation with the
anti-CCR7 antibody (a gift from Lijun Wu, Leukosite, Cambridge, MA) for
30 minutes at 4°C protected from light, washing, incubation with
PE-conjugated goat anti-mouse immunoglobulin (Ig; Southern
Biotechnology, Birmingham AL) for 30 minutes at 4°C protected from
light, incubation with 10% (v/v) normal mouse serum (CLB, Amsterdam,
The Netherlands), followed by incubation with directly conjugated mAbs
and tetrameric complexes for 30 minutes at 4°C protected from light.
Analyses consisted of APC-conjugated tetramers and CD8-PerCP (Becton
Dickinson, San Jose, CA) in combinations with either
CD45RA-fluorescein isothiocyanate (FITC; Becton Dickinson) and CD27-PE
(Becton Dickinson), anti-CCR7 and CD45RA-FITC (Becton Dickinson),
CD94-PE (Pharmingen, San Diego, CA), NKB-1-PE (Becton Dickinson),
CD158a-PE, or CD158b-PE (both Immunotech, Marseilles, France). Analysis
of cells for the expression of cell surface markers was performed
using a FACS Calibur flow cytometer and Cellquest software (Becton
Dickinson). In addition, CCR7 expression on CD8+
T cells was analyzed by flow cytometry, using goat antimouse Ig-FITC
(CLB) in conjunction with either CD8-PerCP (both Becton Dickinson), CD45RA-PE, CD27-biotin (both CLB), and streptavidin-APC (Pharmingen) or CD28-PE and CD45R0-APC (both Becton Dickinson).
Determination of CMV-specific CD4+ and CD8+ T cells by intracellular cytokine staining The CMV-specific CD4+ and CD8+ T-cell frequencies were determined essentially according to the method described by Waldrop and coworkers13 and Kern and colleagues,14 respectively. Briefly, 106 freshly isolated PBMCs were incubated for 6 hours in the presence of either CMV antigen (Biowhittaker, Wokingham, United Kingdom, 60 µL/mL), control antigen (Biowhittaker, 60 µL/mL, negative control) (determination of CMV-specific CD4+ T cells), Staphylococcus aureus enterotoxin B (SEB, ICN/Fluka, 2 µg/mL, positive control), the HLA-A2-binding CMV peptide or the HLA-B7-binding CMV peptide (negative control), or the HLA-A2-binding HIV peptide (negative control) (see below, final concentration of 10 µg/mL, determination of CMV-specific CD8+ T cells). CD28 mAb (clone 15E8 CLB) was added as 3 µg/mL (final concentration) in a final volume of 2 mL/tube RPMI 1640 (Gibco, Paisley, United Kingdom) containing 10% heat inactivated FCS (Integro, Zaandam, The Netherlands), penicillin, and streptomycin. For the final 5 hours of culture, brefeldin A (Sigma) was added to the culture in a final concentration of 10 µg/mL. Cells were transferred to FACS tubes, fixed in 2 mL/tube FACS lysing solution (Becton Dickinson), permeabilized in 0.5 mL/tube FACS permeabilizing solution followed by (intracellular) staining with IFN- -FITC (Becton
Dickinson) and CD69-PE (Becton Dickinson) and CD4-APC (Becton
Dickinson) or CD8-APC (21C008AX, clone UCHT-4, Imgen, ITK, Uithoorn,
The Netherlands). Cells were washed in PBA and refixed in
Cellfix (Becton Dickinson) until flow cytometric analysis the following day. Flow cytometric analysis was performed using a FACS Calibur equipped with a 488-nm argon ion laser and a 635-nm red diode laser.
Data files containing 50 000 events positive for CD4-APC or CD8-APC
fluorescence within a lymphocyte gate were saved. Frequencies of
CD69+IFN-+ cells
within the CD4+ or CD8+
lymphocyte gate were determined using Cellquest software (Becton Dickinson) and designated CMV-specific CD4+ or
CD8+ T-cell frequencies, respectively. Negative
controls showed less than 0.05% of
CD69+IFN-+ cells (data
not shown).
