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The Journal of Immunology, 2000, 164: 4130-4134.
Copyright © 2000 by The American Association of Immunologists

A Human CTL Recognizes a Caspase-8-Derived Peptide on Autologous HLA-B*3503 Molecules and Two Unrelated Peptides on Allogeneic HLA-B*3501 Molecules1

Susanna Mandruzzato2, Vincent Stroobant, Nathalie Demotte and Pierre van der Bruggen4

Ludwig Institute for Cancer Research, Brussels Branch, and Unité de Génétique Cellulaire, Université Catholique de Louvain, Brussels, Belgium


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results and Discussion
 References
 
A CTL clone that recognizes autologous tumor cells was previously isolated from the blood of a head-and-neck cancer patient. The Ag was identified as peptide FPSDSWCYF presented by autologous HLA-B*3503 molecules. This peptide was encoded by a mutated CASP-8 gene, which is implicated in the triggering of apoptosis. Here, we show that this CTL clone, which expresses a single TCR, also recognizes two unrelated peptides on allogeneic HLA-B*3501 molecules. One peptide, HIPDVITY, is encoded by squalene synthase, and the other one, QFADVIVLF, is encoded by 2-hydroxyphytanoyl-CoA lyase. Both genes are expressed ubiquitously. These antigenic peptides are processed and presented by HLA-B*3501 cells. The two HLA-B35 alleles are closely related. Our results might reinforce the notion that the recognition of allogeneic HLA molecules depends on the presence in their groove of a limited number of peptides processed from ubiquitous proteins.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results and Discussion
 References
 
The interaction between the TCR and its ligand involves a molecule of the MHC and its associated peptide (1). While previous models supported the notion that T cells are specific for a unique ligand, many more recent studies, using analogues of immunogenic peptides, have demonstrated flexibility in TCR recognition (1). This notion of flexibility has also been extended to the phenomenon of alloreactivity: it is generally agreed that alloreactivity is the result of cross-reactive recognition by self-MHC-restricted T cells rather than a property of a particular population of T cells (2). However, the assertion that a peptide has a role in allorecognition remains controversial (3).

Very few data support this notion. In particular, only two examples of CTL clones have been reported that recognize both a peptide presented by a self-MHC class I molecule and other peptides in association with an allo-MHC. One is the murine CTL clone 2C, which was isolated from a H-2b BALB/b mouse injected with cells from a H-2d DBA/2 mouse (4). This CTL recognizes a peptide on allo-H-2 Ld molecules and another peptide on allo-H-2 Kbm3 molecules (Table IGo). By screening a random peptide library with clone 2C in the context of the self-Kb molecules, a third peptide, SIYRYYGL, was identified (5). It is not known whether this third peptide is a naturally processed epitope.


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  		Table I. Defined peptide-MHC ligands for three crossreactive CTL clones

 
The second example is a human CTL clone directed against an epitope from the EBV, which is presented by autologous HLA-B8 (6). This clone was also stimulated by HLA-B35 cells. Burrows and colleagues tested analogues of the viral peptides to identify the amino acid changes tolerated by the CTL (7). This information, in combination with a peptide binding motif for HLA-B35, was used to search a human protein sequence database. This revealed two distinct peptides that were recognized by the CTL when presented by allo-HLA-B35 molecules (Table IGo). Two of the peptide sequences are derived from ubiquitously expressed proteins. However, there is no information concerning the presence of the processed epitope on HLA-B35 target cells.

In a previous study, we described a CTL clone that lysed autologous cells of a squamous carcinoma of the oral cavity (8). The Ag was identified as peptide FPSDSWCYF, which was presented by autologous HLA-B*3503 molecules. This peptide was encoded by a mutated CASP-8 gene, which is implicated in the triggering of apoptosis. Here, we describe the identification of two unrelated peptides, which are processed from ubiquitous proteins and are recognized on allogeneic HLA-B*3501 molecules by this CTL clone.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results and Discussion
 References
 
