H. Lee Moffitt Cancer Center & Research Institute

Tumor Biology for the Clinician: Melanoma Peptide Vaccines


Craig L. Slingluff, Jr, MD

Department of Surgery, University of Virginia Health Sciences Center


Introduction

Vaccines against viral disease are effective in preventing human infections. They are administered to prevent infections for which effective treatments are not available, especially when active infection is associated with high mortality. Human cancers, like some viral infections, are highly lethal, and existing treatments are largely inadequate. However, cancers differ from viral infections in numerous ways. Cancers have long latency periods, while viral infections typically have short incubation periods. Cancers in different individuals are not identical, whereas viruses in different individuals generally are identical or closely related. In most cases, cancer antigens are encoded by normal host genes, whereas viral antigens are foreign to the host.

To vaccinate successfully against melanoma, an antigen or a series of antigens expressed on melanomas from different individuals (shared antigens) is required. Protective or cytotoxic immune responses must be generated against these shared antigens, and side effects must be tolerable or absent. These requirements imply that the antigens must be immunogenic in humans and that patients being immunized must not be tolerant or anergic to them. Furthermore, the concern for a nontoxic regimen requires that cross-reactivity on normal tissues not be a significant side effect.

In murine tumor systems, a naive animal can be protected against subsequent tumor challenge by vaccination with irradiated syngeneic tumor cells,[1] and metastatic tumor deposits can be eradicated by adoptive transfer of immune lymphocytes.[2] Following these principles, patients with metastatic cancer have been treated with tumor vaccines and with adoptive transfer of tumor-specific cytotoxic T lymphocytes (CTLs). Complete regressions of metastatic tumor deposits have been observed in some of these patients.[3-6] These results with adoptive transfer of lymphocytes provide compelling evidence that antigens recognized by tumor-directed CTLs are relevant targets for immune rejection of human tumors. Results with immunotherapy in human trials, however, have not been as successful as those in murine tumor systems. Unlike murine subjects, human cancer patients are not immunologically naive subjects being vaccinated with autologous (syngeneic) tumor with the goal of preventing the establishment of a tumor challenge. Potential barriers to equivalent success in humans are the gradual growth of tumors over years, which could be tolerizing, and the difficulty of immunizing against antigens to which some degree of tolerance exists.

Experience With Tumor Vaccines

Despite these barriers, melanoma vaccines have been administered to patients for several decades in hopes of boosting immunity to the patient's melanoma, usually as an adjuvant to surgery. However, convincing therapeutic efficacy has not yet been demonstrated in prospective randomized trials.[7] Early efforts to evaluate responses to tumor vaccines focused on humoral responses. The result was the identification of many antigens that could be defined serologically on melanoma cells, including glycoproteins and gangliosides.[8-11] Adoptive therapies directed against these antigens have been disappointing, but there is renewed interest in humoral immune responses and in tumor vaccines directed against gangliosides,[12] with phase II studies suggesting some protective effect.

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Table 1. Cell-Based Tumor Vaccines


Whole cell

        allogeneic
                single cell line
                pooled cell lines

        autologous
                cytokine gene-transduced
                hapten-modified

Membrane preparation

        vaccinia oncolysate

Shed antigens

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Despite the long-term interest in humoral immune responses, the dominant thrust of current research in melanoma immunobiology has focused on defining antigens recognized by human T cells and on augmenting the cellular immune response to melanoma. The fact that T cells infiltrate many primary and metastatic melanoma deposits suggests the presence of a cellular immune response in vivo. However, the response appears to be inadequate, because tumors grow despite the infiltrate. One explanation for this may be the lack of costimulatory molecules (such as B7) on human melanoma cells.[13-15]

Evidence from adoptive cellular therapy trials[16] demonstrates that melanoma- specific CTLs expanded in vitro from metastatic tumor sites can eradicate metastatic melanoma deposits in some patients after adoptive transfer. Though not optimized, the response rates from this therapy suggest that tumor-specific CTLs can be an effective therapeutic agent against malignant disease. Ex vivo culture of human lymphocytes and subsequent reinfusion is labor-intensive, costly, and requires expertise that generally is not available at most medical centers. The stimulation of effective cellular immune responses in vivo would be preferable, which would take advantage of the host's immune system. The question then is how to achieve that goal with tumor vaccines.