Peptides The HLA-A2 binding CMVpp65-derived peptide NLVPMVATV and the HLA-B7 binding CMVpp65-derived peptide TPRVTGGA were purchased from the IHB-LUMC peptide synthesis library facility (Leiden, The Netherlands). The HIV Gag p17-derived peptide SLYNTVATL was kindly provided by Stefan Kostense (CLB, Amsterdam, The Netherlands). The peptides were generated by standard Fmoc techniques and purified by ether precipitation and HPLC techniques. The peptides were dissolved in dimethylsulfoxide (DMSO; Merck, Darmstadt, Germany) in a concentration of 5 mg/mL.Intracellular granzyme B and perforin staining Intracellular granzyme B and perforin staining was performed as described previously.16 In short, half a million PBMCs were stained with fluorescent-labeled conjugated mAbs to CD8 (Becton Dickinson), CD27 (Becton Dickinson), and CMV-tetrameric complexes, washed once with PBA, then fixed with 50 µL buffered formaldehyde acetone solution and subsequently permeabilized by washing with 0.1% saponine 50 mM D-glucose. Cells were then incubated with antigranzyme B (CLB) and antiperforin antibodies (Hölzel Diagnostika, Köln, Germany) according to manufacturers' instructions. Flow cytometric analysis was performed immediately.CMV-polymerase chain reaction Quantitative polymerase chain reaction (PCR) was performed in EDTA whole blood samples as described for plasma or serum.4Viral culture Viral culture was done by cocultivation of urine and human diploid fibroblasts. Microscopic examination for the appearance of CMV-specific cytopathologic effects was performed.Anti-CMV IgG Anti-CMV IgG was determined in serum using the AxSYM microparticle enzyme immunoassay (Abbott, Abbott Park, IL) according to the manufacturer's instructions. Measurements were calibrated relative to a standard serum. Results are expressed as a ratio of the measurement to a standard serum (IgM).Statistical analysis Between-group analysis was performed using the nonparametric Mann-Whitney test. Within-group analysis was performed using the Wilcoxon signed rank test. Two-sided testing was done; P values lower than .05 were considered statistically significant.
Correlations of CMV-specific CD4+ and CD8+ T-cell frequencies in healthy individuals As previously demonstrated by us and others, CMV-specific CD4+ T cells can readily be detected in the circulation of healthy virus carriers using CMV antigen-induced IFN- production as
read-out (Figure 1A). Likewise,
CD8+ T cells could be visualized using the immunodominant
CMV peptide in HLA-A2+ donors (Figure 1B). Frequencies of
IFN--producing cells within the CD8+ subset ranged
between 0.18% and 0.80%. (Figure 2A).
To directly visualize CMV-specific T cells, tetrameric
HLA-A2.1/NLVPMVATV complexes were generated. These complexes bound
0.54% to 3.77% of CD8+ T cells (Figures 1C, 2B).
Accordingly, only on average 20% of the peptide-specific T cells in
these chronic virus carriers were able to secrete IFN- in this
short-term activation assay. Interestingly, in line with studies in
mice, CD4 helper T-cell frequencies correlated with the percentage of
CD8+ T cells secreting IFN- after peptide stimulation
(Figure 1D, r = 0.7982, P < .05) but not with the
amount of specific tetramer binding cells (data not shown).
Cell surface phenotype of CMV-specific CD8+ T cells in healthy individuals On the basis of functional similarities of the subsets defined by CD27 and CCR7,7,9 one would expect that these molecules are being expressed in a concordant way. The localization of CCR7 cells in CD8+ T-cell subsets defined by
CD27 and CD45RA expression was determined (Figure
3C). Confirming recently published
data,17 most (mean 97.0%, n = 4 healthy donors)
effector-type CD45RA+CD27 T
cells7 lacked CCR7. Additional analysis showed that most CCR7 T cells also lacked CD28 and CD45R0 molecules
(Figure 3D). However, an appreciable number of CCR7 T
cells appeared to be contained within the
CD27+CD28+CD45R0+ population (mean
33.1%). CCR7 T cells were virtually absent from the
naïve T-cell subset (mean 1.5%).
To test whether in antigen-specific T cells expression of the various
markers is comparable to that in the total population, CMV-specific T
cells (Figure 4A) were analyzed for CD27,
CCR7, and CD45RA coexpression. In all donors, CMV-specific
CD8+ T cells were of the TEM
type,9 that is, lacked expression of CCR7 (Figure 4B).