Cell lines

Culture media were supplemented with 10% FCS and with L-arginine (116 mg/ml), L-asparagine (36 mg/ml), L-glutamine (216 mg/ml), streptomycin (0.1 mg/ml) and penicillin (200 U/ml). All the cultures were maintained at 37°C and in 8% CO2. Squamous cell carcinoma cell line BB49-SCCHN was adapted to culture from a tumor mass resected from the floor of the oral cavity of patient BB49, a 70-year old Caucasian woman in stage IV (8). BB49-SCCHN was grown in Iscove’s medium (Life Technologies, Gaithersburg, MD) supplemented with ACL-4 (9). COS-7 cells were grown in DMEM (Life Technologies). Cell lines BB49-EBV, LG-2-EBV, and the .221 cell lines transfected with HLA-B*3501 or HLA-B*3503 were grown in Iscove’s medium (10, 11). CTL clone 121 was maintained by weekly restimulation of 5 x 105 CTL with 105 irradiated BB49-SCCHN.B7 cells (BB49-SCCHN transfected with B7), pretreated for 72 h with 100 U/ml of IFN-{gamma}, in 2 ml of Iscove’s medium containing 10% human serum, 50 U/ml of IL-2, 0.8 x 106 irradiated LG-2-EBV cells, and 1.5 x 106 allogeneic irradiated PBMC.

TNF assay

A total of 1500 CTL were added in flat-bottom microwells containing 104 target cells in a total volume of 100 µl of Iscove’s medium containing 10% human serum and 25 U/ml of IL-2. After 4 or 24 h, the supernatant was collected, and its TNF content was determined by testing its cytotoxic effect on cells of WEHI-164 clone 13 (12) in a MTT colorimetric assay (13, 14).

Production of progressive deletions in cDNA 668

To generate progressive deletions from the 3' end of cDNA 668, and thereby obtain a large number of truncated cDNA clones, we used the Erase-a-base System (Promega, Madison, WI) as described (15).

Antigenic peptides and CTL assay

Peptides were synthesized on solid phase using F-moc for transient NH2-terminal protection as described (16) and were characterized using mass spectrometry. All peptides were >80% pure, as indicated by analytical HPLC. Lyophilized peptides were dissolved at 20 mg/ml in DMSO, diluted at 2 mg/ml in 10 mM acetic acid, and stored at -20°C. Peptides were tested in chromium release assays where 1,000 51Cr-labeled target cells were incubated for 30 min at room temperature in 96-well microplates with various concentrations of peptide before adding an equal volume containing 10,000 CTL 121. The indicated concentrations of peptide represent the final concentrations during the incubation of the target cells with the CTL. The assay was terminated after 4 h of incubation at 37°C.

RNA extraction, cDNA synthesis, and PCR amplification of TCRV{alpha} and TCRVß

Total RNA was prepared from CTL clone 121 using TRIZOL (Life Technologies), and first strand cDNA was synthetized with oligo(dT) and reverse transcriptase (Life Technologies). PCR was conducted by amplification with primers complementary to TCR V and C region sequences (17) in a 25-µl reaction mixture containing 2.5 µl of cDNA, all four dNTPs (each at 0.2 mM), 0.8 µM of each primer, and 0.5 U of Dynazyme (Finnzymes Oy, Finland) on a DNA thermal cycler (Biometra, Thermotrioblock, Westburg, The Netherlands). Amplification was performed for 25 cycles to amplify TCR{alpha} and 28 cycles to amplify TCRß, each consisting of 1 min at 94°C, 1 min at 60°C, and 1 min at 72°C. The PCR product of TCRß was cloned into the pTZ18R vector and sequenced by the dideoxy-chain termination method (ThermosequenaseTM cycle sequencing kit; Amersham, Little Chalfont, U.K.). The PCR products of TCR{alpha} were directly sequenced by the dideoxy-chain termination method (ThermosequenaseTM terminator cycle sequencing kit; Amersham). Nucleotide sequences were compared with available published TCR sequences (18, 19, 20, 21). The TCR nomenclature proposed by the International Union of Immunological Societies was adopted (22).

