| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
*
Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892; and
Cytel Corp., San Diego, CA 92121
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
We have recently shown that tumor Ags, TRP-1/gp75 and TRP-2, are recognized by the HLA-A31-restricted TIL586, which has been shown to result in tumor regression when infused along with IL-2 into the autologous patient with melanoma (15, 16). The gp75 epitope was unexpectedly found to be derived by translation of an alternative open reading frame of the gp75 gene, while the T cell epitope for TRP-2 was encoded by the normal open reading frame of the TRP-2 gene (20). Recognition of these antigenic peptides by TIL586 and its derived CTL clones was shown to be HLA-A31 restricted. Given the relatively low frequency of the HLA-A31 allele in major ethnic groups, broad utilization of these antigenic peptides in immunotherapy would be limited.
In the last few years, significant progress has been made toward understanding the rules governing peptide binding to MHC class I molecules, the so-called peptide binding motifs (21). Based on the structural similarities of a group of HLA alleles, peptide binding motifs, sequencing analysis of pools of naturally processed and endogenously bound peptides eluted from MHC class I molecules, and peptide binding assays, several supertypes were proposed: the HLA-A2-like, -A3-like, and -B7-like (22). The A3-like supertype includes the alleleic products of at least five of the most common HLA-A alleles: A3, A11, A31, A33, and A68.
In the present study we sought to expand the potential population coverage of TRP-1- and TRP-2-derived epitopes by testing the hypothesis that a particular peptide not only can bind to more than one type of HLA allele, but can still be recognized by CTL. As the first step, we demonstrate here that the antigenic peptides of TRP-1 and TRP-2 can bind to HLA-A3, -A11, -A31, -A33, and -A68. After screening 13 TILs (possibly restricted by HLA-A3, -A11 and -A33), one HLA-A33-restricted TIL1244 was identified that recognized the TRP197–205 in the context of both HLA-A31 and-A33, suggesting that a single TCR can recognize a tumor-specific self-peptide presented by two different HLA-A alleles. These findings indicate that the TRP-1 and TRP-2 antigenic peptides can be used for the development of peptide-based vaccines for the treatment of melanoma patients expressing not only HLA-A31, but also HLA-A33. Based on their good peptide binding affinities, it is also possible to use TRP197–205 or ORF3P to raise CTL from patients expressing HLA-A3, -A11, and -A68.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The following chemicals and reagents were purchased from the sources indicated: RPMI 1640 medium, AIM-V medium, Lipofectamine, and G418 from Life Technologies (Gaithersburg, MD); the eukaryotic expression vector pCR3 from Invitrogen (San Diego, CA); anti-HLA-A31 and anti-HLA-A33 mAbs from One Lambda (Canoga Park, CA); and anti-IgM Ab conjugated with FITC from Vector Laboratories, Inc. (Burlingame, CA).
T cell clones and lines
TIL586 and TIL1244 were isolated separately from tumor specimens of patients with metastatic melanoma and grown in medium containing IL-2 (6000 IU/ml; Chiron, Emeryville, CA) for 32 to 60 days as previously described (23). TIL586 and TIL1244 were predominantly CD8+ T cells. The T cell clones were generated by limiting dilution methods (at one cell per well) from the TIL1244 cell line using irradiated allogeneic PBL (1 x 103 cells/well) as feeder cells in RPMI 1640 medium containing 10% human AB serum and 500 IU of IL-2. After 12 days, T cell clones were expanded in AIM-V medium containing 6000 IU/ml IL-2. To obtain optimal expansion, we used the OKT3 expansion method described by S. Riddell (24). Briefly, on day 0, 5 x 104 to 5 x 105 T cells were cocultured with HLA-A33+ PBL (500:1, PBL:T cell ratio) and 1500EBV B cells (100:1, EBV:T cell ratio) in 25 ml of RPMI 1640 medium containing 11% human AB serum, 30 ng/ml OKT3 Ab, and antibiotics. On day 1, IL-2 was added to a final concentration of 180 IU/ml. On day 5, the cell culture was changed to fresh medium containing 11% human AB serum and 180 IU/ml of IL-2. The medium was then changed every 3 days. On days 12 to 14, T cells were harvested, counted, and cryopreserved.
