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IMMUNOBIOLOGY
From the Department of Infectious Diseases and
Microbiology, Graduate School of Public Health, and the Department of
Pathology, School of Medicine, University of Pittsburgh, Pennsylvania;
the Department of Microbiology and Immunology, University of Oklahoma
Health Sciences Center, Oklahoma City; and the Immunex Corporation,
Seattle, WA.
Human herpesvirus 8 (HHV-8; Kaposi sarcoma-associated
herpesvirus)-specific cytotoxic T-lymphocyte (CTL) and interferon- Human herpesvirus 8 (HHV-8; also termed Kaposi
sarcoma-associated herpesvirus) is a gammaherpesvirus that is
considered to be the causative agent of Kaposi sarcoma (KS), body
cavity B-cell lymphoma, and multicentric Castleman
disease.1 T-cell immunity is thought to play an important
role in control of HHV-8 infection and these associated
diseases.2-5 In this regard, we2,3 and
others4 have demonstrated that major histocompatibility complex (MHC) class I-restricted, CD8+ cytotoxic T
lymphocytes (CTLs), and interferon Support for the potential role of CD8+ T-cell immunity in
control of HHV-8 infection comes from several studies on immune control of acute and persistent infections with 2 other
gammaherpesviruses, specifically, Epstein-Barr virus (EBV) and
murine herpesvirus 68 (MHV-68).6-10 Although EBV latency
proteins can induce strong CD8+ T-cell
responses,11 lytic cycle proteins are considered as major
CD8+ T-cell targets in both acute and persistent EBV
infections.12 Immunodominant epitopes restricted to
different HLA class I alleles, which are important for our
understanding of viral immunopathogenesis and development of
appropriate vaccines,13 have been determined for peptides
derived from EBV structural proteins gp350 and gp85, and
immediate-early (IE) and early proteins such as BZLF1, BRLF1, and
BMLF1.6-9 Compared to latency proteins, the frequency of CD8+ T lymphocytes specific for peptides derived from lytic
cycle proteins is higher in PBMCs of healthy virus
carriers.14 CD8+ CTLs have also been
demonstrated to secrete IFN- In MHV-68 infection, lytic cycle proteins also induce CD8+
T cell responses.10,18,19 T cells specific for peptides
derived from lytic cycle proteins dominate the acute phase of MHV-68
infection.10,19 Moreover, vaccination with immunodominant
CD8+ T-cell epitopes derived from lytic cycle proteins of
MHV-68 significantly reduces viral titers and the level of infected
cells during acute infection.20
We have tested CD8+ T-cell reactivity to peptides derived
from 5 HHV-8 lytic cycle proteins based on a prediction model for the
HLA A*0201 motif in HHV-8-infected subjects to determine
immunodominant epitopes to the virus. We chose HLA A*0201 because it is
prevalent in a large percentage of the white population and it is the
most extensively studied HLA class I antigen.21 We found
that one peptide derived from the glycoprotein B (gB) homolog
was immunogenic for CD8+ T cells from HHV-8+,
HLA A*0201 individuals, particularly when presented by autologous, mature dendritic cells (DCs). CD8+ T-cell responses to this
HHV-8 lytic cycle peptide may be important in the control of acute and
persistent HHV-8 infection and in vaccine design.
Study subjects and detection of HHV-8 infection
Detection of HHV-8 serum antibody specific for lytic antigens and HLA
molecular typing were done as described elsewhere.2,3,23 HHV-8 DNA was detected in PBMCs by DNA hybridization to HHV-8-specific polymerase chain reaction (PCR) products. PCR analyses were performed using primers that amplified a 233-base pair (bp) fragment of the
minor capsid gene encoded by open reading frame (ORF)
2624,25 as described previously.3 PCR products
were separated on 2% agarose gels, transferred to Nytran
membranes and hybridized with an internal oligonucleotide probe labeled
with 32P. The sequence of oligonucleotide probe was
TGCAGCAGCTGTTGGTGTACCACAT. PCR products that hybridized to the probe
were detected by image analysis with an ImageQuant PhosphorImager
(Molecular Dynamics, Sunnyvale, CA). This procedure has a sensitivity
of detection of 10 or more copies of HHV-8 DNA from
1 × 106 PBMCs. We have also found that PBMCs from HHV-8
seronegative controls are negative for HHV-8 DNA, whereas a cell line
persistently infected with HHV-8 is consistently positive by this procedure.