Peptide binding assay

We designed a peptide binding assay similar to the assay on intact human B cells described by van der Burg and colleagues (23). Peptides were stripped by mild acid treatment (pH 2.9), after which cells were incubated overnight at 4°C with a fluorescein (FL)4-labeled reference peptide at a fixed concentration (250 nM), together with decreasing concentrations (50 µM to 0.18 µM) of the three peptides of interest. After washing, the effectiveness by which these peptides competed for binding to HLA molecules was assayed by measuring the amount of HLA-bound FL-labeled peptide with FACscan analysis. The reference peptide was LPSC(FL)ADVEF, a Cys-derivative of the tyrosinase antigenic peptide presented by HLA-B35, LPSSADVEF (24).


   Results and Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
References
 
By stimulating blood cells of patient BB49 with irradiated autologous tumor cells, we isolated CD8+ CTL clone 121. To identify the Ag recognized by this CTL, a cDNA library was prepared with RNA from the autologous tumor cells. DNA was extracted from pools of 100 recombinant clones and transfected into COS-7 cells together with a cDNA coding for allogeneic HLA-B*3501 molecules. Patient BB49 was typed HLA-B*3503 and not B*3501, but for convenience we used an HLA-B*3501 cDNA, which was already cloned in an appropriate expression vector. The only difference between allo-B*3501 and self-B*3503 molecules is a S to F change at position 116 in the F pocket of the HLA molecule, which is the pocket that interacts with the C terminus of the peptide (25, 26). The transfected cells were screened for the expression of the Ag by adding CTL 121 and then measuring TNF production after 1 day of coculture. Of the 800 cDNA pools that were tested, four proved positive. Bacteria from the four positive pools were subcloned, and in each of them we obtained several clones that transferred the expression of the Ag in cells expressing allo-B*3501 molecules. Results obtained with a representative clone from each pool are shown in Fig. 1Go. When these four positive cDNA clones were transfected into COS-7 cells together with a cDNA coding for self-B*3503 molecules, only cDNA 668 was able to transfer the expression of the Ag (Fig. 1Go). The coding sequence of cDNA clone 668 was found to be identical, with the exception of a mutation, to that of CASP-8 coding for caspase-8, which is implicated in the triggering of apoptosis (8). The antigenic peptide is FPSDSWCYF, which is not encoded by the wild-type CASP-8. It produced half-maximal lysis of autologous EBV-B target cells at ~4 nM and on .221 cells expressing self-B*3503 molecules at ~1 nM (Fig. 2Go).



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  		FIGURE 1. Stimulation of CTL 121 by COS-7 cells transfected with cDNA clones isolated from a library together with sequences coding for allo-HLA-B*3501 or self-HLA-B*3503 molecules. A cDNA library was constructed with RNA extracted from tumor cells of patient BB49. A total of 20,000 COS-7 cells were cotransfected with an HLA cDNA and each of the four cDNA clones isolated from this library. One day after transfection, 1500 CTL 121, isolated from patient BB49, were added into microwells containing the transfected cells. Nonrelevant cDNA clone F10 was used as a negative control for stimulation. Autologous BB49 tumor cells were used as positive control for stimulation. TNF production was estimated after overnight coculture by testing the toxicity of the supernatants for the TNF-sensitive WEHI-164 clone 13 cells.

 


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  		FIGURE 2. Titration analysis of peptides recognized by CTL 121. Target cells were the autologous EBV-transformed B cell line (BB49-EBV) and .221 cells transfected with a cDNA coding for allo-HLA-B*3501 or self HLA-B*3503 molecules. A total of 1,000 chromium-labeled target cells were incubated at room temperature for 30 min with the indicated peptides at various concentrations before adding an equal volume containing 10,000 CTL 121. The indicated concentrations of peptide represent the final concentrations during the incubation of the target cells with the CTL. Chromium release was measured after 4 h. 2-HPCL, 2-hydroxyphytanoyl-CoA lyase.