Melanoma cell lines 397mel, 397mel/A31, 586mel, 624mel, and 624mel/A31; EBV-transformed B cell lines 586EBV and 1500EBV; and T2 cells were established in our laboratory and cultured in RPMI 1640 medium containing 10% FCS. 586EBV B cells are HLA-A31-positive and 1500EBV B cells are HLA-A33-positive cell lines. The COS-7 cell line was provided by Dr. W. Leonard (National Institutes of Health). The following EBV-transformed cell lines were used as sources of class I molecules: GM3107 (A*0301), BVR (A*1101), SPACH (A*3101), and LWAGS (A*3301). A CIR transfectant, characterized by Dr. Walter Stokus, was used as a source of A*6801. Cells were maintained in vitro by culture in RPMI 1640 medium supplemented with 2 mM L-glutamine and 10% heat-inactivated FCS. Cell cultures were also supplemented with 100 µg/ml of streptomycin (Irvine Scientific, Santa Ana, CA) and 100 U/ml of penicillin (Life Technologies, Grand Island, NY). Large quantities of cells were grown in spinner cultures.
GM-CSF secretion assay
DNA transfection and GM-CSF assays were performed as previously described (15). Briefly, 200 ng of DNA encoding Ags and 50 ng of HLA-A31 DNA were mixed with 2 µl of Lipofectamine in 100 µl of DMEM and incubated at room temperature for 15 to 45 min. The DNA/Lipofectamine mixture was then added to the COS-7 cells (5 x 104) and incubated overnight. The following day, cells were washed twice with DMEM medium. TIL586 was added at a concentration of 1 x 105 cells/well in AIM-V medium containing 120 IU/ml of IL-2. For T cell clones, only 1 to 2 x 104 cells/well were added. After an 18- to 24-h incubation, 100 µl of supernatant was collected, and GM-CSF was measured in a standard ELISA assay (R&D Systems, Minneapolis, MN). For peptide recognition, 586EBV or T2 cells were incubated with peptides at 37°C for 90 min and then washed three times with AIM-V medium containing 120 IU/ml of IL-2. T cells were added and incubated for an additional 18 to 24 h; 100 µl of supernatant was collected for the GM-CSF assay.
Cytotoxicity assays
The cytolytic assay was performed as previously described (7). Briefly, the target cells were labeled with chromium for 90 min. After washing three times, the cells were incubated with peptides at a concentration of 1 µg/ml for 90 min. The cells were washed again, counted, and then mixed with TIL1244, T cell clones, or CTL clone 4 at the indicated E:T cell ratio. Chromium release was measured after 4-h incubation. For titration of ORF3P and TRP197–205 peptides, 586EBV B cells and 1500EBV B cells were incubated with various concentrations of the purified peptide. The percentage of specific lysis was determined from the equation (A - B)/(C - B) x 100, where A is lysis of target cells by TIL1244 and T cell clones in the presence of a peptide, B is spontaneous release from EBV B cells in the presence of the same peptide but in the absence of effector cells, and C is the maximum chromium release.
The peptides were synthesized by a solid phase method using a peptide synthesizer (model AMS 422, Gilson Co., Inc., Worthington, OH). Some peptides were purified by HPLC and had >98% purity. The peptide mass of some peptides was confirmed by mass spectrometric analysis.
Affinity purification of HLA-A molecules
Cells were lysed at a concentration of 108 cells/ml in PBS containing 1% Nonidet P-40 and 1 mM PMSF. The lysates were cleared of debris and nuclei by centifugation at 10,000 x g for 20 min. MHC molecules were then purified by affinity chromatography as previously described (25). Columns of inactivated Sepharose CL4B and protein A-Sepharose were used as precolumns. Lysates were filtered through 0.8- and 0.4-µm filters and then depleted of HLA-B and HLA-C molecules by repeated passage over protein A-Sepharose beads conjugated with the anti-HLA(B,C) Ab B1.23.2 (26). Typically, two to four passages were required for effective depletion. Subsequently, the anti-HLA(A,B,C) Ab W6/32 (27) was used to capture HLA-A molecules.
