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IMMUNOBIOLOGY
From the Departments of Hematology and Immunohematology
and Bloodbank, Leiden University Medical Center, Leiden, The
Netherlands.
Rejection of a graft after human leukocyte antigen
(HLA)-identical stem cell transplantation (SCT) can be caused by
recipient's immunocompetent T lymphocytes recognizing minor
histocompatibility antigens on donor stem cells. During rejection of a
male stem cell graft by a female recipient, 2 male (H-Y)-specific
cytotoxic T lymphocyte (CTL) clones were isolated from peripheral
blood. One CTL clone recognized an HLA-A2-restricted H-Y antigen,
encoded by the SMCY gene. Another CTL clone recognized an
HLA-B60-restricted H-Y antigen. In this study UTY was
identified as the gene coding for the HLA-B60-restricted H-Y antigen.
The UTY-derived H-Y antigen was characterized as a 10-amino
acid residue peptide, RESEEESVSL. Although the epitope differed by
3 amino acids from its X-homologue, UTX, only 2 polymorphisms were
essential for recognition by the CTL clone HLA-B60 HY. These
results illustrate that CTLs against several H-Y antigens derived from
different proteins can contribute simultaneously to graft rejection
after HLA-identical, sex-mismatched SCT. Moreover, RESEEESVSL-specific
T cells could be isolated from a female HLA-B60+ patient with
myelodysplastic syndrome who has been treated with multiple blood
transfusions, but not from control healthy HLA-B60+ female
donors. This may indicate that RESEEESVSL-reactive T cells are more
common in sensitized patients.
(Blood. 2000;96:3126-3132) Human leukocyte antigen (HLA)-identical
hematopoietic stem cell transplantation (SCT) can be complicated by
graft-versus-host disease (GVHD) or graft rejection. Graft rejection
can be caused by the recognition of minor histocompatibility antigens
(mHag) on donor stem cells by immunocompetent HLA-restricted T
lymphocytes of recipient origin.1-3 mHag are peptides
derived from polymorphic intracellular proteins that differ between
donor and recipient.4,5 The peptides are presented on the
cell membrane in association with major histocompatibility complex
molecules and are capable of eliciting specific T-cell
responses.2,4 Specific T-cell responses against
male-specific mHag may occur after sex-mismatched blood transfusions
and organ or stem cell transplantation.6-8 Clinical
studies indicated that female patients who underwent transplantation
with HLA-phenotypic identical male stem cells have a higher risk for
graft rejection than those who receive stem cells of female
origin.6,9 During rejection of male stem cell grafts,
H-Y-specific cytotoxic T lymphocyte (CTL) clones could be generated
from the peripheral blood of female patients.1,8,10 The
H-Y-specific CTL clones are used to identify the H-Y antigens and
their genes.
SMCY was the first gene identified that
encoded human H-Y antigens.11,12 HLA-A2- and
HLA-B7-restricted H-Y epitopes derived from the SMCY protein were
characterized through peptide elution of male cells, and microcapillary
liquid chromatography-electrospray ionization mass spectrometry
combined with T-cell epitope reconstitution. A molecular method can be
explored as well. The DNA sequences of 8 Y-specific genes are
known.13 Each gene has a ubiquitous tissue expression and
a homologue on the X chromosome encoding a similar, but nonidentical,
protein isoform. The amino acid sequence identity of the X-Y isoforms
varies from 85% to 97%. Because these ubiquitously expressed
Y-specific genes have sufficient polymorphisms to generate
male-specific peptides, they are potential candidates to encode H-Y antigens.
DFFRY was identified as a second gene encoding a human H-Y
antigen.14,15 The DFFRY-derived H-Y peptide was recognized
by an HLA-A1-restricted CTL clone, which was generated from a female patient with acute myeloid leukemia who rejected her
HLA-identical male stem cells.1 We identified the
HLA-A1-restricted H-Y epitope by DFFRY deletion mutants and
DFFRY minigenes encoding small peptides. Synthetic peptides
were used to identify this H-Y antigen recognized by the HLA-A1
HY-specific CTL clone. Recently, the UTY gene was identified as a gene coding for an H-Y epitope, which was recognized by
an HLA-B8-restricted CTL clone generated during GVHD.16
The first indication that multiple H-Y antigens may be involved in male
stem cell graft rejection in a female patient was the isolation of
HLA-A2- and HLA-B60-restricted cytotoxic and proliferative
H-Y-specific CTL clones from the patient, a woman with aplastic anemia
who rejected an HLA-genotypic identical male stem cell
graft.17,18 The H-Y epitope recognized by the
HLA-A2-restricted CTL clone was previously characterized as a peptide
derived from the SMCY gene.11 The H-Y-specific
HLA-B60-restricted CD8+ CTL clone showed no reactivity to
the SMCY protein, indicating that another Y-specific gene was also
involved in the male stem cell graft rejection.