 
The other cDNAs that code for an antigenic peptide presented to CTL 121 by allo-B*3501 molecules were also analyzed. cDNA 261 corresponds to squalene synthase (accession no. X69141), which is expressed in normal tissues and implicated in the synthesis of sterols (27). To identify the region of squalene synthase that codes for the peptide, cDNA 261 was digested with exonuclease III to generate progressive deletions from the 3' end of the coding sequence. To test their ability to code for the Ag, the truncated cDNA clones were transfected into COS-7 together with an allo-B*3501 cDNA, and the transfected cells were tested for their ability to stimulate the CTL. A short region, where the C terminus of the antigenic peptide had to be encoded, was delineated by comparing the sequence of the longest negative and the shortest positive cDNA. Synthetic peptides corresponding to the sequence of this region were synthesized and tested for recognition on .221 cells expressing allo-B*3501 molecules. The shortest antigenic peptide was an octamer, HIPDVITY (amino acids 269–276). Half-maximal lysis was obtained at a peptide concentration of ~10 nM (Fig. 2Go).

cDNAs 167 and 576 correspond to 2-hydroxyphytanoyl-CoA lyase (accession no. HSA131753), which is also expressed in normal tissues and catalyzes the carbon-carbon bond cleavage during oxidation of some fatty acids (28). The peptide recognized by CTL 121 was identified with the same strategy as described for squalene synthase. The shortest antigenic peptide was a nonamer, QFADVIVLF (amino acids 177–185). Half-maximal lysis of .221 cells expressing allo-B*3501 molecules was obtained at a peptide concentration of ~1 nM (Fig. 2Go).

Each of the three peptides were titrated for recognition by CTL 121 on autologous EBV-B cells, .221 cells expressing allo-B*3501 molecules, or .221 cells expressing self-B*3503 molecules (Fig. 2Go). On the autologous EBV-B cell line, only the peptide derived from mutated CASP-8 resulted in recognition. Surprisingly, the squalene synthase peptide, HIPDVITY, was also recognized by the CTL on self-B*3503 molecules, whereas COS-7 cells transfected with the squalene synthase cDNA and a HLA-B*3503 cDNA were not (Fig. 1Go). But the amount of peptide HIPDVITY needed to induce half-maximal lysis was ~10 nM, whereas only ~1 nM of the CASP-8-derived peptide was necessary. On .221 cells expressing allo-B*3501 molecules, the three peptides were recognized, as expected, with half-maximal lysis obtained at a peptide concentration of <10 nM.

To determine whether the affinity of each of the three peptides for the two HLA-B35 subtypes influenced the recognition by the CTL, we designed a peptide binding assay similar to the assay on intact human B cells described by van der Burg and colleagues (23). Briefly, the natural peptides were stripped from the HLA class I molecules by mild acid treatment, after which the .221.B*3501 or .221.B*3503 cells were incubated overnight at 4°C with a FL-labeled reference peptide at a fixed concentration, together with decreasing concentrations of the three peptides of interest. After washing, the effectiveness by which these peptides competed for binding to HLA molecules was assayed by measuring the amount of HLA-bound FL-labeled peptide with FACscan analysis. The concentrations needed to inhibit the binding of the FL-labeled peptide to 50% (IC 50) are indicated in Table IIGo. Peptide HIPDVITY did not bind to self-B*3503 molecules, thus revealing that it was not recognized by CTL clone 121. For the two other peptides, the amount needed to induce lysis was correlated to their affinity to self-B*3503. Concerning allo-B*3501 molecules, the three peptides can be considered to have a high affinity; the mutated peptide was the best competitor. Their recognition was not strictly correlated with their affinity.


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  		Table II. Binding capacity of the three peptides on each HLA-B35 subtype

 
The two peptides recognized on allo-B*3501 molecules only partially match the HLA-B35 binding motif, i.e., P in position 2 and Y, F, M, L, or I at the C terminus (29). They share amino acids DVI in the center of the peptide (Table IGo). Interestingly, only a D at position 4 is shared by the three peptides recognized by CTL 121. This amino acid seems to be essential for recognition, because replacing in each of the three peptides the D at position 4 by an A prevents recognition by the CTL (data not shown).