Remarkably, however, with respect to the expression of CD27 and CD45RA
molecules heterogeneous phenotypes were found. In most healthy donors,
CMV-specific T cells appeared to be predominantly memory cells, that
is, CD27+CD45RA
Cytolytic mediators and natural killer cell receptors in CMV-specific T cells The absence of CD27 corresponds within the expression of cytolytic mediators such as perforin and granzyme B.7 To establish if this phenomenon holds for virus-specific T cells, CMV-specific CD8+ T cells of a healthy donor with a predominant effector phenotype, and cells of one healthy donor with a predominant memory phenotype were analyzed for granzyme B expression by flow cytometry. Indeed, a high percentage of antigen-specific effector cells, that is, CD27CCR7CD45RA+, contained
granzyme B, whereas only low numbers of
CD27+CCR7CD45RA cells contained
the molecule (Figure 4D-E).
In analogy with findings on CD28
Frequencies and properties of CMV-specific CD8+ T cells in renal transplant recipients Frequencies of CMV-specific helper CD4+ T cells as measured by antigen-induced IFN- secretion did not differ
significantly between healthy donors and recipients of renal
transplants who were taking immunosuppressive drugs for on
average 36.5 months (median 0.20 versus 0.47%, P = .53,
data not shown and Rentenaar et al4). However,
the frequencies of virus-specific CD8+ T cells analyzed
either by peptide-induced IFN- secretion or by direct visualization
with HLA-A2.1-NLVPMVATV tetramers were significantly higher in the
transplant recipients (P = .0082 respectively .0056, Figure 2A-B). Similar to CD8+ T cells in healthy donors
approximately 20% of the antigen-specific CD8+ T cells
were able to secrete IFN- in 6-hour stimulation assays (P = .558 between control individuals and transplant
recipients, data not shown). In renal transplant recipients no
significant correlation was found between helper cell frequencies and
either IFN--producing or number of CMV-specific CD8+ T
cells (data not shown).
Phenotypic analyses showed that, in accordance with data obtained in
healthy donors, CMV-specific CD8+ T cells in renal
transplant recipients were CCR7
Repetitive antigenic stimulation induces loss of CD27 The transsectional findings and the data of Gillespie21 suggested that immunosuppressive therapy altered the CMV-specific CD8+ T-cell compartment in a quantitative and qualitative fashion. To test this directly, we investigated the effect of immunosuppressive therapy on the phenotype of CMV-specific CD8+ T cells in patients before and 12 months after transplantation. Mean frequencies of CMV-specific CD8+ T cells rose from 2.08% to 3.31% during this period (P = .313, not significant). Moreover, in 5 of 6 patients the percentage of CD27 effector cells increased at the
expense of the CD27+ memory cells. Interestingly, this
change was most dramatic in a patient who had continuous positive urine
cultures for CMV after transplantation (Figure
7A-B).
The data presented in this paper establish a number of key
features of CD8+ T cells involved in the control of
persistent viruses, specifically CMV. CD4+ T cells are
believed to be instrumental in the initiation of CD8+
T-cell expansion via the stimulation of dendritic
cells,22,23 but their role in maintaining adequate numbers
and function of specific CD8+ T cells is less well
understood. Zajac and colleagues5 observed that in
CD4 The CMV-specific CD8+ T cells can be typified as
TEM cells, but based on expression of CD27 a further
subdivision in functional subpopulations can be made. We would prefer
to reserve the annotation "effector" for the CD27 When CD8+ T cells progress through the proposed naive, memory, and effector stages,32 expression of NKRs, both of the Ig and C-type lectin type, coordinately increases.19 Inhibitory NKRs may set thresholds for activation precluding that any type of cell with low antigen expression would be targeted by circulating effector cytotoxic T lymphocytes (CTLs). Huard and Karlsson20 suggested that repeated exposure to specific antigens would increase expression of NKRs on specific T cells. Our data, however, unequivocally demonstrate that NKRs are barely if all expressed on T cells specific for CMV. The reason for this apparent paradox is unclear at this moment. Probably, antigens that persistently stimulate the immune system differ in their way of being presented and the context of presentation may have both direct and indirect effects on the expression of activation-regulating receptors. Elegant work from several groups has shown that CMV contains several genes that influence immune recognition.33-36 In this perspective it will be of interest to determine if other persisting viruses recruit memory and effector cells that also lack NKRs. In any case the data infer that control of CMV replication might be efficient because of lack of interference by NKRs in conjunction with high frequencies of CMV-specific effector cells. The CMV-specific T cells have been detected in CD45R0high,
CD45RAhigh, and CD28 Our data infer that despite effective immunosuppression of allospecific
T-cell-mediated graft rejection the immune system is able to mount an
adaptive response to persistent viruses such as CMV. In most healthy
individuals, an equilibrium is achieved with a predominance of
virus-controlling memory-type CD8+ T cells. However, in
response to viral and host factors, CD8 responses may be shifted toward
effector-type cells. Indeed, in a number of our normal donors
effector-type cells predominated. These individuals were healthy and
did not have signs of viral replication in vivo. Interestingly, in
elderly individuals the proportion of CD27
The authors thank the patients and volunteers for their blood donations, Dr F. Bemelman for assistance in collecting patient material, Frank van Diepen for excellent technical assistance, Dr R. Boom for development of the CMV-PCR, technicians from the Department of Clinical Virology for performing CMV-PCRs and cultures, and Drs Frank Miedema, Debbie van Baarle, and Michiel Betjes for critical reading of the manuscript. Dr Lijun Wu is thanked for providing the anti-CCR7 antibody and Dr Stefan Kostense for providing the HLA-A2/HIV-SLYNTVATL tetramers and peptide.