In this study, we identified UTY as the gene encoding the
H-Y epitope recognized by the HLA-B60-restricted CTL clone. The H-Y
epitope was localized on the UTY gene using UTY
deletion mutants and UTY minigenes. Synthetic peptides
confirmed that the amino acid sequence of the UTY-derived H-Y epitope
was RESEEESVSL. Although the epitope contained 3 different amino acids
compared to its UTX-homologue protein, only 2 amino acids were shown to
be essential residues for recognition by the HLA-B60 H-Y CTL clone.
Furthermore, we demonstrated that RESEEESVSL-specific T cells could be
isolated from a female HLA-B60+ patient with myelodysplastic syndrome
(MDS) treated with multiple blood transfusions, and not from 2 healthy control HLA-B60+ female donors. This may reflect increased frequencies of RESEEESVSL-specific T cells in female HLA-B60+ patients sensitized by frequent stimulation with male cells.
CTL and cell lines
Cloning of Y-specific genes
Cloning of subgenic fragments or minigenes of the UTY gene Restriction analysis on the UTY gene revealed an HpaI site on position 3306, an NheI site on position 2458, and a PstI site on position 1569 of the UTY cDNA. NheI and PstI also have a recognition site in the multiple cloning site of the PCR3.1 plasmid, directly after the 5' and 3' ends of the UTY cDNA, respectively. Deletion mutants UTY/NheI and UTY/PstI were generated by incubating the plasmid PCR3.1 containing the UTY gene with the restriction enzymes NheI and PstI, deleting 2472-bp and 2493-bp long fragments at the 5' and 3' sites of the UTY gene, respectively. Deletion mutant UTY/HpaI was generated by digesting UTY with HpaI and the PCR3.1 plasmid with EcoRV directly after the 3' end of the UTY cDNA, deleting a 756-bp long fragment of the UTY gene. After digestion, the linear truncated constructs were visualized on an ethidium bromide-stained low-melt agarose gel. After isolating the constructs with agarose, the linear constructs were ligated with the rapid DNA ligation kit (Boehringer Mannheim GmbH) to form circular plasmid DNA consisting of vector PCR3.1 with the truncated UTY/HpaI, UTY/NheI, and UTY/PstI cDNAs.The oligonucleotides (5'
Transfection of HeLa cells and screening of transfectants Y-specific cDNA was transfected into HeLa cells by the DEAE-dextran-chloroquine method as described.21 One day before transfection, HeLa cells were seeded in 96-well flat-bottom microtiter plates at 15 000 cells/well in 100 µL complete culture medium. Before transfection, medium was discarded and replaced by 45 µL DEAE-dextran/DNA mixtures. These mixtures were prepared for duplicate transfections in 96-well V-bottom microtiter plates by sequentially adding: 98 µL RPMI-1640 supplemented with 10% decomplemented NuSerum IV (Collaborative Biomedical Products, Bedford, MA) containing 0.3 mg/mL DEAE-dextran (Sigma Chemical) and 100 µmol/L chloroquine, 1 µL of TE (10 mmol/L Tris, 1 mmol/L EDTA, pH 7.4) containing 200 ng Y-specific cDNA in plasmid PCR3.1 and 1 µL TE containing 200 ng plasmid PCR3.1-B60 (plasmid PCR3.1 containing the HLA-B60 gene) or without HLA-B60 construct as a control. The HeLa cells were incubated for 4 hours at 37°C, after which the DEAE-dextran/DNA was discarded and replaced by 80 µL phosphate-buffered saline (PBS) containing 10% dimethyl sulfoxide (DMSO). After 2 minutes at room temperature, PBS-DMSO was replaced by 200 µL complete culture medium. Transfected HeLa cells were incubated for 48 hours at 37°C. After removing the medium, 4000 HLA-B60 HY CTL were added in 200 µL RPMI-1640 containing 