REVIEWMelanoma antigens and their recognition by T cells
Giorgio Parmiani
Unit of Immunotherapy of Human Tumors, Istituto Nazionale Tumori, Milan, Italy(Received for publication on January 18, 2001)

Abstract. During the last 10 years many melanoma antigens recognized by T cells have been molecularly characterized. This review summarizes the main features of these antigens, including both classes I
and II HLA-restricted peptides, and describes their classi®cation into diverse groups according to thetissue distribution of the antigens. The different in vitro and in vivo immunogenicity of such antigens isthen discussed leading to the conclusion that Melan-A/MART-1 is the strongest among those tested
being frequently recognized by patients' T cells both in vitro and in vivo. However, no correlation was
found between T-cell response of melanoma patients to Melan-A/MART-1 and clinical response when
it was used for vaccination. Data are also presented that suggest, through an ex vivo analysis carriedout with tetramers staining of melanoma-speci®c T cells, that only in a limited number of advanced
patients does a speci®c immune response develop. This response, however, appears unable to effectively counteract metastatic melanoma growth. (Keio J Med 50 (2): 86±90, June 2001)
Key words: melanoma, antigens, T cells

verse phase high performance liquid chromatography(HPLC), screening by anti-melanoma speci®c T cells
and use of mass spectrometry to sequence the peptide
of interest.5
By using either of these approaches many new melanoma antigens and their epitopes recognized by T cells
in the context of class I HLA have been described.1,6More recently, techniques became available that can beused also to molecularly characterize class II HLArestricted melanoma epitopes recognized by CD4 T
helper lymphocytes, a ®nding which paves the way topossibly more effective vaccines for the immunotherapy
of melanoma patients. For a full list of these antigens
see Renkvist, et al.7

The molecular characterization of human tumor
antigens has opened a new era in cancer immunology
by allowing a more rapid translation of information
gained in basic research into clinical trials of immunotherapy.1 Melanoma is the tumor that was more frequently studied thanks to several clinical reports suggesting its antigenicity and to the availability of tumor
cell lines obtainable from melanoma patients and that
could be extensively used for in vitro immunologicalstudies.2 Melanoma antigens have been studied formany years at cellular level thanks to the ability toderive T cell clones from peripheral blood lymphocytes (PBL) or lymph node lymphocytes of melanoma
patients.3
Only molecular techniques, however, allowed cloning of genes encoding such protein antigens throughthe screening of cDNA expression tumor libraries bycytotoxic T lymphocytes (CTL) clones.4 An alternative
approach to the identi®cation of tumor (melanoma)
antigens was used which is also dependent on theavailability of patient's T cell clones but rests on abiochemical approach. This involves extraction of
antigenic peptides from major histocompatibility complex (MHC)/peptide complexes expressed on thecell surface of tumor cells, their fractionation by re-Classi®cation of Melanoma Antigens Recognized by
T Cells
According to their molecular nature and tissue distribution, melanoma antigens can be classi®ed as follows (Table 1):
Antigens predominantly expressed by tumor cells (orcancer/testis antigens)

Presented at the 1179th Meeting of The Keio Medical Society in Tokyo, October 3, 2000.
Reprint requests to: Dr. Giorgio Parmiani, Unit of Immunotherapy of Human Tumors, Istituto Nazionale per lo Studio e la Cura dei Tumori,
Via G. Venezian, 1-20133 Milano, Italy, e-mail: parmiani@istitutotumori.mi.it
86

Keio J Med 2001; 50 (2): 86±90
Table 1

87

Classi®cation of Human Melanoma Antigens Recognized by T Cells

Antigen

HLA-restriction

ExamplePredominantly expressed by tumor cells(cancer/testis)
Differentiation or melanocyte lineage-related
Broadley expressed
Unique antigens
Melanoma-speci®c antigens

Classes I and II*

MAGEClasses I and II*
Class IClasses I and IIClass I

gp100
p15
CDK4
TRP2-INT2

Tissue distribution
Melanoma and other tumors; testis;
placenta
Normal and neoplastic melanocytes
Normal and neoplastic tissues
Melanoma cells of a single tumor only
Shared among melanomas* The protein antigen contains different epitopes restricted either by class I or class II HLA. MAGE: melanoma antigen, CDK4: cyclinedependent kinase 4, TRP2: tyrosinase-related protein-2.