Submitted October 30, 2000; accepted April 4, 2001.
Supported by grants from the Dutch Kidney Foundation, C98-1724 (L.E.G.) and C95-1455 (R.J.R.).
L.E.G. and R.J.R. contributed equally to this work.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Laila E. Gamadia, Clinical Immunology Laboratory G1-109, Academic Medical Center, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands; e-mail: l.e.gamadia{at}amc.uva.nl.
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P. Romero, A. Zippelius, I. Kurth, M. J. Pittet, C. Touvrey, E. M. Iancu, P. Corthesy, E. Devevre, D. E. Speiser, and N. Rufer Four Functionally Distinct Populations of Human Effector-Memory CD8+ T Lymphocytes J. Immunol., April 1, 2007; 178(7): 4112 - 4119. [Abstract] [Full Text] [PDF]
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E. M. M. van Leeuwen, J. J. Koning, E. B. M. Remmerswaal, D. van Baarle, R. A. W. van Lier, and I. J. M. ten Berge Differential Usage of Cellular Niches by Cytomegalovirus versus EBV- and Influenza Virus-Specific CD8+ T Cells J. Immunol., October 15, 2006; 177(8): 4998 - 5005. [Abstract] [Full Text] [PDF]
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D. Lilleri, G. Gerna, C. Fornara, L. Lozza, R. Maccario, and F. Locatelli Prospective simultaneous quantification of human cytomegalovirus-specific CD4+ and CD8+ T-cell reconstitution in young recipients of allogeneic hematopoietic stem cell transplants Blood, August 15, 2006; 108(4): 1406 - 1412. [Abstract] [Full Text] [PDF]
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M. Guma, M. Budt, A. Saez, T. Brckalo, H. Hengel, A. Angulo, and M. Lopez-Botet Expansion of CD94/NKG2C+ NK cells in response to human cytomegalovirus-infected fibroblasts Blood, May 1, 2006; 107(9): 3624 - 3631. [Abstract] [Full Text] [PDF]
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O. A. Shlobin, E. E. West, N. Lechtzin, S. M. Miller, M. Borja, J. B. Orens, L. K. Dropulic, and J. F. McDyer Persistent Cytomegalovirus-Specific Memory Responses in the Lung Allograft and Blood following Primary Infection in Lung Transplant Recipients J. Immunol., February 15, 2006; 176(4): 2625 - 2634. [Abstract] [Full Text] [PDF]
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Y. Chalandon, S. Degermann, J. Villard, L. Arlettaz, L. Kaiser, S. Vischer, S. Walter, M. H. M. Heemskerk, R. A. W. van Lier, C. Helg, et al. Pretransplantation CMV-specific T cells protect recipients of T-cell-depleted grafts against CMV-related complications Blood, January 1, 2006; 107(1): 389 - 396. [Abstract] [Full Text] [PDF]
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E. M. M. van Leeuwen, G. J. de Bree, E. B. M. Remmerswaal, S.-L. Yong, K. Tesselaar, I. J. M. t. Berge, and R. A. W. van Lier IL-7 receptor {alpha} chain expression distinguishes functional subsets of virus-specific human CD8+ T cells Blood, September 15, 2005; 106(6): 2091 - 2098. [Abstract] [Full Text] [PDF]
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M. Guma, A. Angulo, C. Vilches, N. Gomez-Lozano, N. Malats, and M. Lopez-Botet Imprint of human cytomegalovirus infection on the NK cell receptor repertoire Blood, December 1, 2004; 104(12): 3664 - 3671. [Abstract] [Full Text] [PDF]
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E. M. Aandahl, M. F. Quigley, W. J. Moretto, M. Moll, V. D. Gonzalez, A. Sonnerborg, S. Lindback, F. M. Hecht, S. G. Deeks, M. G. Rosenberg, et al. Expansion of CD7low and CD7negative CD8 T-cell effector subsets in HIV-1 infection: correlation with antigenic load and reversion by antiretroviral treatment Blood, December 1, 2004; 104(12): 3672 - 3678. [Abstract] [Full Text] [PDF]