10% pooled human serum and 300 IU IL-2 per milliliter. After 24 hours, the tumor necrosis factor (TNF) content in 50 µL supernatant was determined by adding it to 50 µL complete culture medium containing 50 000 WEHI-164 clone 13 cells, sensitive to a cytolytic effect by TNF.22 After 18 hours, 10 µL cell proliferation reagent WST I (Boehringer Mannheim GmbH) was added, and the TNF concentration was calculated in comparison with recombinant TNF standards measured in the same assay.Peptide synthesis and peptide recognition assays Peptides were synthesized by solid-phase strategies on an automated multiple peptide synthesizer (SyroII; MultiSynTech, Witten, Germany) and characterized by mass spectrometry. The purity of the peptides was determined by analytical reversed-phase high-pressure liquid chromatography and it proved to be at least 80%. 51Cr-release assays were used to determine lysis of target cells. EBV-LCL target cells were labeled with 100 µCi of Na51CrO4. After 1 hour of incubation at 37°C, the cells were washed 3 times with RPMI supplemented with 2% FCS. Then 2000 Na51CrO4-labeled EBV-LCL cells/well were plated in a 96-well V-bottom microtiter plate in 100 µL RPMI-1640 + 10% pooled human serum containing various concentrations of peptide. After 1 hour of incubation at 37°C, 20 000 CTLs were added in 100 µL RPMI-1640 + 10% pooled human serum. 51Cr-release was measured after incubation at 37°C for 4 hours.Screening of UTY protein for HLA-B60-binding peptides The UTY protein was screened for HLA-B60-binding peptides by using the program HLA peptide-binding predictions (http://bimas.dcrt.nih.gov/molbio/hla_bind/). The scores are predictions of half-time of dissociation to HLA-B60 molecules for the indicated peptides.HLA-B60 binding assay Homozygous HLA-B60+ EBV cells were incubated at 4°C for 90 seconds in citric acid, pH 3.1, to elute peptides from the HLA class I complexes. After washing the cells twice with RPMI-1640 medium containing 2% FCS, the EBV cells were resuspended at a concentration of 400 000 cells/mL in Iscove minimum Dulbecco medium (IMDM) containing 2% FCS and 1.5 µg/mL 2-microglobulin. A total of 100 µL EBV cells were added to 25 µL 150 nmol/L of fluorescence-labeled HLA-B60-binding peptide KESTC(FL)HLVL in a 96-well, V-bottom
microtiter plate. For evaluation of maximal binding of the fluorescent
peptide to the EBV cells, 25 µL PBS was added to the wells. For
determination of peptide binding to HLA-B60 molecules, 25 µL PBS
containing peptide concentrations of 0.4 µg/mL to 50 µg/mL was
added to the wells. KESTLHLVL without fluorescence label was used as a
control. After incubation for 24 hours at 4°C, the cells were washed
twice with PBS, and the binding of the fluorescence-labeled
peptide to the EBV cells was determined by FACS analysis. The
concentration of a given peptide that induced a decrease of 50% of the
mean channel of fluorescence of maximal binding (IC50)
was calculated.
Isolation of IFN  -producing cells were isolated using the MACS IFN secretion
assay (Miltenyi Biotec, Auburn, CA) according to the manufacturer's
procedure. After isolation, IFN and IFN+ T cells were
stimulated with RESEEESVSL peptide-loaded autologous mononuclear bone
marrow cells in IMDM medium containing 10% human serum, 3 mmol/L
L-glutamine, and 120 U/mL recombinant IL-2. The T cells were
restimulated on days 7 and 14 with RESEEESVSL-loaded irradiated
autologous mononuclear bone marrow cells. On day 21, T-cell reactivity
against the RESEEESVSL epitope was determined in a 51Cr
release assay.