We verified that CTL 121 expresses only one TCR. RNA was prepared from CTL 121 and RT-PCR amplification was conducted with primers complementary to the V and C regions of the TCR. Sequencing revealed that CTL 121 expresses Vß14 and two V{alpha} transcripts, V{alpha}27 and V{alpha}9 (Fig. 3Go). The V{alpha}9 transcript is not functional because the sequences of the J and C regions are not in frame. Thus, CTL 121 expresses only one TCR. Taking into account the conserved conformation of the CDR1{alpha} loops that have been studied so far by crystallography (30), the CDR1{alpha} amino acid sequence of the TCR of CTL 121 was compared with the sequence of TCR A6 interacting with a tax peptide presented by HLA-A2. The crystal structure of this last TCR had revealed that it was the Q at position 30 of the CDR1{alpha} that interacts with a neutral G at position 4 of the peptide (31). Interestingly, in the TCR of CTL 121, there is a positively charged R at position 30, which may interact with the negatively charged D, which seemed to be essential for recognition and was present at position 4 of each of the three peptides recognized by CTL 121.



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  		FIGURE 3. Nucleotide and amino acid sequences of the TCR of CTL clone 121. Only the last three residues of the V region are shown, followed by the CDR3 region, the J region, and by the first three residues of the C region. TCRAJ segments were assigned according to Koop et al. (20 ), and TCRBD, BJ, and BC elements according to Toyonaga et al. (21 ). The TCRAV and TCRBV CDR3 length are defined according to Moss and Bell (36 ). TCR joining segment residues contributing to CDR3 are underlined.

 
The evidence that the peptides are processed was first obtained by transfection into COS-7 cells. We also tested two tumor cell lines obtained from B*3501 patients for their ability to stimulate CTL 121, because a unusually high expression can be achieved in transiently transfected COS-7 cells. The tumor cell lines scored positive, but the production of TNF by the CTL was not very high (Fig. 4Go). A bladder carcinoma line, transiently transfected with an allo-B*3501 cDNA, also stimulated the CTL to produce TNF (Fig. 4Go). Most probably, the number of peptide-HLA complexes was too low to result in maximal stimulation. This is in accordance with the 10–15% lysis of the .221 cells transfected with allo-B*3501 that was observed in absence of synthetic peptide (Fig. 2Go).



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  		FIGURE 4. Stimulation of CTL 121 by HLA-B35 tumor cell lines. The tumor cell lines were renal cell carcinoma lines LE9211 and LB1047, HLA-B35 negative bladder carcinoma line LB1831, autologous head-and-neck squamous carcinoma line BB49, and melanoma line LG2. Bladder carcinoma line LB831 was transiently transfected with an HLA-B*3501 cDNA, obtained by RT-PCR from the RNA of LB1047 and cloned into expression vector pcDNAI/Amp. A total of 1,500 CTL were added into microwells containing 10,000 tumor cell lines. The production of TNF was measured after 24 h of coculture by testing the toxicity of the supernatants for the TNF-sensitive WEHI-164.13 cells.

 
Whether or not the recognition of allogeneic HLA molecules by CTL is dependent on the presence of a peptide remains a subject of debate (3, 4, 32, 33). Here we show that CTL 121 clearly required the presence of a peptide for recognition and, moreover, that only a limited number of peptides can activate the TCR of clone 121. In fact, we have screened 80,000 cDNA with CTL 121 and only two different cDNAs were found to encode a peptide presented to CTL 121 on allo-B*3501 molecules. We are fully aware that the two HLA alleles are closely related and that one should be circumspect about the value of a generalized conclusion. However, it has already been reported that a single amino acid mismatch between two HLA class I molecules can induce a T cell response (34). Taken together, our results might reinforce the notion that peptides recognized on allogeneic MHC molecules are processed from ubiquitous proteins (32, 35).