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E. M. M. van Leeuwen, E. B. M. Remmerswaal, M. T. M. Vossen, A. T. Rowshani, P. M. E. Wertheim-van Dillen, R. A. W. van Lier, and I. J. M. ten Berge Emergence of a CD4+CD28- Granzyme B+, Cytomegalovirus-Specific T Cell Subset after Recovery of Primary Cytomegalovirus Infection J. Immunol., August 1, 2004; 173(3): 1834 - 1841. [Abstract] [Full Text] [PDF]
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E. R. Piriou, K. van Dort, N. M. Nanlohy, F. Miedema, M. H. van Oers, and D. van Baarle Altered EBV Viral Load Setpoint after HIV Seroconversion Is in Accordance with Lack of Predictive Value of EBV Load for the Occurrence of AIDS-Related Non-Hodgkin Lymphoma J. Immunol., June 1, 2004; 172(11): 6931 - 6937. [Abstract] [Full Text] [PDF]
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L. E. Gamadia, E. M. M. van Leeuwen, E. B. M. Remmerswaal, S.-L. Yong, S. Surachno, P. M. E. Wertheim-van Dillen, I. J. M. ten Berge, and R. A. W. van Lier The Size and Phenotype of Virus-Specific T Cell Populations Is Determined by Repetitive Antigenic Stimulation and Environmental Cytokines J. Immunol., May 15, 2004; 172(10): 6107 - 6114. [Abstract] [Full Text] [PDF]
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E. Mallard, F. Vernel-Pauillac, T. Velu, F. Lehmann, J.-P. Abastado, M. Salcedo, and N. Bercovici IL-2 Production by Virus- and Tumor-Specific Human CD8 T Cells Is Determined by Their Fine Specificity J. Immunol., March 15, 2004; 172(6): 3963 - 3970. [Abstract] [Full Text] [PDF]
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M. K. Gandhi, M. R. Wills, G. Okecha, E. K. Day, R. Hicks, R. E. Marcus, J. G. P. Sissons, and A. J. Carmichael Late diversification in the clonal composition of human cytomegalovirus-specific CD8+ T cells following allogeneic hemopoietic stem cell transplantation Blood, November 1, 2003; 102(9): 3427 - 3438. [Abstract] [Full Text] [PDF]
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N. Rufer, A. Zippelius, P. Batard, M. J. Pittet, I. Kurth, P. Corthesy, J.-C. Cerottini, S. Leyvraz, E. Roosnek, M. Nabholz, et al. Ex vivo characterization of human CD8+ T subsets with distinct replicative history and partial effector functions Blood, September 1, 2003; 102(5): 1779 - 1787. [Abstract] [Full Text] [PDF]
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D. Sauce, N. Rufer, P. Mercier, M. Bodinier, J.-P. Remy-Martin, A. Duperrier, C. Ferrand, P. Herve, P. Romero, F. Lang, et al. Retrovirus-mediated gene transfer in polyclonal T cells results in lower apoptosis and enhanced ex vivo cell expansion of CMV-reactive CD8 T cells as compared with EBV-reactive CD8 T cells Blood, August 15, 2003; 102(4): 1241 - 1248. [Abstract] [Full Text] [PDF]
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W. J. M. Mackus, F. N. J. Frakking, A. Grummels, L. E. Gamadia, G. J. de Bree, D. Hamann, R. A. W. van Lier, and M. H. J. van Oers Expansion of CMV-specific CD8+CD45RA+CD27- T cells in B-cell chronic lymphocytic leukemia Blood, August 1, 2003; 102(3): 1057 - 1063. [Abstract] [Full Text] [PDF]
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T. W. Kuijpers, M. T. Vossen, M.-R. Gent, J.-C. Davin, M. T. Roos, P. M. Wertheim-van Dillen, J. F. Weel, P. A. Baars, and R. A. van Lier Frequencies of Circulating Cytolytic, CD45RA+CD27-, CD8+ T Lymphocytes Depend on Infection with CMV J. Immunol., April 15, 2003; 170(8): 4342 - 4348. [Abstract] [Full Text] [PDF]