This group includes antigens represented by normal
proteins expressed by melanoma and other histologically different tumors but not by normal tissues with the
exception of testis. The prototype of this group of antigens is MAGE-1 (melanoma antigen-1), the ®rst humantumor antigen recognized by HLA-class I-restricted T
cells which has been molecularly characterized.4 In addition to the well known MAGE, BAGE and GAGE
families of antigens, recently a new cancer/testis antigen
has been described and called NY-ESO-1 owing to theplace where it was ®rst found (New York) and the histologically origin of the tumor (esophageous cancer)
from which the antigen was isolated by using patient's
serum antibodies and the Serex technique.8
The in vitro and in vivo immunogenicity of theseantigens is, in general, rather weak9 and no tumorin®ltrating lymphocytes (TIL) could ever be found inmelanoma patients directed against these families ofantigens. Generation of anti-MAGE-3 immunity bothin vitro and in vivo was, however, recently obtained byusing dendritic cells for its presentation to the immune
system.10
Differentiation antigens recognized by class I HLArestricted T cells
These antigens are shared between melanomas andnormal melanocytes and are thus lineage-related normal proteins mostly involved in the biosynthesis ofmelanin.11 These antigens are recognized by PBL,
which can be stimulated in vitro to develop antigenspeci®c CTL.12 Moreover, TIL recognizing such antigens in vitro in the presence of IL-2, were shown tocause clinical responses when re-infused in vivo inpatients whose tumor cells express the cognate antigen.13,14 Therefore, melanocyte differentiation antigens appear to be immunogenic at least in a fractionof patients with a hierarchy that indicates Melan-A/
MART1 as the most immunogenic one (see below).
Broadly expressed antigens

Other class I HLA-restricted antigens have beenreported consisting of normal proteins expressed widely
both on neoplastic and normal tissues, like p15 or
oncoproteins (e.g. Her2/neu) or oncosuppressive molecules (p53, p16) (see Table 1). Interestingly, recent
studies indicate that the protein domain of telomerase
can also provide epitopes expressed mostly by cancer
cells, including melanoma, and which are recognized byclass I HLA-restricted T cells.15 It is possible that many
normal protein epitopes expressed by normal tissues
are below the threshold level of T-cell recognition,
while their overexpression in tumor cells can trigger ananti-cancer response even by breaking a previously
established tolerance. This may be the case for the antigen PRAME which is detectable by reverse transcribed PCR in melanoma and normal tissues but is notrecognized by T cells in the latter thus becoming
operationally tumor-speci®c.16
Melanoma-speci®c, class I HLA-restricted unique andshared antigens
Important antigens are those expressed exclusively
by melanoma but not normal tissues. This group
includes two subgroups of antigens. Those generated bypoint mutation of normal proteins and expressed by
one single tumor only (unique antigens) like cyclinedependent kinase 4 (CDK4), b-catenin (see Table 2)that represent a biologically important feature of melanoma but, up to now, of impossible use in vaccination
trials owing to the dif®culty to prepare peptide-based
vaccines at the single patient level. The other subgroupencompasses at least two antigens which are shared
among melanomas without being found in normal tissues, including melanocytes. This kind of antigen, if
immunogenic enough, will represent the ideal vaccinethat should allow immunization of many patients without generating any toxic effect while sparing the normal
tissues from any harmful immunological reaction. We
found that HLA-A2 and HLA-A3-restricted epitopes
are contained in these antigens whose molecularnature was not completely elucidated.17,18 However, at

88Parmiani G: Melanoma antigens recognized by T cells

Table 2 Melanoma-Speci®c Unique and Shared Antigens Recognized by T Cells
Antigen