Cell-Based Vaccines

The classic approach has been to immunize patients with viable, whole melanoma cells, either autologous or irradiated. This approach is based on the assumptions that human melanoma cells express antigens to which an immune response can be generated, that such a response protects against tumor recurrence, and that administration of melanoma cells after ex vivo preparation results in an appropriate immunologic milieu to induce protective immunity. Potential problems with cell-based melanoma vaccines include the following: The dosage of antigen is not quantified or reproducible. Intact tumor cells may be immunosuppressive due to lack of costimulation or secretion of certain cytokines. The risk of viral disease exists when allogeneic vaccines are used The use of autologous vaccines is limited to patients with measurable advanced disease. Specificity of response is difficult to evaluate.

Several cell-based tumor vaccines that have been or are being used are listed in Table 1. Autologous tumor cell vaccines have the advantage of expressing all the antigens relevant to autologous tumor rejection in the immunized patient. Allogeneic melanoma cells may not express all the antigens that are present on autologous tumor cells, but melanoma cells from different individuals who share major histocompatibility complex (MHC) molecules express shared antigens recognized by melanoma-specific CTLs.[17,18] Thus, allogeneic melanoma cells may express relevant antigens for stimulation of an effective immune response against autologous melanoma. One argument for using autologous cells in whole-cell vaccines is that the autologous tumor cells will present antigenic peptides in the context of self-MHC molecules; however, this model does not conform with some current beliefs. The majority of human melanoma cells fail to express the costimulatory molecules B7.1 and B7.2,[15] without which these melanoma cells may not be adequate antigen-presenting cells (APCs). Thus, it may be hypothesized that those peptides must be presented to the immune system by professional APCs that process cellular proteins derived from the immunizing melanoma cells. This is a hypothesis for which some evidence exists.[19] If accurate, the HLA type of the immunizing cell line may not have any significant impact on the immunogenicity in a host. Instead, the ability for immunogenic peptides to be presented on appropriate MHC molecules of potent antigen-presenting cells may be more important.

Cell-Free Tumor Vaccines

Vaccination with purified peptides or purified proteins becomes a possible consideration as the identity of some of the immunogenic MHC-associated peptides present on human melanoma cells, as well as the proteins from which they originate, become known. This prospect of cell-free tumor vaccines has several attractive features, including the ability to immunize with known, large quantities of specific antigens, the absence of risk of transferring infectious agents, the ability to evaluate immunologic responses in terms of known specific epitopes, and the opportunity to evaluate immunologic toxicity in terms of specific autoimmunity. In addition, any success with individual peptides could be multiplied by creating a cocktail of immunogenic peptides. Also, the reproducibility of this sort of cell-free vaccine and its anticipated safety would eventually lead to the possibility of immunizing patients at risk, such as those with family histories of melanoma and dysplastic nevi or those with multiple melanoma primaries.

During the past decade, tumor-specific CTLs have been generated in vitro from peripheral blood, draining lymph nodes, or metastatic tumor deposits of human cancer patients. These are mostly CD8+ cells whose target cell recognition is restricted by class I MHC molecules.[20,21] The existence of tumor-specific CTLs in cancer patients is evidence for an ongoing, though insufficient, immune response to these tumors.

Although tumor cells may "escape" from recognition by such CTLs by down- regulating expression of MHC[22-25] or accessory/costimulatory molecules,[14] by selection of antigen-loss variants,[26,27] or by alterations in antigen-processing pathways,[28] such observations do not explain the isolation of CTLs that can recognize and lyse the fresh autologous tumor from patients with growing cancers. Presumably, either this cellular immune response is too weak to be effective or specific tolerance or immune suppression has overcome specific immunity in vivo. To explain this phenomenon, it is necessary to obtain a better understanding of the antigens recognized by these CTLs.