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L. E. Gamadia, E. B. M. Remmerswaal, J. F. Weel, F. Bemelman, R. A. W. van Lier, and I. J. M. Ten Berge Primary immune responses to human CMV: a critical role for IFN-gamma -producing CD4+ T cells in protection against CMV disease Blood, April 1, 2003; 101(7): 2686 - 2692. [Abstract] [Full Text] [PDF]
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E. M. van Leeuwen, L. E. Gamadia, P. A. Baars, E. B. Remmerswaal, I. J. ten Berge, and R. A. van Lier Proliferation Requirements of Cytomegalovirus-Specific, Effector-Type Human CD8+ T Cells J. Immunol., November 15, 2002; 169(10): 5838 - 5843. [Abstract] [Full Text] [PDF]
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Y.-J. Kim, R. R. Brutkiewicz, and H. E. Broxmeyer Role of 4-1BB (CD137) in the functional activation of cord blood CD28-CD8+ T cells Blood, October 16, 2002; 100(9): 3253 - 3260. [Abstract] [Full Text] [PDF]
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N. Khan, N. Shariff, M. Cobbold, R. Bruton, J. A. Ainsworth, A. J. Sinclair, L. Nayak, and P. A. H. Moss Cytomegalovirus Seropositivity Drives the CD8 T Cell Repertoire Toward Greater Clonality in Healthy Elderly Individuals J. Immunol., August 15, 2002; 169(4): 1984 - 1992. [Abstract] [Full Text] [PDF]
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V. Monsurro, D. Nagorsen, E. Wang, M. Provenzano, M. E. Dudley, S. A. Rosenberg, and F. M. Marincola Functional Heterogeneity of Vaccine-Induced CD8+ T Cells J. Immunol., June 1, 2002; 168(11): 5933 - 5942. [Abstract] [Full Text] [PDF]
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C. Gioia, C. Agrati, R. Casetti, C. Cairo, G. Borsellino, L. Battistini, G. Mancino, D. Goletti, V. Colizzi, L. P. Pucillo, et al. Lack of CD27-CD45RA-V{gamma}9V{delta}2+ T Cell Effectors in Immunocompromised Hosts and During Active Pulmonary Tuberculosis J. Immunol., February 1, 2002; 168(3): 1484 - 1489. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
-herpesvirus that is present in
approximately 50% of the adult population. Latent infection is asymptomatic in healthy individuals but can cause serious disease in
immunocompromised individuals. Severely impaired T-cell function leads
to viral reactivation and morbidity due to cytopathic effects of
uncontrolled viral replication, and CMV reactivation coincides with
higher levels of immunosuppressive therapy. Interestingly, however,
most renal transplant recipients who receive medication for adequate
suppression of the allogeneic immune response do not suffer from
episodes of clinically apparent CMV reactivation and therefore appear
to efficiently control overt virus replication. Recent technologic
developments such as the generation of tetrameric major
histocompatibility complex (MHC)-peptide complexes and peptide-induced interferon 
) and CMV-seropositive
HLA-A2+ renal transplant recipients (
). Frequencies of
these CMV-specific CD8+ T cells were higher in renal
transplant recipients (Mann-Whitney, P = .0082). (B)
Frequencies of NLVPMVATV-HLA-A2.1 tetramer binding CD8+ T
cells (y-axis, percentage of CD8+ T cells) in
CMV-seropositive HLA-A2+ control individuals without
immunosuppression (
, y-axis, percentage of tetramer-positive CD8+ T
cells) from donors with an effector (left bar) or memory (right bar)
phenotype.