Synthetic peptides
T2 cell line HLA A*0201 stabilization assay A peptide-induced stabilization assay of the HLA A*0201 molecule expressed by the T2 cell line was performed using a method described elsewhere.29,30 T2 cells are a transporter associated with antigen processing (TAP)-deficient cell line that has low levels of HLA A*0201 molecules expressed on the cell surface and impaired endogenous peptide presentation.31 The T2 stabilization assay determines the conformation of HLA A*0201 molecules when exogenous synthetic peptides and 2-microglobulin
(2M) are delivered to the T2 cells. Two control peptides
in addition to the 25 HHV-8 lytic cycle peptides were tested. A peptide
derived from HIV-1 p24 protein (Gag p24263-272; KRWIILGLNK;
single-letter amino acid codes), which is HLA
B27-restricted,32 served as a negative control; a peptide
representing an HLA A*0201 epitope from the influenza A virus matrix
protein (M158-66; GILGFVFTL)33 served as a
positive control. The 5 × 105 T2 cells (gift from Dr
Russell Salter, University of Pittsburgh) were incubated at 37°C and
5% CO2 overnight in the presence of various concentrations
of peptide in RPMI 1640 medium (Life Technologies, Gaithersburg, MD)
supplemented with 10% heat-inactivated fetal calf serum (Hyclone,
Logan, UT) and 1 µM 2M (Sigma, St Louis, MO). The
cells were washed twice with cold phosphate-buffered saline (PBS) and
incubated for 30 minutes at 4°C with monoclonal antibody (mAb)
specific for the conformationally changed HLA class I molecule (BB7.2;
American Type Culture Collection, Manassas, VA). The cells were then
washed twice with PBS and incubated with fluorescein
isothiocyanate-conjugated IgG F(ab')2 (Organon-Teknika, Durham, NC) for 30 minutes at 4°C. The cells were washed twice with
cold PBS and fixed with 1% paraformaldehyde (Polysciences, Warrington,
PA). The labeled cells were analyzed by flow cytometry (Elite; Beckman
Coulter, Fullerton, CA). The positive binding capacity cutoff was
determined by the mean + 3 SD of the ratio for HIV-1
p24263-272, which was calculated as mean fluorescence intensity (MFI) of the test peptides compared to the MFI of T2 cells
without peptides added. The mean ± SD value for HIV-1
p24263-272 binding was 0.93 ± 0.18, giving a cutoff
value for positive binding of 1.5.
Single-cell IFN-  production was done
using standard methods described previously,3 with minor modifications. Frozen PBMCs were thawed and incubated overnight at
1 × 105 cells with peptides at a concentration of 50 µg/mL in nitrocellulose-bottom, 96-well plates (Millipore, Bedford,
MA). An immunodominant peptide derived from an EBV lytic cycle protein
(BMLF1280-288; GLCTLVAML),6 the influenza
matrix peptide M158-66, and HIV-1 p24 Gag peptide (p24151-159; TLNAWVKVV)34 and
phorbol 12-myristrate 13-acetate (1 ng/mL)-ionomycin (1 µM;
PMA-iono) were used as positive controls. A plasmid expressing the ORF
for gB (ORF8) was cloned into the vaccinia virus shuttle vector, pSC11,
as previously described,2,3 and used for expression of the
native form of gB protein (designated VgB). VSC11 not containing the
HHV-8 plasmid was used as the control for background, vaccinia virus
gene expression.
After overnight stimulation with the antigens, the plates were washed
and processed, and the number of spots counted with a dissecting
microscope.3 The results are given as the number of spots
per 106 PBMCs stimulated by the viral peptides or
autologous B lymphocyte cell lines (BLCLs) infected with VgB, minus the
number of spots per 106 PBMCs stimulated by the medium
control or BLCLs infected with the VSC11 vector, respectively. The mean
(± SE) number of IFN- In certain experiments, the peptides were presented to PBMCs by autologous DCs derived from CD14+ blood monocytes that were positively selected using CD14 immunomagnet microbeads (Miltenyi Biotec, Auburn, CA). The CD14+ cells were resuspended at 1 × 106 cells/mL in AIM-V medium containing 1000 U/mL each of recombinant human interleukin (IL)-4 and human granulocyte-monocyte colony-stimulating factor (GM-CSF). The cells were cultured for 7 days with fresh IL-4 and GM-CSF added at 2-day intervals as previously described.34 The cells were then treated with human CD40 ligand trimer (CD40L) at 1 µg/mL (Immunex, Seattle, WA) for 2 days to augment phenotypic and functional maturation. These DCs have a mature phenotype as determined by staining with fluorescent dye-conjugated mAb and analyzed by FACS as previously described.34 The mature DCs were loaded with peptide at 10 µg/mL for 2 hours in AIM V at room temperature, washed 3 times and counted with a light microscope. The peptide-loaded mature DCs were then used as stimulators (5 × 103) for autologous PBMCs (5 × 104) in triplicate in the single-cell enzyme immunoassay overnight at 37°C (termed zero week stimulation, or S0). To examine longer-term effects of stimulation of T cells by DCs, parallel DC-PBMC mixtures were cultured with IL-2 at 100 U/mL for 7 days, harvested, and restimulated (1 × 105 cells) overnight at 37°C with either untreated or peptide loaded, autologous PBMCs (1 × 104) in triplicate in nitrocellulose-bottom microwells in the single-cell enzyme immunoassay (termed 1-week stimulation, or S1). Stimulation of PBMCs with DCs without peptide was included as a control for the S0 and S1 cultures. CTL assay The bulk lysis CTL assay was performed as described previously using 3 effector-to-target (E/T) ratios,2,3 except for the preparation of target cells. On day 13, autologous BLCLs infected with the VgB or VSC11 control vectors, or T2 cells pulsed with the viral peptides (50 µg/mL) and B2M, were labeled with 51Cr (100 µCi/mL [3.7 MBq] Na2[51Cr]O4, Dupont NEN, Boston, MA) and incubated overnight. On day 14, 51Cr release was determined in a gamma counter. The data are presented as percent virus-specific lysis, which equals percent specific lysis against the HHV-8 antigen-expressing targets minus percent specific lysis against the VSC11 control antigen-expressing targets. Mean (± SE) background lysis of the uninfected and VSC11-infected targets was 0 (± 1%) for lysis at 3 E/T ratios (n = 9). The cutoff value for positive lysis was 10% based on anti-HHV-8 CTL lysis in HHV-8-seronegative individuals.2Establishment of CD8+ T-cell lines For the establishment of CD8+ T-cell lines, T2 cells pulsed with the gB492-500 peptide and2M
were used as stimulators and plated into 96-well U-bottom plates at a
concentration of 5 × 104 in RPMI 1640 medium,
supplemented with 15% fetal calf serum, IL-2 (100 U/mL; Chiron,
Emeryville, CA), and anti-CD3 mAb (12-F6, 10 ng/mL; gift from Dr
Johnson Wang, Massachusetts General Hospital, Boston, MA). Frozen PBMCs
were thawed and plated at 50 cells/well in 96-microwell U-bottom
plates. Then, 50 µL gamma-irradiated (4000 rad) allogeneic PBMCs were
plated at a concentration of 1 × 106/mL as feeder cells
to enhance T-cell growth. The cultures were incubated at 37°C in 5%
CO2 for 2 to 3 weeks. Primary screening was performed when
large cell pellets were visually evident. The cell lines were
approximately 70% CD8+ by mAb staining. Radioactively
labeled T2 cells loaded with the corresponding peptides and
B2M were used as targets. Wells positive for CTL activity
were then transferred to 48-well plates and used for the single-cell
IFN- production assay and T-cell phenotyping. T-cell subsets were
phenotyped by flow cytometry (XL; Beckman Coulter) after staining with
mAbs specific for CD3, CD4, and CD8 (Becton Dickinson; Mountain
View, CA).