Independently, both Ab columns were washed with 15 column volumes of 10
mM Tris in 1.0% Nonidet P-40, PBS, and 2 column volumes of PBS
containing 0.4% n-octylglucoside. Finally, the class I
molecules were eluted with 50 mM diethylamine in 0.15 M NaCl containing
0.4% n-octylglucoside, pH 11.5. A 1/25th volume of 2.0 M
Tris, pH 6.8, was added to the eluate to reduce the pH to 
8.0 and
then concentrated by centifugation in Centriprep 30 concentrators
(Amicon, Beverly, MA) at 2000 rpm. Protein purity, concentration, and
effectiveness of depletion steps were monitored by SDS-PAGE.
Class I peptide binding assays
Quantitative assays for the binding of three peptides to soluble
MHC class I molecules on the basis of the inhibition of binding of a
radiolabeled standard probe peptide to detergent solubilized MHC
molecules were conducted as previously described (28). Briefly,
purified human class I molecules (5–500 nM) were incubated with 1 to
10 nM 125I-radiolabeled probe peptide, iodinated by
the chloramine-T method (29), for 48 h at room temperature in the
presence of 1 µM human ß2m (Scripps Laboratories, San
Diego, CA) and a mixture of protease inhibitors. The final
concentrations of protease inhibitors were 1 mM PMSF, 1.3 nM 1.10
phenanthroline, 73 µM pepstatin A, 8 mM EDTA, and 200 µM
N
-p-tosyl-L-lysine
chloromethyl ketone (TLCK). Class I peptide complexes were separated
from free peptide by gel filtration on TSK200 columns, and the fraction
of bound peptide was calculated as previously described (25). In
preliminary experiments, the HLA class I prep was titrated in the
presence of fixed amounts of radiolabeled peptides to determine the
concentration of class I molecules necessary to bind 10 to 20% of the
total radioactivity. All subsequent inhibition and direct binding
assays were performed using these class I concentrations. In the
inhibition assays, peptide inhibitors were typically tested at
concentrations ranging from 120 µg/ml to 1.2 ng/ml. The data were
then plotted, and the dose yielding 50% inhibition was measured.
Peptides were tested in two to four completely independent experiments.
Since under these conditions concentration of label <
concentration of MHC and IC50 
concentration of MHC, the
measured IC50 values are reasonable approximations of the
true kilodalton values. The radiolabeled probe and standard control
peptides used are as follows. The A3CON1 peptide (sequence KVFPYALINK)
(30) was used as the radiolabeled probe for the A3, A11, A31, and
A*6801 assays. A T7Y analogue of HBVc141–151
(sequence STLPETYVVRR) was used as the radiolabeled probe for the
A*3301 assay.
The average IC50 values of A3CON1 for the A3, A11, A31, and A*6801 assays were 11, 6, 18, and 8 nM, respectively. The average IC50 of the HBVc141–151 peptide in the A*3301 assay was 29 nM.