HLA-restriction

Mechanism of generation

UniqueCDK4b-catenin
Mum-1
Mum-2
CDC27
LDLR/FUTIn Vitro Immunogenicity of Melanoma Antigens

AntigenN. of positive reactions%

HLA-restriction

33
19
58
18

A2, B44
A2, A3, Cw8
A2
A1, A2
A1, A24
A2

N. of patients tested

Class IClass IClass IClass IClass IIClass II

Point mutation
Point mutation
Point mutation
Point mutation
Point mutation
Chromosome rearrangement

Tyrosinase
gp100
Melan-A/MART1
MAGE-3
MAGE-1GnT-VClass IClass I

Translation of intronic sequences
Translation of intronic sequences

Immunogenicity was evaluated by induction of antigen-speci®c CTLs
from PBL of melanoma patients after 1±5 weeks of stimulation withmelanoma cells or peptide-pulsed APC. GnT-V: acetylglucosaminyltransterase V.

SharedTRP2/INT
GnT-V

Table 3

CDK4: cycline-dependent kinase 4, TRP2: tyrosinase-related protein2, GnT-V: acetylglucosaminyl transterase V.

least two members of this subgroup of tumor-speci®cantigens have been molecularly characterized (Nacetylglucosaminyl transterase V (GnT-V) and (TRP(tyrosinase-related protein) -2-INT2)); it is of note thatin both cases the immunogenic epitopes were createdby the translation of retained intronic sequences.19,20
Class II HLA-restricted melanoma antigens
Immunogenicity of HLA class I-restricted melanoma
antigens has been found to be weak and only occasionally do patients develop a strong CTL reactivity after
immunization with molecularly de®ned vaccines.
Among the many possible reasons of such a phenomenon, the lack of activation of T helper cells may be
crucial inasmuch as CD4 T cells appear to be instrumental in eliciting a strong and sustained immune response against tumor antigens.21 Therefore, it is important to understand whether melanoma antigens
include epitopes that can activate a Th1 driven immune
reaction, which may ultimately result in melanoma cell
destruction. In the last few years several class II HLArestricted epitopes have been described. They fall into
the group of differentiation antigens (e.g. tyrosinase,
Melan-A/MART1)22,23 and cancer/testis antigens (e.g.
MAGE-3, NY-ESO-1)24±26 (see Table 1) but, interesting enough, several additional epitopes were found
to result from point mutations or chromosomal rearrangement of a variety of normal genes (see Table3).27,28 For a complete list and features of class II
HLA-restricted antigens see Renkvist, et al.7
It should be pointed out that, though the molecular
identi®cation of many melanoma antigens represents a
crucial progress in tumor immunology, from a practicalpoint of view the preparation of vaccines from a single
patient neoplasm (which should contain the whole
spectrum of potential antigens of that particular tumor)
still remains an unsolved problem. It is likely that truly

8/24
5/26
25/43
4/23
2/7
3/9

tumor-speci®c, immunogenic antigens are dif®cult to®nd and may include unique antigens that cannot becharacterized in a time compatible with their use inmetastatic melanoma patients in whom the tumor isprogressing. However, the use of heat-shock proteins(HSP) which, at least in the mouse system, are known
to bind the full repertoire of immunogenic tumor peptides, including the unique ones, and serve as chaperones for presentation of such peptides by dendritic cells
may allow the need of antigen identi®cation to be
bypassed.29 In fact, we have recently shown that HSPof 70 Kd puri®ed from human melanomas ef®ciently
present known melanoma antigens to patients' T
lymphocytes.30 This approach is feasible at individual
patient level provided enough tumor tissue is available
to purify HSP.
Recognition by T Cells in vitro and in vivo
Most of the antigens discussed above are recognizedby patient's T cells in vitro after repeated stimulations
and with a variable frequency in a population of melanoma patients. Results obtained by this approach,however, do not allow distinguishing between patients
whose immune system was already primed in vivo
against melanoma antigens owing to natural tumor
growth, from patients whose T cells were primed invitro by repeated antigenic stimulations. Nevertheless,the results of these experiments did demonstratethat patients often have T-cell precursors that canpotentially recognize melanoma antigens. The question
arises, however, which of the many known peptide
epitopes expressed by melanoma cells were the mostimmunogenic ones and, therefore, the best candidates
for a vaccination therapy.
Based on published data from several groups ofinvestigators, one can establish a hierarchy in terms ofin vitro immunogenicity of melanoma antigens used in
the form of peptides pulsed onto antigen-presenting
cells to stimulate patients' T cells (Table 3). From such