Selection of potential CD8+ T-cell epitopes of HHV-8 lytic cycle proteins for binding to HLA A*0201 To identify potential HLA A*0201-restricted epitopes within HHV-8 lytic cycle proteins, amino acid sequences of the 5 HHV-8 proteins were analyzed based on a prediction model for the HLA A*0201 binding motif.27,28 This prediction model allows location and ranking of 9-mer peptides that contain putative peptide-binding motifs for HLA class I molecules, based on an estimation of the half-time dissociation (T1/2 dis) of the HLA-peptide complex. The 5 highest predicted binding scores for 9-mer peptides derived from each of the 5 HHV-8 lytic cycle proteins are listed in Table 1.We tested the 25 peptides for actual HLA A*0201 binding capacity using the T2 cell-binding assay. Thirteen of the 25 peptides bound to HLA A*0201 in a concentration-dependent manner (Table 1). The positive control M158-66 peptide also bound, whereas no binding was associated with the negative control p24 Gag263-272 peptide at any concentration tested. Characterization of HHV-8 peptides for stimulation of peripheral blood, CD8+ T cells from HHV-8 seropositive, healthy persons We next characterized CD8+ T-cell reactivity to the HHV-8 peptides using a single-cell IFN- production assay in the 3 groups of HIV-1 , healthy subjects. Even though 12 of the
peptides did not bind to HLA A*0201 based on our T2 cell-binding assay,
we chose to screen all 25 peptides for T-cell activation in the event
that this assay was more sensitive than the binding assay for detecting peptide antigenicity. We found that PBMCs from 2 of the 14 group A
persons, who were HHV-8+ and HLA A*0201-genotypic,
produced IFN- (subject A27 = 50 and subject A29 = 240
spots/106 PBMCs) in response to one of the 13 HHV-8
peptides that bound to HLA A*0201, that is, gB492-500
(LMWYELSKI), and to none of the 12 peptides that did not bind (data not
shown). No HHV-8 peptide-specific, IFN- production was noted in
PBMCs from either the 5 group B subjects, who were HHV-8
and HLA A*0201-genotypic, or the 7 group C subjects, who were HHV-8+ but HLA A*0201 (median, 0; range, 0-20 spots/106 PBMCs). These results support that IFN-
production induced by gB492-500 was HHV-8-immune specific
and HLA A*0201-restricted.
Next, we determined that both PBMCs and CD8+ T cells, but
not CD4+ T cells, produced IFN-
We examined whether presence of HHV-8 DNA in the blood was associated with the ability of CD8+ T cells to respond to HHV-8 peptides. We tested PBMCs from a subset of 7 of the 14 group A persons for HHV-8 DNA, that is, subjects A25 through A30 and A34. We found that PBMCs from only 2 of these 7 subjects (A25 and A30) were positive for HHV-8 DNA. Thus, there was no association between HHV-8 DNAemia and ability of the CD8+ T cells to respond to these 25 HHV-8 peptides. Determination of the minimal length of the gB epitope Because the gB492-500 peptide was derived from a model of predicted binding of a 9-mer to an HLA motif, we next defined the true minimal epitope of this gB peptide by binding to HLA A*0201 and T-cell reactivity using six 9-mer peptides and two 10-mer peptides derived from overlapping sequences (Table 2). None of these overlapping peptides bound to the T2 cells (Figure 3). Nevertheless, we tested all 8 of these peptides for IFN- and CTL
reactivity in the event that these assays were more sensitive measures
of a peptide's T-cell stimulatory capacity. Our results show that greater IFN- production and CTL lysis was induced by the predicted, 9-mer gB492-500 peptide than by the overlapping 9-mer and
10-mer peptides in PBMCs of HHV-8+, HLA A*0201 individuals
from group A (Figure 4A,B). Some of the peptides (eg, gB491-500) induced low CTL and IFN-
reactivity even though they did not bind to HLA A*0201 in the T2 cell
assay (Figure 3).