Cloning of Y-specific genes After isolation of RNA from male EBV-LCL cells, Y-gene-specific primers were used in a PCR reaction to amplify only the Y-specific genes and not their X-homologues. After cloning the PCR products in an expression vector, restriction and sequence analysis of the amplified Y-specific cDNAs confirmed their Y specificity (data not shown). As described previously, all Y-specific cDNAs were appropriately transcribed and translated as determined in an in vitro transcription/translation assay.14Cloning and identification of the gene encoding the HLA-B60-restricted H-Y T-cell epitope To determine whether any of the Y-specific genes encode the HLA-B60-restricted H-Y T-cell epitope, the cloned Y-cDNAs were cotransfected with HLA-B60 cDNA into HeLa cells. As shown in Figure 1, transfection of UTY cDNA resulted in a significant amount of TNF production by HLA-B60 HY CTL, whereas transfection of all other Y genes induced a TNF release of 1 pg/mL. Deletion mutants of the UTY sequence were obtained by digestion with restriction enzymes HpaI and PstI, resulting in a deletion at the 3' site of the UTY gene of 756 bp (UTY/HpaI) and 2493 bp (UTY/PstI), respectively (Figure 2). An additional deletion of 2472 bp at the 5' site of the UTY gene (UTY/NheI) was generated using the restriction enzyme NheI. As is demonstrated in Figure 3, no TNF production by the HLA-B60 HY CTL clone was measured when stimulated with HeLa transfected with UTY/NheI and HLA-B60. Transfection of UTY/PstI in HeLa resulted in TNF release by the HLA-B60 HY CTL, illustrating that the HLA-B60-restricted H-Y peptide was encoded by an amino acid sequence between positions 1 and 523 of the UTY protein. To confirm that the TNF release by the HLA-B60 HY CTL was HLA-B60 restricted, the UTY cDNAs were also transfected without HLA-B60 into HeLa cells. No significant TNF release by the HLA-B60 HY CTL was found when the UTY cDNAs were transfected in the absence of the HLA-B60 molecule (Figure 3).
Localization of the HLA-B60-restricted H-Y epitope in the UTY gene To identify the H-Y peptide coding sequence, the 1-523 amino acid (AA) region of the UTY protein was screened for HLA-B60-binding peptides that differ by at least one amino acid from its X homologue. Twelve peptides (9 or 10 residues) with scores varying from 17 600 to 704 000 were observed (Table 2). Three minigenes coding for 8 HLA-B60-binding peptides were constructed and cotransfected with HLA-B60 into HeLa cells (Figure 2). As is demonstrated in Figure 4, only minigene 79-120, coding for peptide MASRESEEESVSLTV, induced TNF production by HLA-B60 HY CTL, indicating that this minigene encoded the H-Y epitope recognized by the HLA-B60 HY CTL clone. Transfection of the minigene without cotransfection of the HLA-B60 molecule into HeLa cells did not result in TNF production.
Identification of the H-Y epitope recognized by CTL clone HLA-B60 HY Minigene 79-120, coding for peptide MASRESEEESVSLTV, coded for 3 HLA-B60-binding peptides (Table 2). The relevant peptides were synthesized and loaded at various concentrations on female HLA-B60+ EBV-LCL cells. Recognition of the peptide loaded EBV-LCL cells by the CTL clone HLA-B60 HY was determined in a 51Cr-release assay. As is shown in Figure 5A, the RESEEESVSL-loaded EBV-LCL cells were efficiently lysed by HLA-B60 HY CTL with half-maximal target cell lysis at a peptide concentration of 300 pg/mL, whereas neither EEESVSLTV nor the SEEESVSLTV peptide was recognized by the HLA-B60 HY CTL. To determine whether the 10-residue peptide RESEEESVSL was the minimal epitope recognized by HLA-B60 HY CTL, 2 peptides lacking the N- or C-terminal amino acid of RESEEESVSL were synthesized and tested for recognition by HLA-B60 HY CTL in a 51Cr release assay. As demonstrated in Figure 5B, EBV-LCL cells loaded with the 10-residue peptide RESEEESVSL were most efficiently lysed by the HLA-B60 HY CTL.
Two polymorphisms in H-Y epitope are essential for T-cell recognition The H-Y epitope RESEEESVSL differed by 3 amino acids from its X homologue GESEEASPSL, which was not recognized by the HLA-B60 HY CTL clone (Figure 6A). To determine whether all 3 polymorphisms were equally important for recognition by the HLA-B60 HY CTL, all 3 peptides each containing one UTX-homologue amino
acid on position 1, 6, or 8were synthesized. As shown in Figure 6A,
EBV-LCL cells loaded with peptide GESEEESVSL were well recognized by
HLA-B60 HY CTL. In contrast, an X-homologue amino acid on position 6 or 8 of the H-Y epitope resulted in a reduced lysis of EBV-LCL cells by
the HLA-B60 HY CTL, suggesting that these polymorphisms were essential
for T-cell recognition. The peptides GESEEASVSL and GESEEESPSL, each
containing 2 X-homologue amino acids, were not recognized by HLA-B60 HY
CTL (Figure 6B). To determine whether the mutated peptides bind equally
well to HLA-B60 molecules as the original RESEEESVSL peptide, an assay
based on competition for binding to HLA-B60 between a peptide of
interest and a fluorescence-labeled standard peptide was used. Similar
to the standard peptide without fluorescence label, 0.8 µg/mL
RESEEESVSL competitor peptide induced 50% inhibition of
fluorescence-labeled peptide binding (IC50), suggesting a
strong binding of RESEEESVSL to HLA-B60 molecules. A control irrelevant
peptide did not compete for binding of the fluorescence-labeled
standard peptide to the HLA-B60 molecule. All mutated variants of
RESEEESVSL induced IC50 at concentrations varying from 0.6 to 0.8 µg/mL, indicating that the altered recognition of the mutated
peptides by HLA-B60 HY CTL was not a consequence of altered binding of
the peptides to HLA-B60 molecules.