   Acknowledgments
 
We gratefully acknowledge the excellent assistance of C. Wildmann. We are grateful to Dr. Soo Young Yang (Sloan-Kettering, New York, NY) for her assistance in HLA typing of patient BB49 and for providing cell lines. Many thanks to Dr. K. J. Smith and Prof. D. C. Wiley (Harvard University, Cambridge, MA) for helpful discussions and suggestions. We thank our colleagues Drs. A. Van Pel, P. Coulie, and B. Van den Eynde, for critical reading of the manuscript. We also thank M. Gandolfi and S. Mapp for help in the manuscript preparation.


   Footnotes
 
1 S.M. was supported by a postdoctoral fellowship from the "Training and Mobility of Researchers" program of the European Commission and by the International Center for Genetic Engineering and Biotechnology, Trieste, Italy. Back

2 Current address: Department of Oncology and Surgical Sciences, Oncology Section, via Gattamelata 64, 35128 Padova, Italy. Back

3 Address correspondence and reprint requests to Dr. P. van der Bruggen, Ludwig Institute for Cancer Research, Avenue Hippocrate 74, Université Catholique de Louvain 74.59, B-1200 Brussels, Belgium. Back

4 Abbreviation used in this paper: FL, fluorescein. Back

Received for publication July 1, 1999. Accepted for publication February 1, 2000.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 References
 