We next established CD8+ T-cell lines to test T-cell
reactivity to these overlapping HHV-8 peptides. Two cell lines
generated against gB492-500 had higher IFN- T-cell response to gB492-500 in HLA A*0201 HHV-8 seropositive persons using gB492-500 peptide-loaded DCs We and others have previously shown that human blood monocyte-derived DCs loaded with HIV-1,35 EBV,36,37 and human cytomegalovirus (CMV)38,39 peptides are potent inducers of antiviral CD8+ T-cell responses in PBMCs of individuals seropositive for these viruses. Therefore, we determined if autologous DCs loaded with gB492-500 peptide were able to induce peptide-specific T-cell reactivity as compared to the conventional method of stimulation with peptide alone. Peptide-loaded and nonloaded, CD40L-matured DCs were added to autologous PBMCs derived from 5 group A and 4 group B subjects for 1 day (S0) and 7 days (S1), and tested in a single-cell IFN- assay. As shown in Figure 5A,
PBMCs from all 5 HHV-8 seropositive, group A subjects responded to
gB492-500 when the peptide was presented by autologous DCs
and cocultured for 1 week (S1; P = .004, 2-tailed
t test, as compared to group B subjects). Coculture of the PBMCs
overnight with peptide alone, or with peptide-loaded, autologous DCs
(S0), was insufficient to induce anti-gB492-500 T-cell
responses except in PBMCs from subject A52. There was little or no
response of PBMCs from the 4 HHV-8 group B seronegative donors after
stimulation with peptide-loaded, autologous DCs either overnight or for
7 days (Figure 5B). PBMCs from all 9 subjects responded strongly to
stimulation with PMA-iono (median, 2225; range, 1150-5520 spots/106 cells). None of these PBMCs were positive for
HHV-8 DNA.
IFN- in response to the gB492-500 peptide alone, but did produce IFN- in response to 1 week of stimulation with gB492-500 peptide-labeled DCs (Figure 5).
Further assessment using the DC-peptide system was therefore done to
determine the persistence of anti-gB492-500 T-cell
responses over time in 2 HHV-8+, HLA A*0201 individuals
(A27 and A29). Fresh DCs were obtained from the blood of each subject
and used to stimulate PBMCs that had been cryopreserved at
approximately 6-month intervals over more than 2 years. The data show
that IFN- production to gB492-500-loaded, autologous DCs
in the 7-day assay was consistent over time (Figure 6A,B). Moreover, the numbers of
IFN--producing cells were higher at all time points compared to
stimulation of PBMCs directly with the free peptide, supporting the
higher efficiency of T-cell activation by the antigen-expressing DCs.
The subjects' PBMCs also had consistently high responses to PMA-iono
over this time period (median, 2220; range, 1900-2500 spots/106 cells).
We examined whether HHV-8 DNA in blood was related to persistence of anti-HHV-8 CD8+ T-cell responses. The results indicate that all sequential PBMC samples from subjects A27 and A29 were negative for HHV-8 DNA (data not shown).
We provide evidence for an HLA A*0201-restricted,
CD8+ T-cell epitope specific for an HHV-8 lytic cycle
protein, gB. Twenty-five 9-mer peptides derived from 5 lytic cycle
proteins were selected for screening based on an HLA motif,
peptide-binding prediction model.27,28 One of the 25 HHV-8
peptides, gB492-500 (LMWYELSKI), was clearly identified as
immunogenic based on positive T-cell reactivity in HHV-8+,
HLA A*0201 individuals. This response was shown to be HHV-8-immune specific and HLA A*0201-restricted in that there was no IFN- Of importance is that our data support gB492-500 being a
major immunogenic HLA A*0201 epitope for HHV-8-specific,
CD8+ T cells. This was demonstrated by induction of high
numbers of IFN- We postulated that presence of latent HHV-8 infection in the blood may be related to the ability of the host's CD8+ T cells to respond to HHV-8 peptides. We did not find, however, a relationship of HHV-8 DNA in PBMCs with reactivity of T cells to these HHV-8 peptides. Indeed, most of our HHV-8-seropositive, healthy subjects were negative for HHV-8 DNA in blood by our PCR assay that can detect as few as 10 copies of HHV-8 DNA per 1 × 106 cells. This suggests that an infrequent, low level of latent HHV-8 infection occurs in the blood of healthy seropositive individuals, similar to other reports.46 This is in contrast to frequent detection of EBV47 and CMV48 DNA in the blood of asymptomatic carriers by highly sensitive PCR assays. Such a model of HHV-8 latency would predict a lower frequency and level of viral reactivation. This could account at least in part for the lower anti-HHV-8 T-cell responses detected in healthy, HHV-8-seropositive individuals,2-4 as compared to T-cell responses to EBV and CMV peptides in healthy, EBV- and CMV-seropositive persons.36-39 Although the precise function of HHV-8 gB is unknown, a recent report shows that it is part of the virion and infected cell membrane,49 in contrast to gB of EBV and MHV-68.50,51 Indeed, gB may be important for HHV-8 infectivity and pathogenesis in that it binds to cell surface heparan sulfate molecules.49 Regardless of its function in HHV-8, it is likely that the immune response to gB is related to the level of HHV-8 lytic replication. Several studies have shown an association of DNA positivity and higher levels of HHV-8 DNA in blood and tissues with development of KS and other HHV-8-related diseases.52-56 We hypothesize that such viral replication may boost CD8+ T-cell responses against HHV-8 lytic cycle proteins. This T-cell reactivity in turn acts to quell viral replication and prevent development of disease. Similarly, CD8+ T cells specific for peptides of EBV lytic cycle antigens have been observed during primary and latent EBV infection,7,16,17,57 suggesting that this immune response is involved in containment of EBV replication and related diseases. CTL reactivity to lytic cycle proteins is also postulated to be important in control of viral replication during acute, primary infection with MHV-68.10 Our study demonstrates that there is a novel HLA A*0201-restricted, immunogenic CD8+ T-cell epitope for HHV-8 lytic cycle protein gB. The identification of such CD8+ T-cell epitopes for HHV-8 proteins will be useful to our understanding of the role of HLA class I-restricted immunity in development of KS and other HHV-8-related diseases. We2,3 and others4 have previously shown that HHV-8 lytic cycle proteins gB, gH, MCP, MiCP, IE, and K8.1 elicit CD8+ T-cell responses during primary and persistent HHV-8 infections. A vaccine approach based on such T-cell-specific targets of the virus could potentially prevent HHV-8 infection, reactivation, and the development of associated diseases.