Keio J Med 2001; 50 (2): 86±90

an analysis, it appears that Melan-A/MART-1 27 ±35is the peptide epitope more frequently recognizedby patients' PBL after in vitro stimulation,31 possibly
thanks to a higher frequency of T cell precursors
against this antigen already present in normal donors32
as compared to other melanoma antigens. This conclusion was corroborated by subsequent studies of ex vivo
enumeration of melanoma antigen-speci®c T lymphocytes with tetramers,33 of immunization of HLA-A2transgenic mice given different HLA-A2-restricted
human melanoma peptides34 and, ®nally, of vaccination
trials showing that melanoma patients developed antiMelan-A/MART-1 CTL responses with a frequency
higher than that obtained by immunizing with gp100 or
tyrosinase peptides.35 It should be noted, however, thatCTL response elicited in patients after vaccination withmelanoma antigen peptides did not correlate with clinical responses.35±37
The availability of tetramers and the possibility to
study the in vivo distribution of tetramers plus T cells
and their function in vitro without the need to culture
these T cells, is shedding light on the complex interactions between melanoma growth, antigen expression, immune reactions and clinical outcome. In fact,
Anichini and co-workers have shown that 40% of untreated metastatic melanoma patients possess Melan-A/
MART-1-speci®c CD45RO memory T cells whichdisplay tumor cytotoxicity after two weeks of culture in
the presence of IL-2 while being ineffective within
metastatic lesions in vivo despite the presence of the
HLA-A2 restricting molecule and of the MART-1 asshown by immunohistochemistry.38 Moreover, a recent
re-evaluation of melanoma patients by using MART-1
tetramers to select melanoma antigen-speci®c T cells
has shown that the memory effector T cell phenotype(CD45RO, CCR7 ) can be found only in 25% of
metastatic melanoma patients, appears late in the progression of the disease, and is associated with a poor
prognosis.39 From these studies it can be concluded
that melanoma growth in vivo may activate a speci®c Tcell response against Melan-A/MART-1 but only aftera large tumor burden has been reached, this response
being then unable to ef®ciently control tumor growth.
This conclusion, however, is based on a few studies
and on the evaluation of the T-cell response against asingle melanoma antigen, namely Melan-A/MART-1,
which may not necessarily re¯ect the general behaviourof the kinetics of the immune response to the whole,unknown spectrum of melanoma antigens.
Concluding Remarks
Many melanoma antigens recognized by T cells in
the context of classes I or II MHC are now available
and several of these antigens are being used as vac-

89

cines, singly or in combination (polyepitopes vaccines).
However, we have still to ®nd out how to make melanoma antigens more immunogenic to patients' immune
system and to de®ne the kinetics of the T cell response
to such antigens during vaccination. The new technology of tetramers now allows such an analysis to be carried out at least for some antigens. A better knowledgeof the interplay between melanoma cell antigens andthe host's immune system may provide new insight tooptimize future protocols of vaccination.
Acknowledgements: The author's work is supported by the Italian
Associated for Cancer Research (Milan), by the Italian Ministry of
Health (Rome) and by the European Community (QLK3-199900064). The excellent editorial assistance of Ms Grazia Barp is gratefully acknowledged.