Most antigens recognized by cytotoxic T cells are short peptides that are bound to class I MHC molecules, those peptides having been produced by the degradation of proteins that are produced inside the cell.[29] A major focus of tumor immunologists, therefore, is the identification of the subset of MHC-associated peptides that are selectively expressed on tumor cells and that act as epitopes for tumor-specific CTLs.

T-Cell-Defined Epitopes Expressed on Melanoma Cells

The first peptide epitope defined for a human tumor was identified by a cDNA library transfection strategy. It is restricted by HLA-A1 and encoded by a gene called MAGE-1.[30] CTLs from the same patient used to identify this epitope also recognize a distinct MAGE-1 peptide in association with HLA-C region antigen.[31] However, it has not been possible to demonstrate recognition of MAGE-1 epitopes restricted by these or other MHC molecules using tumor-directed CTLs from other patients.[6,32] MAGE-1 is expressed in approximately 20% to 40% of cancers of several different tissue types, including melanomas, breast cancers, non-small cell lung cancers, head and neck squamous cell cancers, and bladder cancers. Among normal tissues, it has been identified only in the testis.[32,33] Twelve members of the MAGE gene family have now been identified.[34] Although their expression patterns are all similar, their functions remain unknown. MAGE-3 also encodes a peptide epitope for HLA-A1-restricted CTLs (EVDPIGHLY),[35] and it has been possible to stimulate CTL responses to this peptide using lymphocytes from normal individuals.[36] This peptide is highly homologous to the HLA-A1-restricted peptide epitope from MAGE-1 (EADPTGHSY).

Other peptide epitopes defined on human melanoma cells are derived from molecules that are expressed in a tissue-specific, rather than tumor-specific, manner. CTLs from two patients were shown to recognize two different peptides derived from tyrosinase in association with HLA-A2.1 - YMNGTMSQV and MLLAVLYCL.[37,38] Tyrosinase is an enzyme involved in the enzymatic conversion of tyrosine as a step in melanin synthesis, and it is consequently expressed in normal melanocytes as well as most melanomas. Attempts to identify reactivity to these peptides in other HLA-A2+ patients have been largely unrewarding.[6,39] However, HLA-A24 has been shown to present tyrosinase-derived epitopes to tumor-specific CTLs,[40] and tyrosinase has been identified as the source protein for peptide epitopes for class II MHC-restricted, melanoma-specific CD4+ T-helper cells.[41]

Another tissue-specific protein, called gp100 or Pmel-17, has been identified as a source for HLA-A2.1-restricted CTL epitopes. The relevance of this protein has been independently identified by three different experimental approaches.[6,39,42] This protein is the target of the antibody HMB45, which is specific for melanoma and melanocytes and has been in clinical use by pathologists for several years.[43-46] Based on the correlation between HMB45 reactivity and recognition by a single tumor-infiltrating lymphocyte-derived CTL line, one group established that transfection of cells with the gene for gp100/Pmel-17 reconstituted the epitope recognized by this T cell.[42] A second study using this same T-cell line to screen transfected cDNA libraries identified not only the gp100/Pmel-17 gene as the source of the epitope, but also a minimal peptide from the sequence that reconstituted activity (LLDGTATLRL).[6,47] Independently, using tandem mass spectrometry to study naturally occurring HLA-A2.1-associated peptides on human melanoma, the nonamer peptide YLEPGPVTA was shown to reconstitute the epitope recognized by CTLs from all five melanoma patients tested.[39] This peptide is encoded by a portion of the gene for gp100 and is distinct from that recognized by the single CTL clone used in the other two studies. Gp100[46] and Pmel-17[48,49] are highly homologous but nonidentical sequences, and they probably represent allelic variants of the same protein. The expression of the protein is correlated with melanin biosynthesis, but its exact function is unclear.[49] Patients whose tumor-infiltrating lymphocytes (TILs) recognize gp100-derived epitopes had responses to adoptive therapy with those TILs.[47]