We thank Dr Xiao-Qing Zhao, Aki Hoji, Christine Kalinyak, Luann Borowski and Susan McQuiston for their research suggestions and technical assistance, and Judy Malenka for secretarial assistance. We also thank Bill Buchanan for clinical assistance and the Pitt Men's Study MACS staff and volunteers for their dedication and support. This work was done as part of the requirements for the doctorate degree by Q.J.W. in the Department of Infectious Diseases and Microbiology of the University of Pittsburgh Graduate School of Public Health.
Submitted June 29, 2001; accepted January 3, 2002.
Supported in part by National Institutes of Health grants P30 CA47904, R01 CA82053, R01 CA75957, and R03 CA81600.
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: Charles R. Rinaldo, Jr, Department of Infectious Diseases and Microbiology, Graduate School of Public Health, University of Pittsburgh, A427 Crabtree Hall, 130 DeSoto St, Pittsburgh, PA 15261; e-mail: rinaldo+{at}pitt.edu.
1. Sarid R, Olsen SJ, Moore PS. Kaposi's sarcoma-associated herpesvirus: epidemiology, virology, and molecular biology. Adv Virus Res. 1999;52:139-232[Medline] [Order article via Infotrieve]. 2. Wang QJ, Jenkins FJ, Jacobson LP, et al. CD8+ cytotoxic T lymphocyte responses to lytic proteins of human herpesvirus 8 in HIV-1 infected and uninfected individuals. J Infect Dis. 2000;182:928-932[CrossRef][Medline] [Order article via Infotrieve].
3.
Wang QJ, Jenkins FJ, Jacobson LP, et al.
Primary human herpesvirus 8 infection generates broadly specific CD8+ T cell response to viral lytic proteins.
Blood.
2001;97:2366-2373
4.
Osman M, Kubo T, Gill J, et al.
Identification of human herpesvirus 8-specific cytotoxic T-cell responses.
J Virol.
1999;73:6136-6140 5. Strickler HD, Goedert JJ, Bethke FR, et al. Human herpesvirus 8 cellular immune responses in homosexual men. J Infect Dis. 1999;180:1682-1685[CrossRef][Medline] [Order article via Infotrieve].
6.
Steven NM, Annels NE, Kumar A, Leese AM, Kurilla MG, Rickinson AB.
Immediate early and early lytic cycle proteins are frequent targets of the Epstein-Barr virus-induced cytotoxic T cell response.
J Exp Med.
1997;185:1605-1617 7. Khanna R, Sherritt M, Burrows SR. EBV structural antigens, gp350 and gp85, as targets for ex vivo virus-specific CTL during acute infectious mononucleosis: potential use of gp350/gp85 CTL epitopes for vaccine design. J Immunol. 1999;3063-3069.
8.
Pepperl S, Benninger-Doring G, Modrow S, Wolf H, Jilg W.
Immediate-early transactivator Rta of Epstein-Barr virus (EBV) shows multiple epitopes recognized by EBV-specific cytotoxic T lymphocytes.
J Virol.
1998;72:8644-8649 9. Bogedain C, Wolf C, Modrow S, Stuber G, Jilg W. Specific cytotoxic T lymphocytes recognize the immediate-early transactivator Zta of Epstein-Barr virus. J Virol. 1996;69:4872-4879[Abstract].
10.
Liu L, Flano E, Usherwood EJ, Surman S, Blackman MA, Woodland DL.
Lytic cycle T cell epitopes are expressed in two distinct phases during MHV-68 infection.
J Immunol.
1999;163:868-874 11. Rickinson AB, Moss DJ. Human cytotoxic T lymphocyte responses to Epstein- Barr virus infection. Annu Rev Immunol. 1997;15:405-431[CrossRef][Medline] [Order article via Infotrieve].
12.
Steven NM, Leese AM, Annels NE, Lee SP, Rickinson AB.
Epitope focusing in the primary cytotoxic T cell response to Epstein-Barr virus and its relationship to T cell memory.
J Exp Med.
1996;184:1801-1813 13. Yewdell JW, Norbury CC, Bennink JR. Mechanisms of exogenous antigen presentation by MHC class I molecules in vitro and in vivo: implications for generating CD8+ T cell responses to infectious agents, tumors, transplants, and vaccines. Adv Immunol. 1999;73:1-77[Medline] [Order article via Infotrieve]. 14. Benninger-Doring G, Pepperl S, Deml L, Modrow S, Wolf H, Jilg W. Frequency of CD8+ T lymphocytes specific for lytic and latent antigens of Epstein-Barr virus in healthy virus carriers. Virology. 1999;264:289-297[CrossRef][Medline] [Order article via Infotrieve].