References
1. Kawakami Y, Rosenberg SA: T-cell recognition of self peptides
as tumor rejection antigens. Immunol Res 1996; 15: 179±190
2. Parmiani G, Anichini A, Fossati G: Cellular immune response
against autologous human malignant melanoma: are in vitro
studies providing a framework for a more effective immunotherapy? J Natl Cancer Inst 1990; 82: 361±370
3. Anichini A, Fossati G, Parmiani G: Clonal analysis of the cytolytic T-cell response to human tumors. Immunol Today 1987; 8:385±389
4. van der Bruggen P, Traversari C, Chomez P, Lurquin C, De
Plaen E, Van den Eynde B, Knuth A, Boon T: A gene encoding
an antigen recognized by cytolytic T lymphocytes on a humanmelanoma. Science 1991; 254: 1643±1647
5. Cox AL, Skipper J, Chen Y, Henderson RA, Darrow TL,
Shabanowitz J, Engelhard VH, Hunt DF, Slingluff CL Jr: Identi®cation of a peptide recognized by ®ve melanoma-speci®chuman cytotoxic T-cell lines. Science 1994; 264: 716±719
6. Castelli C, Rivoltini L, Andreola G, Carrabba M, Renkvist N,
Parmiani G: T-cell recognition of melanoma-associated antigens.
J Cell Physiol 2000; 182: 323±331
7. Renkvist N, Castelli C, Robbins PF, Parmiani G: A list of humantumor antigens recognized by T cells. Cancer Immunol Immunother 2001; 50: 3±15
È , Gure AO, Tsang S,
8. Chen YT, Scanlan MJ, Sahin U, TuÈreci O
Williamson B, Stockert E, Pfreundschuh M, Old LJ: A testicularantigen aberrantly expressed in human cancers detected byautologous antibody screening. Proc Natl Acad Sci USA 1997;
94: 1914±1918
9. Chaux P, Vantomme V, Coulie P, Boon T, van der Bruggen P:
Estimation of the frequencies of anti-MAGE-3 cytolytic Tlymphocyte precursors in blood from individuals without cancer.
Int J Cancer 1998; 77: 538±542
10. Schuler-Thurner B, Dieckmann D, Keikavoussi P, Bender A,
Maczek C, Jonuleit H, Roder C, Haendle I, Leisgang W, Dunbar
R, et al.: Mage-3 and in¯uenza-matrix peptide-speci®c cytotoxic
T cells are inducible in terminal stage HLA-A2.1 melanoma
patients by mature monocyte-derived dendritic cells. J Immunol
2000; 165: 3492±3496
11. Anichini A, Maccalli C, Mortarini R, Salvi S, Mazzocchi A,
Squarcina P, Herlyn M, Parmiani G: Melanoma cells and normalmelanocytes share antigens recognized by HLA-A2-restricted
cytotoxic T cell clones from melanoma patients. J Exp Med 1993;
177: 989±998