RT-PCR analysis
RNA was extracted from T cells using the Trizol reagent according to manufacturer’s procedure (Life Technologies). T cells (1 x 107) of CTL clones 35 and 38 were used to isolate total RNA. RT-PCR was performed using the One-Step RT-PCR kit from Life Technologies and ßV subfamily-specific primer combined with a CßR primer from the constant region of a TCR as previously described (31). Two hundred nanograms of total RNA was used in a 50-µl RT-PCR reaction. RT-PCR was performed in one cycle of 94°C for 2 min and 50°C for 30 min, followed by 40 cycles of 94°C for 30 s for denature, 62°C for 20 s for annealing, and 72°C for 1 min for extension. The positive control used the constant region primers CßF and CßR. Negative controls used RNA or water instead of cDNA in the PCR reactions. RT-PCR products were resolved on a 1% agarose gel.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Based on the structural similarities and sequencing of pools of
naturally processed peptides bound on MHC class I molecules, it was
recently proposed that a set of HLA-A alleles, including HLA-A3, -A11,
-A31, -A33, and -A68, could be grouped into a superfamily or designated
the A3-like supertype (28). These HLA class I molecules bind to peptide
ligands with hydrophobic anchor residues at position 2 and positively
charged residues at the COOH-termini (21, 28) (Table I
). Two tumor Ag peptides, ORF3P, derived
from the gene product of alternative open reading frame of TRP-1, and
TRP197–205, derived from the TRP-2 normal coding
sequence, were recently identified. Both of these epitopes conformed to
the previously described canonical HLA-A31 binding motif (Table I). A
modified peptide of TRP197–205, TRP197–205K,
was also included in Table I, as this peptide was also recognized by
the HLA-A31-restricted CTL clone 4 when pulsed onto
HLA-A31+ EBV B cells. Since these tumor Ag peptides were
recognized by HLA-A31-restricted CTLs (16, 20), it was of interest to
test whether TRP-1 and TRP-2 could be recognized by CTLs in the context
of other HLA alleles, such as HLA-A3, -A11, -A33, and -A68. As the
first step, we performed peptide binding assays to determine whether
ORF3P, TRP197–205, and TRP197–205K could bind
to members of the A3-like family. As shown in Table II, these three peptides were indeed
capable of binding to A3, A11, A31, A33, and A68 molecules. The peptide
ORF3P bound to all five MHC class I members of the A3-like supertype
with relatively high binding affinity. The peptide
TRP197–205 exhibited a relatively high binding affinity to
HLA-A31, intermediate binding affinity to HLA-A33, and relatively low
binding affinity to HLA-A3, -A11, and -A68. Interestingly, the binding
affinity of TRP197–205 to HLA-A3 and A11 could be improved
significantly by the substitution of Arg with Lys at the C-terminus of
the peptide. As shown in Table II, TRP197–205K exhibits a
high binding affinity to both HLA-A3 and -A11, approximately 100-fold
higher than the parental peptide TRP197–205. Furthermore,
this substitution had little or no effect on the binding affinity of
the peptide to HLA-A31 and -A33, and only a marginally negative effect
on the binding affinity of the peptide to HLA-A68.
|
|
Although many tumor Ags have been identified and their T cell
epitopes have been determined (7), it has not been reported that the
same T cell epitope peptide can bind to different MHC alleles in the
superfamily and still be recognized by the corresponding CTL. To test
this possibility, we collected five TILs isolated from patients
expressing HLA-A3, six TILs from patients expressing HLA-A11, and two
TILs from patients expressing HLA-A33 and grew them in RPMI 1640 medium
with 10% human AB serum and 6000 IU/ml of IL-2. After 1 wk, these TILs
were tested for recognition of ORF3P and TRP197–205
pulsed on HLA-A3+, -A11+, and -A33+
EBV B cells, respectively. No T cell recognition was found from TILs
isolated from patients expressing either HLA-A3 or -A11. Failure to
identify HLA-A3- and HLA-A11-restricted TILs that recognize ORF3P or
TRP197–205 may be due to lack of true HLA-A3-
and HLA-A11-restricted CTL in the bulk TIL populations. However, one of
two HLA-A33-restricted TILs, TIL1244, was found to recognize the same
TRP197–205 peptide in the context of HLA-A33 (Table III). The phenotype of
HLA-A33+ 1500EBV was confirmed by FACS analysis. 1500EBV
stained positive with anti-HLA-A33 Ab, but negative with
anti-HLA-A31 Ab. By contrast, 586mel and 586EBV were positive for
HLA-A31 and negative for HLA-A33 (data not shown), suggesting that
HLA-A31 and -A33 molecules can be distinguished by mAbs.
|
Since TRP197–205 is a good HLA-A31 binder (Table II
), we next tested whether the HLA-A33-restricted TIL1244 was also
capable of recognizing the A31/peptide complex. To this end, ORF3P and
TRP197–205 were pulsed onto HLA-A31+ 586EBV,
HLA-A33+ 1500EBV, and T2 cells, respectively, and evaluated
for T cell recognition by measuring GM-CSF release. TIL1244 recognized
both HLA-A31+ and HLA-A33+ EBV B cells pulsed
with the TRP197–205 peptide, but not T2 cells
pulsed with the TRP197–205 peptide or HLA-A31+
or HLA-A33+ EBV B cells pulsed with the ORF3P peptide (Fig. 1). TIL1244 also recognized the modified
peptideTRP197–205K as effectively as the parental peptide.