Another gene encoding an HLA-A2.1-restricted epitope on melanoma cells was identified independently by two different laboratories using cDNA library transfection methods.[47,50,51] The function of the encoded protein, called MART-1[47,50] or Melan-A,[51] is unknown. However, its tissue distribution is limited to cells of melanocytic origin, and it is expressed in both normal melanocytes and most human melanomas. The HLA-A2.1-restricted epitope(s) derived from this protein are recognized by CTLs from approximately 90% of patients studied.47

In addition to these peptide epitopes, others are being identified at a rapidly increasing pace. The extensive and growing experience with melanoma-specific CTLs has permitted identification of multiple CTL epitopes on melanoma cells, some of which are widely recognized and some of which may be recognizable with appropriate stimulation or vaccination. The two major types of antigens for melanoma-specific CTLs are (1) melanocyte differentiation antigens and (2) normal genes expressed only in tumor cells and in the testis. It has been observed anecdotally that spontaneous regression of melanoma or responses to immunotherapy may be accompanied by vitiligo (loss of melanin pigment), which may be associated with improved prognosis.[6,52,53] These findings are consistent with the earlier report of Anichini et al[54] that HLA-A2-restricted melanoma-specific CTL clones also recognize HLA-A2+ cultured melanocytes. The association between tumor immunity and autoimmunity is an area of intense interest.

It has been postulated that tumor immunity is weak or ineffective because the antigens are recognized by low-affinity T cells. With the recent identification of several CTL epitopes derived from normal gene products, it is evident that a component of tumor immunity is autoimmunity. For CTL recognition of these antigens to occur, tolerance to these antigens either must never occur or must be overcome in vivo or in vitro. In the case of MAGE antigens, expression in normal tissues is limited to the testis. The patient from whom MAGE-specific CTLs were generated is a woman.[55] In the absence of testicular tissue, tolerance to MAGE antigens may be absent. It also is possible that MAGE antigens are expressed in testicular cells lacking MHC expression, in which case tolerance also may be absent. In the case of tissue differentiation antigens, tyrosinase, Melan-A/MART-1, and gp100/Pmel-17, it is possible that tolerance has developed for these molecules expressed in melanocytes. Melanoma-specific CTLs recognize and lyse cultured melanocytes,[54] confirming the expression of some differentiation antigens by cultured melanocytes and suggesting that peripheral tolerance to these antigens can be overcome.

The concentration of tyrosinase peptides required to reconstitute epitopes for CTL recognition is much higher than that required for other defined peptide epitopes.[38] The affinity of these peptides for the HLA-A2 molecule is comparable to that of other high affinity peptides. Therefore, it has been postulated that the CTLs that recognize them are low-affinity CTLs. Their low affinity may permit them to escape intrathymic clonal deletion, while high- affinity CTLs against autoantigens are deleted.[38,56] Conversely, the peptide 946, from gp100 (YLEPGPVTA) reconstitutes an epitope with 50% maximal lysis at 1 to 10 pM (seven to eight orders of magnitude lower than the tyrosinase peptides and several orders of magnitude less than some viral peptides). This peptide has intermediate to low affinity for the HLA-A2 molecule; therefore, the CTLs that recognize it are believed to have high affinity for the epitope.[39] It is possible that clonal anergy is not induced on normal melanocytes if the epitope is presented in low quantity on normal cells or if appropriate costimulatory molecules or T-cell help is not available. To answer questions about host tolerance to the differentiation antigens and autoimmunity directed against them, it will be important to define more accurately the expression and recognition of these antigens in normal humans, as well as the expression of MHC molecules and costimulatory molecules on melanocytes. As peptide-based vaccine strategies are initiated, using these and other antigens, evaluations of cross-reactive immune responses against normal tissues will be critical. In melanoma patients experiencing regressions of metastatic tumor, vitiligo has been observed, but no clinically significant changes in other tissues of melanocytic origin have been observed.[6] However, if immune responses to solid tumors of other tissues also involve destruction of normal tissues sharing those antigens, the consequences may be more serious.