90
12. Rivoltini L, Kawakami Y, Sakaguchi K, Southwood S, Sette A,
Robbins PF, Marincola FM, Salgaller ML, Yannelli JR, AppellaE, et al.: Induction of tumor-reactive CTL from peripheral bloodand tumor-in®ltrating lymphocytes of melanoma patients by invitro stimulation with an immunodominant peptide of the humanmelanoma antigen MART-1. J Immunol 1995; 154: 2257±2265
13. Robbins PF, el-Gamil M, Kawakami Y, Stevens E, Yannelli JR,
Rosenberg SA: Recognition of tyrosinase by tumor-in®ltrating
lymphocytes from a patient responding to immunotherapy. Cancer Res 1994; 54: 3124±3126
14. Kawakami Y, Eliyahu S, Jennings C, Sakaguchi K, Kang X,
Southwood S, Robbins PF, Sette A, Appella E, Rosenberg SA:
Recognition of multiple epitopes in the human melanoma antigen gp100 by tumor-in®ltrating T lymphocytes associated with invivo tumor regression. J Immunol 1995; 154: 3961±3968
15. Vonderheide RH, Hahn WC, Schultze JL, Nadler LM: The
telomerase catalytic subunit is a widely expressed tumorassociated antigen recognized by cytotoxic T lymphocytes. Immunity 1999; 10: 673±679
16. Ikeda H, LetheÁ B, Lehmann F, Van Baren N, Baurain JF, deSmet C, Chambost H, Vitale M, Moretta A, Boon T, Coulie PG:
Characterization of an antigen that is recognized on a melanoma
showing partial HLA loss by CTL expressing an NK inhibitory
receptor. Immunity 1997; 6: 199±208
17. Anichini A, Mortarini R, Maccalli C, Squarcina P, Fleischhauer
K, Mascheroni L, Parmiani G: Cytotoxic T cells directed totumor antigens not expressed on normal melanocytes dominate
HLA-A2.1-restricted immune repertoire to melanoma. J Immunol 1996; 156: 208±217
18. Mazzocchi A, Storkus WJ, Traversari C, Tarsini P, Maeurer MJ,
Rivoltini L, Vegetti C, Belli F, Anichini A, Parmiani G, Castelli
C: Multiple melanoma-associated epitopes recognized by HLAA3-restricted CTLs and shared by melanomas but not melanocytes. J Immunol 1996; 157: 3030±3038
19. Guilloux Y, Lucas S, Brichard VG, Van Pel A, Viret C, De
Plaen E, Brasseur F, LetheÁ B, Jotereau F, Boon T: A peptide
recognized by human cytolytic T lymphocytes on HLA-A2 melanomas is encoded by an intron sequence of the N-acetylglucosaminyltransferase V gene. J Exp Med 1996; 183: 1173±1183
20. Lupetti R, Pisarra P, Verrecchia A, Farina C, Nicolini G, Anichini A, Bordignon C, Sensi M, Parmiani G, Traversari C:
Translation of a retained intron in tyrosinase-related protein(TRP) 2 mRNA generates a new cytotoxic T lymphocyte (CTL)de®ned and shared human melanoma antigen not expressed innormal cells of the melanocytic lineage. J Exp Med 1998; 188:1005±1016
21. Toes RE, Ossendorp F, Offringa R, Melief CJ: CD4 T cells andtheir role in antitumor immune responses. J Exp Med 1999; 189:
753±756
22. Topalian SL, Rivoltini L, Mancini M, Markus NR, Robbins PF,
Kawakami Y, Rosenberg SA: Human CD4 T cells speci®callyrecognize a shared melanoma-associated antigen encoded by thetyrosinase gene. Proc Natl Acad Sci USA 1994; 91: 9461±9465
23. Zarour HM, Kirkwood JM, Kierstead LS, Herr W, Brusic V,
Slingluff CL Jr, Sidney J, Sette A, Storkus WJ: Melan-A/MART1 (51±73) represents an immunogenic HLA-DR4-restricted
epitope recognized by melanoma-reactive CD4() T cells. Proc
Natl Acad Sci USA 2000; 97: 400±405
24. Chaux P, Vantomme V, Stroobant V, Thielemans K, Corthals J,
Luiten R, Eggermont AM, Boon T, van der Bruggen P: Identi®cation of MAGE-3 epitopes presented by HLA-DR molecules
to CD4() T lymphocytes. J Exp Med 1999; 189: 767±778
25. Manici S, Sturniolo T, Imro MA, Hammer J, Sinigaglia F,
Noppen C, Spagnoli G, Mazzi B, Bellone M, Dellabona P, et al.:Melanoma cells present a MAGE-3 epitope to CD4() cytotoxic
T cells in association with histocompatibility leukocyte antigen