In contrast, CTL clone 4, which recognized the TRP197–205
peptide pulsed onto HLA-A31+ EBV B cells, did not respond
to the TRP197–205 and TRP197–205K peptides
presented by HLA-A33+ EBV B cells (Fig. 1).
|
A). Interestingly,
TIL1244 also recognized COS-7 transfected with HLA-A31 plus TRP-2.
However, TIL1244 did not recognize COS-7 transfected with HLA-A31 plus
TRP-1 or with HLA-A31 alone (Fig. 2A). These results
demonstrated that TIL1244 were capable of recognizing the
TRP197–205 peptide presented by either HLA-A31 or HLA-A33
molecules.
|
B indicated that 586mel (A31+
TRP-2+), 397mel/A31 (A31+, TRP-2+),
and 624mel/A31 (A31+, TRP-2+) all stimulated
GM-CSF release from TIL1244, while 397mel (A31-,
TRP-2+) and 624mel (A31-, TRP-2+)
failed to stimulate cytokine release from TIL1244. In addition, 624mel
transfected with HLA-A33 was shown to confer recognition by TIL1244.
Taken together, these results indicated that TIL1244 is also capable of
recognizing tumor cells expressing TRP-2 and either HLA-A31 or
HLA-A33. T cell clones derived from TIL1244
Since TIL1244 is a bulk T cell line, recognition of
TRP197–205 by TIL1244 in the context of HLA-A31 and -A33
could be due to the coexistence of different subsets of T cell
populations: one recognizing TRP-2 in the context of HLA-A31 and the
other recognizing TRP-2 in the context of HLA-A33. To test this
possibility, T cell clones were generated by limiting dilution. Of 136
clones tested, 50 clones (36%) were found to recognize 586EBV B cells
and 1500EBV pulsed with TRP197–205, but not EBV B cells
alone. Four CTL clones were further expanded using the anti-OKT3
rapid expansion method and tested for their recognition of either
HLA-A31+ or HLA-A33+ tumor cells. All four CTL
clones recognized 586mel, 397mel/A31, 624mel/A31, and 624/A33 (data not
shown). The cytolytic activity of TIL1244 and its derived CTL clones
against different targets was also tested at different E:T ratios. As
shown in Figure 3, A and
B, TIL1244 efficiently lysed 586mel,
TRP197–205-pulsed 586EBV, and 1500EBV B cells, while no
lysis was observed in the case of 397mel or T2 cells pulsed with
TRP197–205. Similar results were obtained when the CTL
clones 35 and 38 were used as effector cells (Fig. 3, C and
D). The clonality of CTL clones 35 and 38 was
confirmed by RT-PCR analysis using TCR ßV subfamily-specific primers
(31). Only one DNA band of about 600 bp was detected from RT-PCR
products amplified with a ßV16-specific primer combined with a CßR
primer from the constant region of TCR, while no DNA band was observed
from the RT-PCR reactions using other ßV-specific primers combined
with the CßR primer (Fig. 4). These
results suggested that a single TCR receptor can recognize the same
peptide presented by either HLA-A31 or -A33 molecules.