Isolation of HLA-B60 HY CTL from a female patient with MDS To determine whether HLA-B60 HY CTL were present in female patients, PBMC of 2 healthy donors and a patient with MDS who was sensitized by multiple blood transfusions were stimulated with the RESEEESVSL epitope. After stimulation, activated T cells (IFN+) were
separated from nonreactive T cells (IFN) based on their IFN
production. The frequency of IFN-producing T cells in PBMC of donors
and patient was below the detection level, as measured by flow
cytometry (less than 0.05%; data not shown). After 20 days of culture,
FACS analysis showed that 20% to 30% of the T cells in all T-cell
lines were CD3+ and CD8+ (data not shown).
Reactivity of the T-cell lines against the RESEEESVSL epitope was
tested in a 51Cr-release assay. The IFN T-cell line
generated from PBMC of the patient with MDS lysed RESEEESVSL-loaded
autologous or allogeneic HLA-B60 female EBV cells, whereas no
reactivity could be observed against the control peptide (Figure
7). Moreover, HLA-B60+ EBV cells from a
male were also efficiently lysed by the T-cell line. The IFN
T-cell line derived from the same patient showed no reactivity against
the RESEEESVSL epitope (Figure 7). The IFN+ and IFN T-cell
lines derived from the 2 donors exhibited no reactivity against the
RESEEESVSL epitope (data not shown).
The involvement of male-specific mHag antigens in graft rejection was indicated by the observation that female patients who underwent transplantation with HLA-phenotypic identical male stem cells had a higher risk for graft rejection than those who underwent transplantation using stem cell grafts of female origin.6,9 T-cell clones recognizing male cells could be generated during stem cell graft rejection after sex-mismatched transplantation. H-Y-specific T cells can be used to identify the H-Y antigens and their genes.1,17 Two human H-Y epitopes have been identified. The H-Y epitopes, recognized by HLA-A2- and HLA-B7-restricted CTL clones, were shown to be encoded by the ubiquitously expressed SMCY gene.11,12 Recently, we identified DFFRY as a second male-specific protein capable of inducing an HLA-A1-restricted CTL response during stem cell graft rejection in humans.14 In this study we characterized UTY as the gene coding for the H-Y epitope recognized by an HLA-B60-restricted CTL clone. The CTL clone was generated from the same female patient, during rejection of the HLA-identical male stem cell graft, as the HLA-A2-restricted H-Y-specific CTL clone recognizing the SMCY-derived epitope. This illustrates that the immunogenic peptides from both the SMCY and the UTY gene were involved in the male stem cell graft rejection in one female patient. UTY maps to proximal Yq11 within the H-Y antigen-controlling locus HY.13 The complete coding region of UTY shows 86% identity at the amino acid level to the X-homologue UTX. The UTY gene is widely expressed and encodes a tetratricopeptide repeat protein.23 Tetratricopeptide motifs are found in a variety of functionally distinct proteins and are believed to mediate protein-protein interaction. Differential splicing of the UTY gene may generate 3 UTY isoforms differing at their C-termini. The N-terminus of the UTY isoforms is conserved. Because the HLA-B60-restricted H-Y epitope is located at the N-terminus, the 3 UTY isoforms encode the same H-Y epitope. Although UTY is ubiquitously expressed, the expression level in different tissues may be variable. An HLA-B8-restricted H-Y-specific CTL clone recognizing a UTY-encoding epitope lysed male hematopoietic cells efficiently, whereas no or limited reactivity could be detected against HLA-B8+ male fibroblasts.16,24 The reactivity of the HLA-B60-restricted H-Y-specific CTL clone against different cells and tissues has to be further evaluated. Although the UTY epitope contained 3 different amino acids compared with its UTX homologue, only 2 polymorphisms were essential residues for recognition by the HLA-B60 HY CTL. Substitution of the N-terminal amino acid of the H-Y epitope by its X-homologue amino acid did not affect recognition by the H-Y-specific CTL. Substitution of the other 2 polymorphisms resulted in a decreased CTL recognition of the mutated H-Y peptides. This was not due to a decreased binding of the mutated peptides to HLA-B60, as was demonstrated in HLA-B60 binding studies. The single mutated H-Y epitopes were only recognized when high peptide concentrations were used. Additional substitution of the N-terminal amino acid in the latter peptides by its X-homologue completely abolished T-cell recognition. This indicates that the N-terminal amino acid does contribute to the 3-dimensional structure of the HLA-peptide complex recognized by the specific CTL clone. This contribution was only detectable using suboptimal single mutated epitopes. The identification of UTY as a third gene encoding an H-Y epitope demonstrates that multiple male antigens derived from different Y-specific genes are involved in male stem cell graft rejection. The role of these different H-Y antigens in stem cell graft rejection must be further evaluated. H-Y-specific T cells present in female patients during male graft rejection can be monitored by using HLA/HY peptide tetramerics.25 Tetrameric complexes of HLA-A2 or HLA-B7 with SMCY-derived H-Y epitopes were used to monitor H-Y-specific T cells during GVHD of male patients treated with female stem cells. A significant increase of H-Y-specific CTL during acute and chronic GVHD could be visualized.25 Identification of H-Y-specific T cells in female patients before SCT may be useful in the selection of donor-recipient combinations. This would especially be relevant in female patients who may be sensitized against male antigens by multiple blood transfusions before SCT.1,26 We demonstrated that RESEEESVSL-specific T cells could be isolated from an HLA-B60+ female patient with MDS who received multiple blood transfusions, and not from 2 healthy control HLA-B60+ female donors. Because the frequency of RESEEESVSL-specific T cells in the patient was very low (less than 0.05%), direct analysis of the frequencies of H-Y-specific T cells by flow cytometry was not possible. However, we were able to isolate and culture RESEEESVSL-specific T cells within 3 weeks from the patients PBMC and not from the control donors, indicating an increased frequency of RESEEESVSL-specific T cells in the patient. Transplantation of HLA-identical male stem cells in such patients may lead to activation of these RESEEESVSL-specific T cells, which may induce rejection of a male stem cell graft.
We thank Bregje Mommaas and Jan Wouter Drijfhout (Immunohematology and Bloodbank, Leiden University Medical Center, Leiden, The Netherlands) for providing the fluorescence-labeled HLA-B60-binding peptide.
Submitted December 30, 1999; accepted July 10, 2000.
Supported by the Dutch Cancer Society (grant RUL 99-2028) and the J.A. Cohen Institute for Radiopathology and Radiation Protection.
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: Mario H. J. Vogt, Department of Hematology, Leiden University Medical Center, C2-R, PO Box 9600, 2300 RC Leiden, The Netherlands; e-mail: m.h.j.vogt{at}lumc.nl.
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© 2000 by The American Society of Hematology.