  1. Mason, D.. 1998. A very high level of crossreactivity is an essential feature of the T-cell receptor. Immunol. Today 19:395.[Medline]
  2. Sherman, L. A., S. Chattopadhyay. 1993. The molecular basis of allorecognition. Annu. Rev. Immunol. 11:385.[Medline]
  3. Smith, P. A., A. Brunmark, M. R. Jackson, T. A. Potter. 1997. Peptide-independent recognition by alloreactive cytotoxic T lymphocytes (CTL). J. Exp. Med. 185:1023.[Abstract/Free Full Text]
  4. Udaka, K., T. J. Tsomides, H. N. Eisen. 1992. A naturally occurring peptide recognized by alloreactive CD8+ cytotoxic T lymphocytes in association with a class I MHC protein. Cell 69:989.[Medline]
  5. Udaka, K., K. H. Wiesmüller, S. Kienle, G. Jung, P. Walden. 1996. Self-MHC-restricted peptides recognized by an alloreactive T lymphocyte clone. J. Immunol. 157:670.[Abstract]
  6. Burrows, S. R., S. J. Rodda, A. Suhrbier, H. M. Geysen, D. J. Moss. 1992. The specificity of recognition of a cytotoxic T lymphocyte epitope. Eur. J. Immunol. 22:191.[Medline]
  7. Burrows, S. R., S. L. Silins, R. Khanna, J. M. Burrows, M. Rischmueller, J. McCluskey, D. J. Moss. 1997. Cross-reactive memory T cells for Epstein-Barr virus augment the alloresponse to common human leukocyte antigens: degenerate recognition of major histocompatibility complex-bound peptide by T cells and its role in alloreactivity. Eur. J. Immunol. 27:1726.[Medline]
  8. Mandruzzato, S., F. Brasseur, G. Andry, T. Boon, P. van der Bruggen. 1997. A CASP-8 mutation recognized by cytolytic T lymphocytes on a human head and neck carcinoma. J. Exp. Med. 186:785.[Abstract/Free Full Text]
  9. Gazdar, A. F., H. K. Oie. 1986. Re: Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serum-free defined medium. Cancer Res. 46:6011.[Free Full Text]
  10. Shimizu, Y., R. DeMars. 1989. Production of human cells expressing individual transferred HLA-A,-B,-C genes using an HLA-A,-B,-C null human cell line. J. Immunol. 142:3320.[Abstract]
  11. Ragupathi, G., N. Cereb, S. Y. Yang. 1995. The relative distribution of B35 alleles and their IEF isotypes in a HLA-B35-positive population. Tissue Antigens 46:24.[Medline]
  12. Espevik, T., J. Nissen-Meyer. 1986. A highly sensitive cell line, WEHI 164 clone 13, for measuring cytotoxic factor/tumor necrosis factor from human monocytes. J. Immunol. Methods 95:99.[Medline]
  13. Hansen, M. B., S. E. Nielsen, K. Berg. 1989. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J. Immunol. Methods 119:203.[Medline]
  14. Traversari, C., P. van der Bruggen, B. Van den Eynde, P. Hainaut, C. Lemoine, N. Ohta, L. Old, T. Boon. 1992. Transfection and expression of a gene coding for a human melanoma antigen recognized by autologous cytolytic T lymphocytes. Immunogenetics 35:145.[Medline]
  15. Boël, P., C. Wildmann, M.-L. Sensi, R. Brasseur, J.-C. Renauld, P. Coulie, T. Boon, P. van der Bruggen. 1995. BAGE, a new gene encoding an antigen recognized on human melanomas by cytolytic T lymphocytes. Immunity 2:167.[Medline]
  16. Atherton, E., C. J. Logan, R. C. Sheppard. 1981. Peptide synthesis. II. Procedures for solid phase synthesis using N{alpha}-fluorenylmethysoxycarbamylamino-acid on polymide supports: synthesis of substance P and of acyl carrier protein 65–74 decapeptide. J. Chem. Soc. Lond. Perkin Trans. 1:538.
  17. Genevée, C., A. Diu, J. Nierat, A. Caignard, P. Y. Dietrich, L. Ferradini, S. Roman-Roman, F. Triebel, T. Hercend. 1992. An experimentally validated panel of subfamily-specific oligonucleotide primers (V{alpha}1-w29/Vß1-w24) for the study of human T cell receptor variable V gene segment usage by polymerase chain reaction. Eur. J. Immunol. 22:1261.[Medline]
  18. Yoshikai, Y., S. P. Clark, S. Taylor, U. Sohn, B. I. Wilson, M. D. Minden, T. W. Mak. 1985. Organization and sequences of the variable, joining and constant region genes of the human T-cell receptor {alpha}-chain. Nature 316:837.[Medline]
  19. Arden, B., S. P. Clark, D. Kabelitz, T. W. Mak. 1995. Human T-cell receptor variable gene segment families. Immunogenetics 42:455.[Medline]
  20. Koop, B. F., L. Rowen, K. Wang, C. L. Kuo, D. Seto, J. A. Lenstra, S. Howard, W. Shan, P. Deshpande, L. Hood. 1994. The human T-cell receptor TCRAC/TCRDC (C{alpha}/C{delta}) region: organization, sequence, and evolution of 97.6 kb of DNA. Genomics 19:478.[Medline]
  21. Toyonaga, B., Y. Yoshikai, V. Vadasz, B. Chin, T. W. Mak. 1985. Organization and sequences of the diversity, joining, and constant region genes of the human T-cell receptor ß chain. Proc. Natl. Acad. Sci. USA 82:8624.[Abstract/Free Full Text]