|
|
From the experiments presented in Figure 3, it was evident that
lysis of 1500EBV pulsed with TRP197–205 peptide by
TIL1244 and CTL clones 35 and 38 was higher than that of 586EBV pulsed
with the same peptide. To further analyze the differential recognition
of target cells by both TIL1244 and its CTL clones, peptide titration
experiments were performed. As shown in Figure 5A, there was a significant
difference in the peptide concentrations required to obtain similar
cytokine release from the TIL1244 stimulated by different APCs. The T
cell response could be detected down to 1 ng/ml of
TRP197–205 and plateaued at 1 µg/ml in the case of
1500EBVB cells. However, at least a 10-fold higher peptide
concentration was required for detectable responses when the same
peptide was pulsed onto 586EBV B cells. The T cell response in this
case did not reach a plateau at a concentration of 1 µg/ml of the
peptide. Although the poor response of TIL1244 to the
TRP197–205 peptide when loaded on HLA-A31+
586EBV could result from the difference in peptide binding affinity to
HLA-A31 molecules on 586EBV vs that to HLA-A33 molecules on 1500EBV,
the peptide binding assay in Table II ruled out this possibility. In
contrast, the response of HLA-A31-restricted CTL clone 4 to the
TRP197–205 peptide reached a plateau at 1 ng/ml when
pulsed onto HLA-A31+ 586EBV B cells, and no recognition was
observed when the same peptide was pulsed onto HLA-A33+
1500EBV B cells (Fig. 5B). Furthermore, both 586EBV
and 1500EBV B cells expressed a comparable level of MHC class I
molecules (HLA-A31 or -A33) on the cell surface (data not shown). These
results indicated that the differential recognition of the same peptide
on either HLA-A31 or -A33 by TIL1244 might be due to the difference in
the TCR affinity of TIL1244 to A31/peptide compared with that to
A33/peptide. Consistent with this hypothesis were results
obtained in experiments in which various peptide concentrations were
evaluated (Fig. 6), strongly suggesting
that TCR affinity rather than MHC/peptide affinity is responsible for
the observed patterns of T cell recognition and target lysis.
|
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The binding affinity of TRP197–205 to HLA-A3 and -A11 can be improved by substituting Arg with Lys. It was reported that the improved binding affinity of the modified peptides by substitution of anchor residues with optimal or preferred residues enhanced the immunogenicity of peptides in vitro (32). However, the correlation between the peptide binding affinity and immunogenicity in vivo remains to be established. Recent reports suggest that peptide binding affinity for MHC class I molecules is a major factor, but not the sole factor, in determining immunogenicity (32, 33). Other factors, such as peptide liberation, TAP transport, and TCR repertoire for a particular peptide, also affect the immunogenicity of peptide in vivo (33). The existence of immunodominant peptides may affect the immunogenicity of the other peptides, such as a cryptic peptide (33, 34). For this reason, peptide-based vaccines may have advantages over other types of vaccines because one should be able to direct the immune response toward cryptic peptides in cases where an immunodominant epitope suppresses cryptic peptides to elicit an immune response (34).
Direct peptide binding assay has allowed the definition of several HLA supertypes initially suggested by HLA structural similarity and peptide binding motifs (22, 28). However, these studies did not demonstrate that these MHC/peptide complexes can be recognized by CTL. It was previously reported that a viral peptide of HBcAg from hepatitis B virus was capable of binding to HLA-A31 and -A68 and was recognized by their corresponding CTLs isolated from a single patient (35). To our knowledge, this is the first demonstration that a self cancer peptide, TRP197–205, not only binds to all members of the HLA-A3 superfamily, as discussed above, but is also recognized by the HLA-A31-restricted CTL clone 4 and the HLA-A33-restricted CTL derived from different patients. Furthermore, it was found that TIL1244 was capable of recognizing the TRP197–205 peptide presented by either HLA-A31 or -A33 molecules. These studies suggest that self Ags may be used to treat patients expressing HLA-A31 or HLA-A33 and perhaps other members of the HLA-A3-like supertype. Since the modified peptide TRP197–205K exhibits better binding to HLA-A3 and -A11, and still can be recognized by the HLA-A31-restricted CTL clone when pulsed onto A31+ EBV B cells as well as by TIL1244 when presented by both HLA-A31 and -A33 EBV B cells, this peptide may be a good candidate for a cancer vaccine for the treatment of patients expressing HLA-A3, -A11, -A31, and -A33. Generation of HLA-A3- or HLA-A11-restricted CTLs that are capable of recognizing these peptides derived from TRP-1 and TRP-2 is important for our understanding of T cell recognition of peptides presented by members of the A3 superfamily. These studies are currently under investigation.