Parmiani G: Melanoma antigens recognized by T cells
DR11. J Exp Med 1999; 189: 871±87626. Zeng G, Touloukian CE, Wang X, Restifo NP, Rosenberg SA,
Wang RF: Identi®cation of CD4 T cell epitopes from NY-ESO-1
presented by HLA-DR molecules. J Immunol 2000; 165: 1153±
1159
27. Pieper R, Christian RE, Gonzales MI, Nishimura MI, Gupta G,
Settlage RE, Shabanowitz J, Rosenberg SA, Hunt DF, Topalian
SL: Biochemical identi®cation of a mutated human melanoma
antigen recognized by CD4() T cells. J Exp Med 1999; 189:
757±766
28. Wang RF, Wang X, Atwood AC, Topalian SL, Rosenberg SA:
Cloning genes encoding MHC class II-restricted antigens:mutated CDC27 as a tumor antigen. Science 1999; 284: 1351±1354
29. Srivastava PK, Menoret A, Basu S, Binder RJ, McQuade KL:
Heat shock proteins come of age: Primitive functions acquire
new roles in an adaptive world. Immunity 1998; 8: 657±665
30. Castelli C, Ciupitu AM, Rini F, Rivoltini L, Mazzocchi A, Kiessling R, Parmiani G: Human heat shock protein 70 peptide
complexes speci®cally activate antimelanoma T cells. Cancer
Res 2001; 61: 222±227
31. Parmiani G: Future perspectives in speci®c immunotherapy ofmelanoma. Eur J Cancer 1998; 34(Suppl 3): S42±S47
32. Marincola FM, Rivoltini L, Salgaller ML, Player M, Rosenberg
SA: Differential anti-MART-1/Melan A CTL activity in peripheral blood of HLA-A2 melanoma patients in comparison to
healthy donors: evidence of in vivo priming by tumor cells. J
Immunother Emphasis Tumor Immunol 1996; 19: 266±277
33. Romero P, Dunbar PR, Valmori D, Pittet M, Ogg GS, Rimoldi
D, Chen JL, LieÂnard D, Cerottini JC, Cerundolo V: Ex vivo
staining of metastatic lymph nodes by class I major histocompatibility complex tetramers reveals high numbers of antigen-experienced tumor-speci®c cytolytic T lymphocytes. J Exp
Med 1998; 188: 1641±1650
34. Mateo L, Gardner J, Chen Q, Schmidt C, Down M, Elliott SL,
Pye SJ, Firat H, Lemonnier FA, Cebon J, Suhrbier A: An HLAA2 polyepitope vaccine for melanoma immunotherapy. J Immunol 1999; 163: 4058±4063
35. JaÈger E, Bernhard H, Romero P, Ringhoffer M, Arand M,
Karbach J, Ilsemann C, Hagedorn M, Knuth A: Generation ofcytotoxic T-cell responses with synthetic melanoma-associatedpeptides in vivo: implications for tumor vaccines with melanomaassociated antigens. Int J Cancer 1996; 66: 162±169
36. Cormier JN, Salgaller ML, Prevette T, Barracchini KC, Rivoltini
L, Restifo NP, Rosenberg SA, Marincola FM: Enhancementof cellular immunity in melanoma patients immunized with apeptide from MART-1 Melan-A. Cancer J Sci Am 1997; 3: 37±44
37. Marchand M, van Baren N, Weynants P, Brichard V, DreÂno B,
Tessier MH, Rankin E, Parmiani G, Arienti F, Humblet Y, et al.:Tumor regressions observed in patients with metastatic melanoma treated with an antigenic peptide encoded by gene
MAGE-3 and presented by HLA-A1. Int J Cancer 1999; 80:219±230
38. Anichini A, Molla A, Mortarini R, Tragni G, Bersani I, Di
Nicola M, Gianni AM, Pilotti S, Dunbar R, Cerundolo V,
Parmiani G: An expanded peripheral T cell population to acytotoxic T lymphocyte (CTL)-de®ned, melanocyte-speci®c antigen in metastatic melanoma patients impacts on generation of
peptide-speci®c CTLs but does not overcome tumor escape from
immune surveillance in metastatic lesions. J Exp Med 1999; 190:
651±667
39. Dunbar PR, Smith CL, Chao D, Salio M, Shepherd D, Mirza F,
Lipp M, Lanzavecchia A, Sallusto F, Evans A, et al.: A shift in
the phenotype of Melan-A-speci®c CTL identi®es melanoma
patients with an active tumor-speci®c immune response. J
Immunol 2000; 165: 6644±6652