Serological analysis of Melan-A(MART-1), a melanocyte-specific protein homogeneously expressed in human melanomas.

Chen, Yao‐Tseng; Stockert, Elisabeth; Jungbluth, Achim A.; Tsang, Solam; Coplan, Keren; Scanlan, Matthew J.; Old, Lloyd J.

doi:10.1073/pnas.93.12.5915

Cited by 194 publications

(155 citation statements)

References 22 publications

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“…The screening strategy used to detect MAb CT7-33 was similar to the approach we used to generate MAbs to other tumor antigens, e.g., MAb MA454 to MAGE-A9, MAb ES121 to NY-ESO-1, MAb A103 to Melan-A/MART-1 and MAb T311 to tyrosinase. 12,13,[17][18][19][20][21] Our screening strategy also employed formalin-fixed, paraffin-embedded tissues in the early immunohistochemical selection of the newly generated clones to ensure that the new reagent could be applied to standard archival material. Testis served as an immunohistochemical test tissue and proved, as in our previous studies, to be a suitable standard for selecting reactive clones.…”

Section: Discussionmentioning

confidence: 99%

CT7 (MAGE‐C1) antigen expression in normal and neoplastic tissues

Jungbluth

Chen

Busam

et al. 2002

Intl Journal of Cancer

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CT7 (MAGE-C1) is a member of the cancer testis (CT)antigen family. The present study describes the generation of CT7-33, a monoclonal antibody (MAb) to CT7, and the preliminary protein expression analysis of CT7 in normal tissues and in a limited number of neoplastic lesions. CT7-33 was effective in frozen as well as formalin-fixed, paraffin-embedded tissues, and immunohistochemistry/reverse transcriptase polymerase chain reaction (RT-PCR) co-typing demonstrated antibody specificity. CT7-33 immunoreactivity in normal adult tissues is restricted to testicular germ cells. In neoplastic lesions, CT7-33 immunostaining is confined to tumor cells, and the frequency of CT7 protein expression mostly parallels previous mRNA analyses. Whereas colorectal and renal cell carcinomas, as well as sarcomas, exhibit poor or no CT7-33 staining, carcinomas of the mammary gland and ovary, nonsmall cell lung carcinoma and metastatic melanomas exhibit a high incidence of CT7 protein expression. However, as seen in previous analyses of other CT antigens, the expression pattern is mostly heterogeneous, and tumors with more than 50% of tumor staining are only infrequently encountered. In summary, our study Key words: CT7; MAGE-C1; heterogeneous antigen expression; monoclonal antibody CT7-33Cancer testis (CT) antigens are expressed in a variety of malignant neoplasms but not in normal adult tissues except for germ cells of the testis. 1,2 Due to their expression pattern, CT antigens appear to be ideal targets for immunotherapeutic strategies against malignant neoplasms. Since the isolation of the prototype CT gene family, MAGE, the number of CT antigens has steadily increased. 3 Whereas autologous T-cell epitope cloning was used for the initial identification of CT antigens such as MAGE and BAGE, 4,5 others have been isolated by SEREX (serological analysis of recombinant cDNA expression libraries) analysis. SEREX employs the immunologic screening of cDNA expression libraries from human tumors with autologous patient sera. 6 -9 By using the SEREX approach, we recently identified CT7, 10 a new member of the CT family. The same gene was isolated independently by Lucas et al. 11 using representational difference analysis (RDA) and termed MAGE-C1. A preliminary RT-PCR analysis revealed the presence of CT7 (MAGE-C1) in normal testis and in a variety of neoplasms, such as melanoma, and tumors of the urinary bladder, breast and lung. To assess CT7 as a potential target for immunotherapy, analysis of its actual antigen expression is essential. The present study describes the generation of the monoclonal antibody (MAb) CT7-33 to CT7 (MAGE-C1) and an immunohistochemical analysis of CT7 (MAGE-C1) expression in normal and tumorous tissues. MATERIAL AND METHODS Production of recombinant CT7 proteinA truncated CT7 product was chosen for the expression in E. coli. This truncated form encoded 350 of 1,124 amino acids in the CT7 antigen, namely positions 552-901. The purpose of this truncation was to exclude the C-terminus, which shares significant homolo...