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  K. V. Rosinski, N. Fujii, J. K. Mito, K. K. W. Koo, S. M. Xuereb, O. Sala-Torra, J. S. Gibbs, J. P. Radich, Y. Akatsuka, B. J. Van den Eynde, et al. DDX3Y encodes a class I MHC-restricted H-Y antigen that is expressed in leukemic stem cells Blood, May 1, 2008; 111(9): 4817 - 4826. [Abstract] [Full Text] [PDF]
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K. Rezvani, A. S. M. Yong, B. N. Savani, S. Mielke, K. Keyvanfar, E. Gostick, D. A. Price, D. C. Douek, and A. J. Barrett Graft-versus-leukemia effects associated with detectable Wilms tumor-1 specific T lymphocytes after allogeneic stem-cell transplantation for acute lymphoblastic leukemia Blood, September 15, 2007; 110(6): 1924 - 1932. [Abstract] [Full Text] [PDF]
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J. C. Zimring, G. A. Hair, S. S. Deshpande, and J. T. Horan Immunization to minor histocompatibility antigens on transfused RBCs through crosspriming into recipient MHC class I pathways Blood, January 1, 2006; 107(1): 187 - 189. [Abstract] [Full Text] [PDF]
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 M. Millrain, D. Scott, C. Addey, H. Dewchand, P. Ellis, I. Ehrmann, M. Mitchell, P. Burgoyne, E. Simpson, and J. Dyson Identification of the Immunodominant HY H2-Dk Epitope and Evaluation of the Role of Direct and Indirect Antigen Presentation in HY Responses J. Immunol., December 1, 2005; 175(11): 7209 - 7217. [Abstract] [Full Text] [PDF]
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 P. R. Provost and Y. Tremblay Genes Involved in the Adrenal Pathway of Glucocorticoid Synthesis Are Transiently Expressed in the Developing Lung Endocrinology, May 1, 2005; 146(5): 2239 - 2245. [Abstract] [Full Text] [PDF]
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D. B. Miklos, H. T. Kim, K. H. Miller, L. Guo, E. Zorn, S. J. Lee, E. P. Hochberg, C. J. Wu, E. P. Alyea, C. Cutler, et al. Antibody responses to H-Y minor histocompatibility antigens correlate with chronic graft-versus-host disease and disease remission Blood, April 1, 2005; 105(7): 2973 - 2978. [Abstract] [Full Text] [PDF]
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 R. Ivanov, T. Aarts, S. Hol, A. Doornenbal, A. Hagenbeek, E. Petersen, and S. Ebeling Identification of a 40S Ribosomal Protein S4-Derived H-Y Epitope Able to Elicit a Lymphoblast-Specific Cytotoxic T Lymphocyte Response Clin. Cancer Res., March 1, 2005; 11(5): 1694 - 1703. [Abstract] [Full Text] [PDF]
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T. Higuchi, T. Maruyama, A. Jaramillo, and T. Mohanakumar Induction of Obliterative Airway Disease in Murine Tracheal Allografts by CD8+ CTLs Recognizing a Single Minor Histocompatibility Antigen J. Immunol., February 15, 2005; 174(4): 1871 - 1878. [Abstract] [Full Text] [PDF]
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H. Torikai, Y. Akatsuka, M. Miyazaki, E. H. Warren III, T. Oba, K. Tsujimura, K. Motoyoshi, Y. Morishima, Y. Kodera, K. Kuzushima, et al. A Novel HLA-A*3303-Restricted Minor Histocompatibility Antigen Encoded by an Unconventional Open Reading Frame of Human TMSB4Y Gene J. Immunol., December 1, 2004; 173(11): 7046 - 7054. [Abstract] [Full Text] [PDF]
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J.-G. Chai, E. James, H. Dewchand, E. Simpson, and D. Scott Transplantation tolerance induced by intranasal administration of HY peptides Blood, May 15, 2004; 103(10): 3951 - 3959. [Abstract] [Full Text] [PDF]
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 E. Zorn, D. B. Miklos, B. H. Floyd, A. Mattes-Ritz, L. Guo, R. J. Soiffer, J. H. Antin, and J. Ritz Minor Histocompatibility Antigen DBY Elicits a Coordinated B and T Cell Response after Allogeneic Stem Cell Transplantation J. Exp. Med., April 19, 2004; 199(8): 1133 - 1142. [Abstract] [Full Text] [PDF]
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D. B. Miklos, H. T. Kim, E. Zorn, E. P. Hochberg, L. Guo, A. Mattes-Ritz, S. Viatte, R. J. Soiffer, J. H. Antin, and J. Ritz Antibody response to DBY minor histocompatibility antigen is induced after allogeneic stem cell transplantation and in healthy female donors Blood, January 1, 2004; 103(1): 353 - 359. [Abstract] [Full Text] [PDF]
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N. Hirano, M. O. Butler, M. S. von Bergwelt-Baildon, B. Maecker, J. L. Schultze, K. C. O'Connor, P. H. Schur, S. Kojima, E. C. Guinan, and L. M. Nadler Autoantibodies frequently detected in patients with aplastic anemia Blood, December 15, 2003; 102(13): 4567 - 4575. [Abstract] [Full Text] [PDF]