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Table 2. Investigational Aspects of Cell-Free Tumor Vaccines


Assess efficacy of individual peptides, whole proteins, or subunits

Evaluate dose-response relationships

Characterize antigen presentation pathways

Determine optimal adjuvants

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Table 3. Approaches for the Development of Tumor Vaccines From Peptide Epitopes for Melanoma-Specific Cytotoxic T Lymphocytes


Purified peptide plus adjuvant

Peptide conjugated to cytokine

Peptide-lipid conjugate

Vaccinia construct with minigene encoding peptide

Peptide-pulsed dendritic cells

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Development of Novel Tumor Vaccines for Melanoma

Many questions about how to vaccinate against melanoma (Table 2) need to be answered in terms of human immune systems and in the context of human MHC molecules. Though transgenic mouse models may function as reasonable models for some purposes, many of the questions about peptide vaccines need to be addressed in human clinical trials. A number of these trials are planned or are underway, and they include approaches ranging from using purified synthetic peptides plus adjuvant to introducing gene sequences encoding MHC-associated epitopes into APCs via viral vectors (Table 3). One aspect of these and related tumor vaccines under investigation is the choice of adjuvants.

While alum and other traditional adjuvants are effective at increasing immunogenicity of proteins and protein subunits for the purpose of generating serologic (antibody) responses, newer adjuvants are needed to assist in the generation of tumor-specific cell-mediated immunity. Alum is a poor choice for this purpose. While bacille Calmette-Guerin and other bacterial cell wall extracts have some efficacy at inducing cell-mediated immunity, they are fairly toxic. Attention currently is focusing on minimizing toxicity from bacterial cell wall products (eg, Detox by Ribi ImmunoChem Research, Inc, Hamilton, Mont) and studying alternate vaccine adjuvants including forms of oil emulsions related to incomplete Freund's adjuvant (eg, Montanide ISA-51 by Seppic, Inc., Paris, France) and saponins (eg, QS21 by Cambridge Biotech, Worcester, Mass).

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Table 4. Potential Causes of Vaccine Failure


Protective response not induced         Inadequate dose of vaccine
                                        Weak/inappropriate adjuvant
                                        CD4 helper arm not activated
                                        Peptide presentation incorrect
                                        Tolerance exists


Autoimmunity induced                    Vitiligo
                                        Ocular manifestations


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Table 5. Evaluation of Cellular Immune Responses to Tumor Vaccines


Delayed-type hypersensitivity response to peptide and to tumor cells

Cellular infiltrate at immunization site and distant tumor sites

Mixed lymphocyte tumor cell cultures/cytotoxic T lymphocytes 
generated in vitro

Precursor frequency analysis

Clinical tumor regression

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Phase I trials of peptide vaccine may need to be assessed in a different light than phase I trials of pharmacologic agents. In the latter, dose escalation is performed to determine a maximum tolerated dose. For peptide vaccines, it is difficult to imagine a major dose-related toxicity at any reasonable dose, and the optimal dose may not be the maximum tolerated dose. Immunologic effects and autoimmunity, if they occur, may appear at similar doses and may be much less dependent on dose than on the immunologic milieu in which the vaccine is administered. Evaluation of novel tumor vaccines depends on the need to evaluate those aspects of the immune response that are believed to be affected by the vaccine and to evaluate cross-reactivities and toxicities. Several potential causes of vaccine failure are listed in Table 4, and methods used in the evaluation of tumor vaccines are listed in Table 5.

If tumor vaccines derived from peptide epitopes for melanoma-specific CTLs can be used effectively to generate protective immunity against the immunizing peptide, then consideration of how to optimize the effective immune response will follow. Many mechanisms for tumor escape from immune recognition have been observed and proposed. An approach that almost certainly will be needed is the creation of cocktails of peptides or protein subunits, which would be administered with an effective adjuvant or a set of minigenes incorporated in a single viral vector.

Conclusions

During the past two years, the number of identified CTL epitopes for human melanoma has increased rapidly. Though mutated self-proteins might have been suspected as the predominant form of these epitopes, those that have been identified appear to be the result of the expression of normal genes. These genes include tissue differentiation antigens and latent genes expressed only in malignancy and in the testis. As we develop a better understanding of the relationship among tumor immunity, autoimmunity, and tolerance, then it also should be possible to identify failures of tumor immunity. The successful immunization against cancer may then become feasible, and methods for quantifying the status of the immune response - and for augmenting that response - may be developed. During the next decade, a better understanding of the immune response to human solid tumors is certain to emerge.