15.
Yang J, Lemas VM, Flinn IW, Krone C, Ambinder RF.
Application of the ELISPOT assay to the characterization of CD8+ responses to Epstein-Barr virus antigens.
Blood.
2000;95:241-248 16. Callan MF, Steven N, Krausa P, et al. Large clonal expansions of CD8+ T cells in acute infectious mononucleosis. Nat Med. 1996;2:906-911[CrossRef][Medline] [Order article via Infotrieve].
17.
Tan LC, Gudgeon N, Annels NE, et al.
A re-evaluation of the frequency of CD8+ T cells specific for EBV in healthy virus carriers.
J Immunol.
1999;162:1827-1835
18.
Ehtisham S, Sunil-Chandra NP, Nash AA.
Pathogenesis of murine gammaherpesvirus infection in mice deficient in CD4 and CD8 T cells.
J Virol.
1993;67:5247-5252 19. Stevenson PG, Belz GT, Altman JD, Doherty PC. Changing patterns of dominance in the CD8+ T cell response during acute and persistent murine gamma- herpesvirus infection. Eur J Immunol. 1999;29:1059-1067[CrossRef][Medline] [Order article via Infotrieve].
20.
Liu L, Usherwood EJ, Blackman MA, Woodland DL.
T-cell vaccination alters the course of murine herpesvirus 68 infection and the establishment of viral latency in mice.
J Virol.
1999;73:9849-9857 21. Imanishi T, Akaz T, Kimura A, Tokunaga K, Gojobori T. Allele and haplotype frequencies for HLA and complement loci in various ethnic groups. In: Tsuji K,Aizawa M,Sasazuki T, eds. HLA. Vol 1. Oxford, England: Oxford University Press; 1991:1065. 22. Kaslow RA, Ostrow DG, Detels R, Phair JP, Polk BF, Rinaldo CR Jr. The Multicenter AIDS Cohort Study: rationale, organization, and selected characteristics of the participants. Am J Epidemiol. 1987;126:310-318.
23.
Rinaldo CR Jr, Huang XL, Fan Z, et al.
Anti-human immunodeficiency virus type 1 (HIV-1) CD8+ T-lymphocyte reactivity during combination antiretroviral therapy in HIV-1-infected patients with advanced immunodeficiency.
J Virol.
2000;74:4127-4138
24.
Chang Y, Cesarman E, Pessin MS, et al.
Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma.
Science.
1994;266:1865-1869
25.
Cesarman E, Chang Y, Moore PS, Said JW, Knowles DM.
Kaposi's sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas.
N Engl J Med.
1995;332:1186-1191
26.
Russo JJ, Bohenzky RA, Chien MC, et al.
Nucleotide sequence of the Kaposi's sarcoma-associated herpesvirus (HHV-8).
Proc Natl Acad Sci U S A.
1996;93:14862-14867 27. Parker KC, Bednarek MA, Hull K, et al. Sequence motifs important for peptide binding to the human MHC class I molecule, HLA-A2. J Immunol. 1992;149:3580-3587[Abstract]. 28. Rammensee HG, Friede T, Stevanoviic S. MHC ligands and peptide motifs: first listing. Immunogenetics. 1995;41:178-228[Medline] [Order article via Infotrieve]. 29. Zeh HJ, Leder GH, Lotze MT, et al. Flow-cytometric determination of peptide-class I complex formation: identification of p53 peptides that bind to HLA-A2. Human Immunol. 1994;39:79-86[CrossRef][Medline] [Order article via Infotrieve]. 30. Forero L, Zwirner NW, Fink CW, Fernandez-Vina MA, Stastny P. Juvenile arthritis, HLA-A2 and binding of DEK oncogene-peptides. Human Immunol. 1998;59:443-450[CrossRef][Medline] [Order article via Infotrieve]. 31. Salter RD, Cresswell P. Impaired assembly and transport of HLA-A and -B antigens in a mutant TxB cell hybrid. EMBO J. 1986;5:943-949[Medline] [Order article via Infotrieve]. 32. Buseyne F, Blanche S, Schmitt D, Griscelli C, Riviere Y. Detection of HIV-specific cell-mediated cytotoxicity in the peripheral blood from infected children. J Immunol. 1993;150:3569-3581[Abstract]. 33. Bednarek MA, Sauma SY, Gammon MC, et al. The minimum peptide epitope from the influenza virus matrix protein. Extra and intracellular loading of HLA-A2. J Immunol. 1991;147:4047-4053[Abstract].
34.
Fan Z, Huang XL, Borowski L, Mellors JW, Rinaldo CR Jr.
Restoration of anti-human immunodeficiency virus type 1 (HIV-1) responses in CD8+ T cells from late-stage patients on prolonged antiretroviral therapy by stimulation in vitro with HIV-1 protein-loaded dendritic cells.
J Virol.
2001;75:4413-4419 35. Fan Z, Huang XL, Zheng L, et al. Cultured blood dendritic cells retain HIV-1 antigen-presenting capacity for memory CTL during progressive HIV-1 infection. J Immunol. 1997;159:4973-4982[Abstract].
36.
Subklewe M, Paludan C, Tsang ML, Mahnke K, Steinman RM, Munz C.
Dendritic cells cross-present latency gene products from Epstein-Barr virus-transformed B cells and expand tumor-reactive CD8(+) killer T cells.