Although the TRP197–205 peptide bound to HLA-A31 10 times
better than to HLA-A33 (Table II), recognition of the peptide/HLA-A31
complex by TIL1244 and its T cell clones was 10 times worse than that
of the peptide/HLA-A33 complex (Figs. 5 and 6), strongly suggesting
that the TCR receptor of TIL1244 or its T cell clones has a low avidity
for peptide/HLA-A31 complexes. Nonetheless, TIL1244 strongly recognized
both tumor cells expressing TRP-2 and HLA-A31 and tumor cells
expressing TRP-2 and HLA-A33 (Fig. 2B). This may be
due to high expression levels of TRP-2 in tumor cells (16), leading to
a high density of the TRP-2 peptide/HLA-A31 or HLA-A33 complexes on the
surface of tumor cells. T cell clones with differential TCR avidity for
antigenic peptide/MHC complexes have indeed recently been described
(36). Fully understanding the relationship between TCR avidity and the
number of MHC/peptides required for optimal TCR engagement will have
important implications for the treatment of patients with cancer or
autoimmune disease.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Rong-Fu Wang, Surgery Branch, National Cancer Institute, Building 10, Room 2B42, National Institutes of Health, Bethesda, MD 20892. E-mail address:
3 Abbreviations used in this paper: TRP, tyrosinase-related protein; TIL, tumor-infiltrating lymphocyte; ORF3P, open reading frame peptide-3; GM-CSF, granulocyte-macrophage colony-stimulating factor.
Received for publication May 21, 1997. Accepted for publication October 6, 1997.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
This article has been cited by other articles:
|
|
 |
|
  H. Liu, J. Fu, and M. Bouvier Allele- and Locus-Specific Recognition of Class I MHC Molecules by the Immunomodulatory E3-19K Protein from Adenovirus J. Immunol., April 1, 2007; 178(7): 4567 - 4575. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Leslie, D. A. Price, P. Mkhize, K. Bishop, A. Rathod, C. Day, H. Crawford, I. Honeyborne, T. E. Asher, G. Luzzi, et al. Differential Selection Pressure Exerted on HIV by CTL Targeting Identical Epitopes but Restricted by Distinct HLA Alleles from the Same HLA Supertype J. Immunol., October 1, 2006; 177(7): 4699 - 4708. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Lichterfeld, K. L. Williams, S. K. Mui, S. S. Shah, B. R. Mothe, A. Sette, A. Kim, M. N. Johnston, N. Burgett, N. Frahm, et al. T cell receptor cross-recognition of an HIV-1 CD8+ T cell epitope presented by closely related alleles from the HLA-A3 superfamily Int. Immunol., July 1, 2006; 18(7): 1179 - 1188. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Matsueda, H. Takedatsu, A. Yao, M. Tanaka, M. Noguchi, K. Itoh, and M. Harada Identification of Peptide Vaccine Candidates for Prostate Cancer Patients with HLA-A3 Supertype Alleles Clin. Cancer Res., October 1, 2005; 11(19): 6933 - 6943. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Paschen, M. Song, W. Osen, X. D. Nguyen, J. Mueller-Berghaus, D. Fink, N. Daniel, M. Donzeau, W. Nagel, H. Kropshofer, et al. Detection of Spontaneous CD4+ T-Cell Responses in Melanoma Patients against a Tyrosinase-Related Protein-2-Derived Epitope Identified in HLA-DRB1*0301 Transgenic Mice Clin. Cancer Res., July 15, 2005; 11(14): 5241 - 5247. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. A. Chianese-Bullock, J. Pressley, C. Garbee, S. Hibbitts, C. Murphy, G. Yamshchikov, G. R. Petroni, E. A. Bissonette, P. Y. Neese, W. W. Grosh, et al. MAGE-A1-, MAGE-A10-, and gp100-Derived Peptides Are Immunogenic When Combined with Granulocyte-Macrophage Colony-Stimulating Factor and Montanide ISA-51 Adjuvant and Administered as Part of a Multipeptide Vaccine for Melanoma J. Immunol., March 1, 2005; 174(5): 3080 - 3086. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. T. Loffredo, J. Sidney, C. Wojewoda, E. Dodds, M. R. Reynolds, G. Napoe, B. R. Mothe, D. H. O'Connor, N. A. Wilson, D. I. Watkins, et al. Identification of Seventeen New Simian Immunodeficiency Virus-Derived CD8+ T Cell Epitopes Restricted by the High Frequency Molecule, Mamu-A*02, and Potential Escape from CTL Recognition J. Immunol., October 15, 2004; 173(8): 5064 - 5076. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Azuma, T. Sasada, H. Takedatsu, H. Shomura, M. Koga, Y. Maeda, A. Yao, T. Hirai, A. Takabayashi, S. Shichijo, et al. Ran, a Small GTPase Gene, Encodes Cytotoxic T Lymphocyte (CTL) Epitopes Capable of Inducing HLA-A33-restricted and Tumor-Reactive CTLs in Cancer Patients Clin. Cancer Res., October 1, 2004; 10(19): 6695 - 6702. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Sasada, H. Takedatsu, K. Azuma, M. Koga, Y. Maeda, S. Shichijo, H. Shoumura, T. Hirai, A. Takabayashi, and K. Itoh Immediate Early Response Gene X-1, a Stress-Inducible Antiapoptotic Gene, Encodes Cytotoxic T-Lymphocyte (CTL) Epitopes Capable of Inducing Human Leukocyte Antigen-A33-Restricted and Tumor-Reactive CTLs in Gastric Cancer Patients Cancer Res., April 15, 2004; 64(8): 2882 - 2888. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Takedatsu, S. Shichijo, K. Katagiri, H. Sawamizu, M. Sata, and K. Itoh Identification of Peptide Vaccine Candidates Sharing among HLA-A3+, -A11+, -A31+, and -A33+ Cancer Patients Clin. Cancer Res., February 1, 2004; 10(3): 1112 - 1120. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Sidney, S. Southwood, V. Pasquetto, and A. Sette Simultaneous Prediction of Binding Capacity for Multiple Molecules of the HLA B44 Supertype J. Immunol., December 1, 2003; 171(11): 5964 - 5974. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. R. Burrows, R. A. Elkington, J. J. Miles, K. J. Green, S. Walker, S. M. Haryana, D. J. Moss, H. Dunckley, J. M. Burrows, and R. Khanna Promiscuous CTL Recognition of Viral Epitopes on Multiple Human Leukocyte Antigens: Biological Validation of the Proposed HLA A24 Supertype J. Immunol., August 1, 2003; 171(3): 1407 - 1412. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Negroiu, R. A. Dwek, and S. M. Petrescu The Inhibition of Early N-Glycan Processing Targets TRP-2 to Degradation in B16 Melanoma Cells J. Biol. Chem., July 11, 2003; 278(29): 27035 - 27042. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Guan, I. A. Doytchinova, and D. R. Flower HLA-A3 supermotif defined by quantitative structure-activity relationship analysis Protein Eng. Des. Sel., January 1, 2003; 16(1): 11 - 18. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Ueno, H. Tomiyama, and M. Takiguchi Single T Cell Receptor-Mediated Recognition of an Identical HIV-Derived Peptide Presented by Multiple HLA Class I Molecules J. Immunol., November 1, 2002; 169(9): 4961 - 4969. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Huarte, P. Sarobe, J. Lu, N. Casares, J. J. Lasarte, J. Dotor, M. Ruiz, J. Prieto, E. Celis, and F. Borras-Cuesta Enhancing Immunogenicity of a CTL Epitope from Carcinoembryonic Antigen by Selective Amino Acid Replacements Clin. Cancer Res., July 1, 2002; 8(7): 2336 - 2344. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. T. Khong and S. A. Rosenberg Pre-Existing Immunity to Tyrosinase-Related Protein (TRP)-2, a New TRP-2 Isoform, and the NY-ESO-1 Melanoma Antigen in a Patient with a Dramatic Response to Immunotherapy J. Immunol., January 15, 2002; 168(2): 951 - 956. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