This work was supported by United States Public Health Service Grant CA57653.

References

1 Gross L. Intradermal immunization of C3H mice against a sarcoma that originated in an animal of the same line. Cancer Res. 1943;3:326-333.

2 Shu S, Chou T, Rosenberg SA. In vitro sensitization and expansion with viable tumor cells and interleukin 2 in the generation of specific therapeutic effector cells. J Immunol. 1986;136:3891-3898.

3 Rosenberg SA, Packard BS, Aebersold PM, et al. Use of tumor- infiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. N Engl J Med. 1988;319:1676-1680.

4 Berd D, Maguire HC Jr, McCue P, et al. Treatment of metastatic melanoma with an autologous tumor-cell vaccine: clinical and immunologic results in 64 patients. J Clin Oncol. 1990;8:1858-1867.

5 Mitchell MS, Habel W, Kan-Mitchell J, et al. Active specific immunotherapy of melanoma with allogeneic cell lysates. Rationale, results, and possible mechanisms of action. Ann N Y Acad Sci. 1993;690:153-166.

6 Kawakami Y, Eliyahu S, Delgado CH, et al. Identification of a human melanoma antigen recognized by tumor-infiltrating lymphocytes associated with in vivo tumor rejection. Proc Natl Acad Sci U S A. 1994;91:6458-6462.

7 Wallack MK, Sivanandham M, Balch CM, et al. A phase III randomized, double-blind, multiinstitutional trial of vaccinia melanoma oncolysate-active specific immunotherapy for patients with stage II melanoma. Cancer. 1995;75:34-42.

8 Bumol TF, Harper JR, Chee DO, et al. Antigenic expression of human melanoma cells in serum-free medium. Adv Exp Med Biol. 1984;172:455-470.

9 Cheresh DA, Honsik CJ, Staffileno LK, et al. Disialoganglioside GD3 on human melanoma serves as a relevant target antigen for monoclonal antibody-- mediated tumor cytolysis. Proc Natl Acad Sci U S A. 1985;82:5155-5159.

10 Yamaguchi H, Furukawa K, Fortunato SR, et al. Cell-surface antigens of melanoma recognized by human monoclonal antibodies. Proc Natl Acad Sci U S A. 1987;84:2416-2420.

11 Reisfeld RA. Antigen repertoire of human melanoma. Prog Clin Biol Res. 1988;256:343-360.

12 Livingston PO, Adluri S, Helling F, et al. Phase I trial of immunological adjuvant QS-21 with a GM2 ganglioside-keyhole limpet haemocyanin conjugate vaccine in patients with malignant melanoma. Vaccine. 1994;12:1275-1280.

13 Townsend SE, Allison JP. Tumor rejection after direct costimulation of CD8+ T cells by B7-transfected melanoma cells. Science. 1993;259:368-370.

14 Ramarathinam L, Castle M, Wu Y, et al. T cell costimulation by B7/BB1 induces CD8 T cell-dependent tumor rejection: an important role of B7/BB1 in the induction, recruitment, and effector function of antitumor T cells. J Exp Med. 1994;179:1205-1214.

15 Hersey P, Si Z, Smith MJ, et al. Expression of the co-stimulatory molecule B7 on melanoma cells. Int J Cancer. 1994;58:527-532.

16 Rosenberg SA, Yannelli JR, Yang JC, et al. Treatment of patients with metastatic melanoma with autologous tumor-infiltrating lymphocytes and interleukin 2. J Natl Cancer Inst. 1994;86:1159-1166.

17 Darrow TL, Slingluff CL Jr, Seigler HF. The role of HLA class I antigens in recognition of melanoma cells by tumor-specific cytotoxic T lymphocytes. Evidence for shared tumor antigens. J Immunol. 1989;142:3329-3335.