J Exp Med.
2001;193:405-411
37.
Redchenko IV, Rickinson AB.
Accessing Epstein-Barr virus-specific T-cell memory with peptide-loaded dendritic cells.
J Virol.
1999;73:334-342 38. Kleihauer A, Grigoleit U, Hebart H, et al. Ex vivo generation of human cytomegalovirus-specific cytotoxic T cells by peptide-pulsed dendritic cells. Br J Haematol. 2001;113:231-239[CrossRef][Medline] [Order article via Infotrieve]. 39. Vannucchi AM, Glinz S, Bosi A, Caporale R, Rossi-Ferrini P. Selective ex vivo expansion of cytomegalovirus-specific CD4+ and CD8+ T lymphocytes using dendritic cells pulsed with a human leucocyte antigen A*0201-restricted peptide. Br J Haematol. 2001;113:479-482[CrossRef][Medline] [Order article via Infotrieve].
40.
Larsson M, Messmer D, Somersan S, et al.
Requirement of mature dendritic cells for efficient activation of influenza A-specific memory CD8+ T cells.
J Immunol.
2000;165:1182-1190 41. Jin X, Roberts CG, Nixon DF, et al. Identification of subdominant cytotoxic T lymphocyte epitopes encoded by autologous HIV type 1 sequences, using dendritic cell stimulation and computer-driven algorithm. AIDS Res Hum Retroviruses. 2000;16:67-76[CrossRef][Medline] [Order article via Infotrieve]. 42. Banchereau J, Briere F, Caux C, et al. Immunobiology of dendritic cells. Annu Rev Immunol. 2000;18:767-811[CrossRef][Medline] [Order article via Infotrieve].
43.
Mosca PJ, Hobeika AC, Clay TM, et al.
A subset of human monocyte-derived dendritic cells expresses high levels of interleukin-12 in response to combined CD40 ligand and interferon-gamma treatment.
Blood.
2000;96:3499-3504 44. Kuniyoshi JS, Kuniyoshi CJ, Lim AM, et al. Dendritic cell secretion of IL-15 is induced by recombinant huCD40LT and augments the stimulation of antigen-specific cytolytic T cells. Cell Immunol. 1999;193:48-58[CrossRef][Medline] [Order article via Infotrieve]. 45. Brander C, O'Connor P, Suscovich T, et al. Definition of an optimal cytotoxic T lymphocyte epitope in the latently expressed Kaposi's sarcoma-associated herpesvirus kaposin protein. J Infect Dis. 2001;184:119-126[CrossRef][Medline] [Order article via Infotrieve].
46.
Decker LL, Shankar P, Khan G, et al.
The Kaposi's sarcoma-associated herpesvirus (KSHV) is present as an intact latent genome in KS tissue but replicates in the peripheral blood mononuclear cells of KS patients.
J Exp Med.
1996;184:283-288 47. Khan G, Miyashita EM, Yang B, Babcock GJ, Thorley-Lawson DA. Is EBV persistence in vivo a model for B cell homeostasis? Immunity. 1996;5:173-179[CrossRef][Medline] [Order article via Infotrieve]. 48. Larsson S, Soderberg-Naucler C, Wang FZ, Moller E. Cytomegalovirus DNA can be detected in peripheral blood mononuclear cells from all seropositive and most seronegative healthy blood donors over time. Transfusion. 1998;38:271-278[CrossRef][Medline] [Order article via Infotrieve]. 49. Akula SM, Pramod NP, Wang F-Z, Chandran B. Human herpesvirus 8 envelope-associated glycoprotein B interacts with heparan sulfate-like moieties. Virology. 2001;284:235-249[CrossRef][Medline] [Order article via Infotrieve].
50.
Gong M, Kieff E.
Intracellular trafficking of two major Epstein-Barr virus glycoproteins, gp350/220 and gp110.
J Virol.
1990;64:1507-1516
51.
Stewart JP, Janjua NJ, Sunil-Chandra NP, Nash AA, Arrand JR.
Characterization of murine gammaherpesvirus 68 glycoprotein B (gB) homolog: similarity to Epstein-Barr virus gB (gp110).
J Virol.
1994;68:6496-6504 52. Moore PS, Kingsley LA, Holmberg SD, et al. Kaposi's sarcoma-associated herpesvirus infection prior to onset of Kaposi's sarcoma. AIDS. 1996;10:175-180[Medline] [Order article via Infotrieve]. 53. Boivin G, Gaudreau A, Routy JP. Evaluation of the human herpesvirus 8 DNA load in blood and Kaposi's sarcoma skin lesions from AIDS patients on highly active antiretroviral therapy. AIDS. 2000;14:1907-1910[CrossRef][Medline] [Order article via Infotrieve]. 54. Campbell TB, Borok M, Gwanzura L, et al. Relationship of human herpesvirus 8 peripheral blood virus load and Kaposi's sarcoma clinical stage. AIDS. 2000;14:2109-2116[CrossRef][Medline] [Order article via Infotrieve]. 55. Pozo F, Tenorio A, de la Mata M, de Ory F, Torre-Cisneros J. Persistent human herpesvirus 8 viremia before Kaposi's sarcoma development in a liver transplant recipient. Transplantation. 2000;70:395-397[CrossRef][Medline] [Order article via Infotrieve].
56.
Oksenhendler E, Carcelain G, Aoki Y, et al.
High levels of human herpesvirus 8 viral load, human interleukin-6, interleukin-10, and C reactive protein correlate with exacerbation of multicentric Castleman disease in HIV-infected patients.