  22. Williams, A. F., J. L. Strominger, J. Bell, T. W. Mak, J. Kappler, P. Marrack, B. Arden, M. P. Lefranc, L. Hood, S. Tonegawa, M. Davis. 1993. Nomenclature for T-cell receptor (TCR) gene segments of the immune system. Bull. W. H. O. 71:113.[Medline]
  23. van der Burg, S. H., E. Ras, J. W. Drijfhout, W. E. Benckhuijsen, A. J. Bremers, C. J. Melief, W. M. Kast. 1995. An HLA class I peptide-binding assay based on competition for binding to class I molecules on intact human B cells. Identification of conserved HIV-1 polymerase peptides binding to HLA-A*0301. Hum. Immunol. 44:189.[Medline]
  24. Morel, S., A. Ooms, A. Van Pel, T. Wolfel, V. Brichard, P. van der Bruggen, B. J. Van den Eynde, G. Degiovanni. 1999. A tyrosinase peptide presented by HLA-B35 is recognized on a human melanoma by autologous cytotoxic T lymphocytes. Int. J. Cancer 83:755.[Medline]
  25. Beck, Y., L. Satz, Y. Takamiya, S. Nakayama, L. Ling, Y. Ishikawa, T. Nagao, H. Uchida, K. Tokunaga, C. Müller, T. Juji, M. Takiguchi. 1995. Polymorphism of human minor histocompatibility antigens: T cell recognition of human minor histocompatibility peptides presented by HLA-B35 subtype molecules. J. Exp. Med. 181:2037.[Abstract/Free Full Text]
  26. Smith, K. J., S. W. Reid, D. I. Stuart, A. J. McMichael, E. Y. Jones, J. I. Bell. 1996. An altered position of the {alpha}2 helix of MHC class I is revealed by the crystal structure of HLA-B*3501. Immunity 4:203.[Medline]
  27. Summers, C., F. Karst, A. D. Charles. 1993. Cloning, expression and characterisation of the cDNA encoding human hepatic squalene synthase, and its relationship to phytoene synthase. Gene 136:185.[Medline]
  28. Foulon, V., V. D. Antonenkov, K. Croes, E. Waelkens, G. P. Mannaerts, P. P. Van Veldhoven, M. Casteels. 1999. Purification, molecular cloning, and expression of 2-hydroxyphytanoyl-CoA lyase, a peroxisomal thiamine pyrophosphate-dependent enzyme that catalyses the carbon-carbon bond cleavage during alpha-oxidation of 3-methyl-branched fatty acids. Proc. Natl. Acad. Sci. USA 96:10039.[Abstract/Free Full Text]
  29. Rammensee, H.-G., J. Bachmann, S. Stevanovic. 1997. MHC Ligands and Peptide Motifs Springer, Berlin.
  30. Fields, B. A., B. Ober, E. L. Malchiodi, M. I. Lebedeva, B. C. Braden, X. Ysern, J. K. Kim, X. Shao, E. S. Ward, R. A. Mariuzza. 1995. Crystal structure of the V{alpha} domain of a T cell antigen receptor. Science 270:1821.[Abstract/Free Full Text]
  31. Garboczi, D. N., P. Ghosh, U. Utz, Q. R. Fan, W. E. Biddison, D. C. Wiley. 1996. Structure of the complex between human T-cell receptor, viral peptide and HLA-A2. Nature 384:134.[Medline]
  32. Heath, W. R., K. P. Kane, M. F. Mescher, L. A. Sherman. 1991. Alloreactive T cells discriminate among a diverse set of endogenous peptides. Proc. Natl. Acad. Sci. USA 88:5101.[Abstract/Free Full Text]
  33. Heath, W. R., L. A. Sherman. 1991. Cell-type-specific recognition of allogeneic cells by alloreactive cytotoxic T cells: a consequence of peptide-dependent allorecognition. Eur. J. Immunol. 21:153.[Medline]
  34. Cereb, N., S. Young Yang. 1996. Induction of microvariant-specific CTL lines reactive to a single amino acid mismatch in bulk cultures using a transfectant expressing a single HLA class I molecule. J. Immunol. 156:18.[Abstract]
  35. Panina-Bordignon, P., G. Corradin, E. Roosnek, A. Sette, A. Lanzavecchia. 1991. Recognition by class II alloreactive T cells of processed determinants from human serum proteins. Science 252:1548.[Abstract/Free Full Text]
  36. Moss, P. A. H., J. I. Bell. 1995. Sequence analysis of the human {alpha}ß T-cell receptor CDR3 region. Immunogenetics 42:10.[Medline]
  37. Tallquist, M. D., T. J. Yun, L. R. Pease. 1996. A single T cell receptor recognizes structurally distinct MHC/peptide complexes with high specificity. J. Exp. Med. 184:1017.[Abstract/Free Full Text]
  38. Udaka, K., T. J. Tsomides, P. Walden, N. Fukusen, H. N. Eisen. 1993. A ubiquitous protein is the source of naturally occurring peptides that are recognized by a CD8+ T-cell clone. Proc. Natl. Acad. Sci. USA 90:11272.[Abstract/Free Full Text]
  39. Burrows, S. R., T. B. Sculley, I. S. Misko, C. Schmidt, D. J. Moss. 1990. An Epstein-Barr virus-specific cytotoxic T cell epitope in EBV nuclear antigen 3 (EBNA 3). J. Exp. Med. 171:345.[Abstract/Free Full Text]



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