18 Slingluff CL Jr, Cox AL, Henderson RA, et al. Recognition of human melanoma cells by HLA-A2.1-restricted cytotoxic T lymphocytes is mediated by at least six shared peptide epitopes. J Immunol. 1993;150:2955-2963.

19 Huang AY, Golumbek P, Ahmadzadeh M, et al. Role of bone marrow-derived cells in presenting MHC class I-restricted tumor antigens. Science. 1994;264:961-965.

20 Chen Q, Hersey P. MHC-restricted responses of CD8+ and CD4+ T-cell clones from regional lymph nodes of melanoma patients. Int J Cancer. 1992;51:218-224.

21 Hom SS, Topalian SL, Simonis T, et al. Common expression of melanoma tumor-associated antigens recognized by human tumor infiltrating lymphocytes: analysis by human lymphocyte antigen restriction. J Immunother. 1991;10:153-164.

22 Blieden TM, McAdam AJ, Frelinger JG, et al. Mechanism of cytolytic T lymphocyte killing of a low class I-expressing tumor. J Immunol. 1991;147:1433-1438.

23 D'Alessandro G, Zardawi I, Grace J, et al. Immunohistological evaluation of MHC class I and II antigen expression on nevi and melanoma: relation to biology of melanoma. Pathology. 1987;19:339-346.

24 Levitsky HI, Lazenby A, Hayashi RJ, et al. In vivo priming of two distinct antitumor effector populations: the role of MHC class I expression. J Exp Med. 1994;179:1215-1224.

25 Vegh Z, Wang P, Vanky F, et al. Selectively down-regulated expression of major histocompatibility complex class I alleles in human solid tumors. Cancer Res. 1993;53(Suppl 10):2416-2420.

26 Knuth A, Wolfel T, Klehmann E, et al. Cytolytic T-cell clones against an autologous human melanoma: specificity study and definition of three antigens by immunoselection. Proc Natl Acad Sci U S A. 1989;86:2804-2808.

27 Seung S, Urban JL, Schreiber H. A tumor escape variant that has lost one major histocompatibility complex class I restriction element induces specific CD8+ T cells to an antigen that no longer serves as a target. J Exp Med. 1993;178:933-940.

28 Restifo NP, Esquivel F, Kawakami Y, et al. Identification of human cancers deficient in antigen processing. J Exp Med. 1993;177:265-272.

29 Townsend A, Bodmer H. Antigen recognition by class I-restricted T lymphocytes. Annu Rev Immunol. 1989;7:601-624.

30 Traversari C, van der Bruggen P, Luescher IF, et al. A nonapeptide encoded by human gene MAGE-1 is recognized on HLA-A1 by cytolytic T lymphocytes directed against tumor antigen MZ2-E. J Exp Med. 1992;176:1453-1457.

31 van der Bruggen P, Szikora JP, Boel P, et al. Autologous cytolytic T lymphocytes recognize a MAGE-1 nonapeptide on melanomas expressing HLA-Cw*1601. Eur J Immunol. 1994;24:2134-2140.

32 Zakut R, Topalian SL, Kawakami Y, et al. Differential expression of MAGE-1, -2, and -3 messenger RNA in transformed and normal human cell lines. Cancer Res. 1993;53:5-8.

33 van der Bruggen P, Traversari C, Chomez P, et al. A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science. 1991;254:1643-1647.

34 Boon T, Cerottini JC, Van den Eynde B, et al. Tumor antigens recognized by T lymphocytes. Annu Rev Immunol. 1994;12:337-365.

35 Gaugler B, Van den Eynde B, van der Bruggen P, et al. Human gene MAGE-3 codes for an antigen recognized on a melanoma by autologous cytolytic T lymphocytes. J Exp Med. 1994;179:921-930.

36 Celis E, Tsai V, Crimi C, et al. Induction of anti-tumor cytotoxic T lymphocytes in normal humans using primary cultures and synthetic peptide epitopes. Proc Natl Acad Sci U S A. 1994;91:2105-2109.

37 Brichard V, Van Pel A, Wolfel T, et al. The tyrosinase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med. 1993;178:489-495.