Blood.
2000;96:2069-2073
57.
Toussirot E, Wendling D, Tiberghien P, Luka J, Roudier J.
Decreased T cell precursor frequencies to Epstein-Barr virus glycoprotein gp110 in peripheral blood correlate with disease activity and severity in patients with rheumatoid arthritis.
Ann Rheum Dis.
2000;59:533-538
© 2002 by The American Society of Hematology.
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  G. Rappocciolo, H. R. Hensler, M. Jais, T. A. Reinhart, A. Pegu, F. J. Jenkins, and C. R. Rinaldo Human Herpesvirus 8 Infects and Replicates in Primary Cultures of Activated B Lymphocytes through DC-SIGN J. Virol., May 15, 2008; 82(10): 4793 - 4806. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Barcy, S. C. De Rosa, J. Vieira, K. Diem, M. Ikoma, C. Casper, and L. Corey {gamma}{delta}+ T Cells Involvement in Viral Immune Control of Chronic Human Herpesvirus 8 Infection J. Immunol., March 1, 2008; 180(5): 3417 - 3425. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Guihot, E. Oksenhendler, L. Galicier, A.-G. Marcelin, L. Papagno, A.-S. Bedin, F. Agbalika, N. Dupin, J. Cadranel, B. Autran, et al. Multicentric Castleman disease is associated with polyfunctional effector memory HHV-8-specific CD8+ T cells Blood, February 1, 2008; 111(3): 1387 - 1395. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Stebbing, A. Sanitt, A. Teague, T. Powles, M. Nelson, B. Gazzard, and M. Bower Prognostic Significance of Immune Subset Measurement in Individuals With AIDS-Associated Kaposi's Sarcoma J. Clin. Oncol., June 1, 2007; 25(16): 2230 - 2235. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Lambert, M. Gannage, A. Karras, M. Abel, C. Legendre, D. Kerob, F. Agbalika, P.-M. Girard, C. Lebbe, and S. Caillat-Zucman Differences in the frequency and function of HHV8-specific CD8 T cells between asymptomatic HHV8 infection and Kaposi sarcoma Blood, December 1, 2006; 108(12): 3871 - 3880. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Rappocciolo, F. J. Jenkins, H. R. Hensler, P. Piazza, M. Jais, L. Borowski, S. C. Watkins, and C. R. Rinaldo Jr DC-SIGN Is a Receptor for Human Herpesvirus 8 on Dendritic Cells and Macrophages J. Immunol., February 1, 2006; 176(3): 1741 - 1749. [Abstract] [Full Text] [PDF]
|
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|
|
|
|
E. Ribechini, C. Fortini, M. Marastoni, S. Traniello, S. Spisani, P. Monini, and R. Gavioli Identification of CD8+ T Cell Epitopes within Lytic Antigens of Human Herpes Virus 8 J. Immunol., January 15, 2006; 176(2): 923 - 930. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X.-L. Huang, Z. Fan, B. A. Colleton, R. Buchli, H. Li, W. H. Hildebrand, and C. R. Rinaldo Jr. Processing and Presentation of Exogenous HLA Class I Peptides by Dendritic Cells from Human Immunodeficiency Virus Type 1-Infected Persons J. Virol., March 1, 2005; 79(5): 3052 - 3062. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Wang, H. Chen, X. Jiang, M. Zhang, T. Wan, N. Li, X. Zhou, Y. Wu, F. Yang, Y. Yu, et al. Identification of an HLA-A*0201-restricted CD8+ T-cell epitope SSp-1 of SARS-CoV spike protein Blood, July 1, 2004; 104(1): 200 - 206. [Abstract] [Full Text] [PDF]
|
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Y.-D. Wang, W.-Y. F. Sin, G.-B. Xu, H.-H. Yang, T.-y. Wong, X.-W. Pang, X.-Y. He, H.-G. Zhang, J. N. L. Ng, C.-S. S. Cheng, et al. T-Cell Epitopes in Severe Acute Respiratory Syndrome (SARS) Coronavirus Spike Protein Elicit a Specific T-Cell Immune Response in Patients Who Recover from SARS J. Virol., June 1, 2004; 78(11): 5612 - 5618. [Abstract] [Full Text] [PDF]
|
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J. Stebbing, B. Gazzard, S. Patterson, M. Bower, D. Perumal, M. Nelson, A. McMichael, G. Ogg, A. Epenetos, F. Gotch, et al. Antibody-targeted MHC complex-directed expansion of HIV-1- and KSHV-specific CD8+ lymphocytes: a new approach to therapeutic vaccination Blood, March 1, 2004; 103(5): 1791 - 1795. [Abstract] [Full Text] [PDF]
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 J. Stebbing, S. Portsmouth, and B. Gazzard How does HAART lead to the resolution of Kaposi's sarcoma? J. Antimicrob. Chemother., May 1, 2003; 51(5): 1095 - 1098. [Full Text] [PDF]
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J. Stebbing, D. Bourboulia, M. Johnson, S. Henderson, I. Williams, N. Wilder, M. Tyrer, M. Youle, N. Imami, T. Kobu, et al. Kaposi's Sarcoma-Associated Herpesvirus Cytotoxic T Lymphocytes Recognize and Target Darwinian Positively Selected Autologous K1 Epitopes J. Virol., April 1, 2003; 77(7): 4306 - 4314. [Abstract] [Full Text] [PDF]
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Abstract
Introduction