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Y. Akatsuka, T. Nishida, E. Kondo, M. Miyazaki, H. Taji, H. Iida, K. Tsujimura, M. Yazaki, T. Naoe, Y. Morishima, et al. Identification of a Polymorphic Gene, BCL2A1, Encoding Two Novel Hematopoietic Lineage-specific Minor Histocompatibility Antigens J. Exp. Med., June 2, 2003; 197(11): 1489 - 1500. [Abstract] [Full Text] [PDF]
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M. Murata, E. H. Warren, and S. R. Riddell A Human Minor Histocompatibility Antigen Resulting from Differential Expression due to a Gene Deletion J. Exp. Med., May 19, 2003; 197(10): 1279 - 1289. [Abstract] [Full Text] [PDF]
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 W. A. E. Marijt, M. H. M. Heemskerk, F. M. Kloosterboer, E. Goulmy, M. G. D. Kester, M. A. W. G. van der Hoorn, S. A. P. van Luxemburg-Heys, M. Hoogeboom, T. Mutis, J. W. Drijfhout, et al. Hematopoiesis-restricted minor histocompatibility antigens HA-1- or HA-2-specific T cells can induce complete remissions of relapsed leukemia PNAS, March 4, 2003; 100(5): 2742 - 2747. [Abstract] [Full Text] [PDF]
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 A. J. Barrett, K. Rezvani, S. Solomon, A. M. Dickinson, X. N. Wang, G. Stark, H. Cullup, M. Jarvis, P. G. Middleton, and N. Chao New Developments in Allotransplant Immunology Hematology, January 1, 2003; 2003(1): 350 - 371. [Abstract] [Full Text] [PDF]
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 D. Contu, L. Morelli, P. Zavattari, R. Lampis, E. Angius, P. Frongia, D. Murru, M. Maioli, P. Francalacci, J. A. Todd, et al. Sex-Related Bias and Exclusion Mapping of the Nonrecombinant Portion of Chromosome Y in Human Type 1 Diabetes in the Isolated Founder Population of Sardinia Diabetes, December 1, 2002; 51(12): 3573 - 3576. [Abstract] [Full Text] [PDF]
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 E. James, D. Scott, J.-G. Chai, M. Millrain, P. Chandler, and E. Simpson HY peptides modulate transplantation responses to skin allografts Int. Immunol., November 1, 2002; 14(11): 1333 - 1342. [Abstract] [Full Text] [PDF]
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 X.-F. Yang, C. J. Wu, L. Chen, E. P. Alyea, C. Canning, P. Kantoff, R. J. Soiffer, G. Dranoff, and J. Ritz CML28 Is a Broadly Immunogenic Antigen, Which Is Overexpressed in Tumor Cells Cancer Res., October 1, 2002; 62(19): 5517 - 5522. [Abstract] [Full Text] [PDF]
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M. Guimond, A. Balassy, M. Barrette, S. Brochu, C. Perreault, and D. C. Roy P-glycoprotein targeting: a unique strategy to selectively eliminate immunoreactive T cells Blood, June 28, 2002; 100(2): 375 - 382. [Abstract] [Full Text] [PDF]
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M. H. J. Vogt, J. W. van den Muijsenberg, E. Goulmy, E. Spierings, P. Kluck, M. G. Kester, R. A. van Soest, J. W. Drijfhout, R. Willemze, and J. H. F. Falkenburg The DBY gene codes for an HLA-DQ5-restricted human male-specific minor histocompatibility antigen involved in graft-versus-host disease Blood, April 15, 2002; 99(8): 3027 - 3032. [Abstract] [Full Text] [PDF]
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V. T. Ho and R. J. Soiffer The history and future of T-cell depletion as graft-versus-host disease prophylaxis for allogeneic hematopoietic stem cell transplantation Blood, December 1, 2001; 98(12): 3192 - 3204. [Full Text] [PDF]
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 R. Nisini, G. Romagnoli, M. J. Gomez, R. La Valle, A. Torosantucci, S. Mariotti, R. Teloni, and A. Cassone Antigenic Properties and Processing Requirements of 65-Kilodalton Mannoprotein, a Major Antigen Target of Anti-Candida Human T-Cell Response, as Disclosed by Specific Human T-Cell Clones Infect. Immun., June 1, 2001; 69(6): 3728 - 3736. [Abstract] [Full Text] [PDF]
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R. F. Storb, R. Champlin, S. R. Riddell, M. Murata, S. Bryant, and E. H. Warren Non-Myeloablative Transplants for Malignant Disease Hematology, January 1, 2001; 2001(1): 375 - 391. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
3') used for generation of the
UTY minigenes are listed in Table
), or without HLA-B60 cDNA
(
). Forty-eight hours after transfection, HLA-B60 HY CTL was added.
Culture supernatants were harvested 1 day later and tested for TNF
production.