38 Wolfel T, Van Pel A, Brichard V, et al. Two tyrosinase nonapeptides recognized on HLA-A2 melanomas by autologous cytolytic T lymphocytes. Eur J Immunol. 1994;24:759-764.

39 Cox AL, Skipper J, Chen Y, et al. Identification of a peptide recognized by five melanoma-specific human cytotoxic T cell lines. Science. 1994;264:716-719.

40 Robbins PF, el-Gamil M, Kawakami Y, et al. Recognition of tyrosinase by tumor-infiltrating lymphocytes from a patient responding to immunotherapy. Cancer Res. 1994;54:3124-3126.

41 Topalian SL, Rivoltini L, Mancini M, et al. Human CD4+ T-cells specifically recognize a shared melanoma-associated antigen encoded by the tyrosinase gene. Proc Natl Acad Sci U S A. 1994;91:9461-9465.

42 Bakker AB, Schreurs MW, de Boer AJ, et al. Melanoma lineage-specific antigen gpl00 is recognized by melanoma-derived tumor-infiltrating lymphocytes. J Exp Med. 1994;179:1005-1009.

43 Smoller BR, Hsu A, Krueger J. HMB-45 monoclonal antibody recognizes an inducible and reversible melanocyte cytoplasmic protein. J Cutan Pathol. 1991;18:315-322.

44 Colombari R, Bonetti F, Zamboni G, et al. Distribution of melanoma- specific antibody (HMB-45) in benign and malignant melanocytic tumours: an immunohistochemical study on paraffin sections. Virchows Arch Pathol Anat Histopathol. 1988;413:17-24.

45 Adema GJ, de Boer AJ, van 't Hullenaar R, et al. Melanocyte lineage-specific antigens recognized by monoclonal antibodies NKI-beteb, HMB-50, HMB-45 are encoded by a single cDNA. Am J Pathol. 1993;143:1579-1585.

46 Vennegoor C, Hageman P, Van Nouhuijs H, et al. A monoclonal antibody specific for cells of the melanocyte lineage. Am J Pathol. 1988;130:179-192.

47 Kawakami Y, Eliyahu S, Sakaguchi K, et al. Identification of the immunodominant peptides of the MART-1 human melanoma antigen recognized by the majority of HLA-A2-restricted tumor infiltrating lymphocytes. J Exp Med. 1994;180:347-352.

48 Kwon BS, Halaban R, Kim GS, et al. A melanocyte-specific complementary DNA clone whose expression is inducible by melanotropin and isobutylmethylxanthine. Mol Biol Med. 1987;4:339-355.

49 Kwon BS, Chintamaneni C, Kozak CA, et al. A melanocyte-specific gene, Pmel 17, maps near the silver coat color locus on mouse chromosome 10 and is in a syntenic region on human chromosome 12. Proc Natl Acad Sci U S A. 1991;88:9228-9232.

50 Kawakami Y, Eliyahu S, Delgado CH, et al. Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor. Proc Natl Acad Sci U S A. 1994;91:3515-3519.

51 Coulie PG, Brichard V, Van Pel A, et al. A new gene coding for a differentiation antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med. 1994;180:35-42.

52 Bystryn JC, Rigel D, Friedman RJ, et al. Prognostic significance of hypopigmentation in malignant melanoma. Arch Dermatol. 1987;123:1053-1055.

53 Nordlund JJ, Kirkwood JM, Forget BM, et al. Vitiligo in patients with metastatic melanoma: a good prognostic sign. J Am Acad Dermatol. 1983;9:689-696.

54 Anichini A, Maccalli C, Mortarini R, et al. Melanoma cells and normal melanocytes share antigens recognized by HLA-A2-restricted cytotoxic T cell clones from melanoma patients. J Exp Med. 1993;177:989-998.

55 Boon T. Genes encoding tumor rejection antigens. Presented at Tumor Immunology Symposium; May 23, 1994; Charlottesville, Va.

56 Schwartz RH. Acquisition of immunologic self-tolerance. Cell. 1989;57:1073-1081.


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