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Section: Discussionmentioning

confidence: 99%

CT7 (MAGE‐C1) antigen expression in normal and neoplastic tissues

Jungbluth

Chen

Busam

et al. 2002

Intl Journal of Cancer

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“…We (De Vries et al, 1997) and others (Chen et al, 1995(Chen et al, , 1996 have previously reported on the specificity of the antibodies used. Consecutive sections of all melanocytic lesions were immunohistochemically stained, using the above-mentioned antibodies as primary antibodies.…”

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confidence: 91%

“…MART-I (Chen et al 1996). tyrosinase (Chen et al 1995) and TRP-1 (Chen et al 1995) have recentlx been described.…”

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confidence: 99%

“…Antibodies and immunohistochemistry NKI-beteb (Monosan/Sanbio, Uden, The Netherlands) and HMB-45 (Dako, Glostrup, Denmark) were used as antibodies against gplOO (Adema et al, 1993), A103 (Chen et al, 1996) was used as antibody against MART-1 (Novocastra, Newcastle, UK), T311 (Chen et al, 1995) was used as antibody against tyrosinase and TA099 (Chen et al, 1995) was the antibody against TRP-1. We (De Vries et al, 1997) and others (Chen et al, 1995(Chen et al, , 1996 have previously reported on the specificity of the antibodies used.…”

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confidence: 99%

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High expression of immunotherapy candidate proteins gp100, MART-1, tyrosinase and TRP-1 in uveal melanoma

Vries

Trancikova²,

Ruiter³

et al. 1998

Br J Cancer

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Summary In the treatment of cutaneous melanoma, provisional therapeutic strategies have been designed to combat tumour load using T cells that are sensitized with peptides derived from melanoma autoantigens, such as glycoprotein 100 (gp100), melanoma antigen recognized by T cells 1 (MART-1 or MelanA), tyrosinase and tyrosinase-related protein 1 (TRP-1). We recently found that gpl 00, MART-1 and tyrosinase are heterogeneously expressed in human cutaneous melanoma (De Vries et al (1997) Monoclonal antibodies against gplOO has-e successfullv been implemented in diagnostic pathology of cutaneous and uveal melanoma (Carrel and Rimoldi. 1993: Ruiter andBrocker. 1993).Antibodies against three other melanoma antigens. MART-I (Chen et al. 1996). tyrosinase (Chen et al. 1995) and TRP-1 (Chen et al.1995) have recentlx been described. The recent discoxerx that peptides derived from gplOO (Bakker et al. 1994 (Tobal et al. 1993: Foss et al. 1995.For both those who design immunotherapy protocols and those w ho perform RT-PCRs based on the presence of melanomaspecific mRNA in patients blood, it is important to knoxx the content of these antigens in primary tumours. Recentlv. we studied the presence of gp 100. MART-1 and tyrosinase in cutaneous melanocvtic lesions. We found that approximately 20% of the

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“…On thawing, the numbers of tumor cells were counted as well as the number of lymphocytes, to provide an index of the extent of tumor infiltration, expressed as a percentage of the total cell count accounted for by tumor cells. Expression of melan-A was examined in TILN by RT-PCR (2), and protein expression confirmed in one sample by Western blotting using the primary Ab A103 (Novocastra, Newcastle, U.K.) (25). When tumor samples were available from patients, these were examined by immunohistochemistry, including staining for melan-A with the primary Ab A103 (Novocastra) (25).…”

Section: Subjects and Samplesmentioning

confidence: 99%

A Shift in the Phenotype of Melan-A-Specific CTL Identifies Melanoma Patients with an Active Tumor-Specific Immune Response

Dunbar¹,

Smith²,

Chao

et al. 2000

The Journal of Immunology

123

117

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In a significant proportion of melanoma patients, CTL specific for the melan-A26/7–35 epitope can be detected in peripheral blood using HLA-A2/peptide tetramers. However, the functional capacity of these CTL has been controversial, since although they prove to be effective killers after in vitro expansion, in some patients they have blunted activation responses ex vivo. We used phenotypic markers to characterize melan-A tetramer+ cells in both normal individuals and melanoma patients, and correlated these markers with ex vivo assays of CTL function. Melanoma patients with detectable melan-A tetramer+ cells in peripheral blood fell into two groups. Seven of thirteen patients had a CCR7+ CD45R0− CD45RA+ phenotype, the same as that found in some healthy controls, and this phenotype was associated with a lack of response to melan-A peptide ex vivo. In the remaining six patients, melan-A tetramer+ cells were shifted toward a CCR7− CD45R0+ CD45RA− phenotype, and responses to melan-A peptide could be readily demonstrated ex vivo. When lymph nodes infiltrated by melan-A-expressing melanoma cells were examined, a similar dichotomy emerged. These findings demonstrate that activation of melan-A-specific CTL occurs in only some patients with malignant melanoma, and that only patients with such active immune responses are capable of responding to Ag in ex vivo assays.

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Serological analysis of Melan-A(MART-1), a melanocyte-specific protein homogeneously expressed in human melanomas.

Cited by 194 publications

References 22 publications

CT7 (MAGE‐C1) antigen expression in normal and neoplastic tissues

CT7 (MAGE‐C1) antigen expression in normal and neoplastic tissues

High expression of immunotherapy candidate proteins gp100, MART-1, tyrosinase and TRP-1 in uveal melanoma

A Shift in the Phenotype of Melan-A-Specific CTL Identifies Melanoma Patients with an Active Tumor-Specific Immune Response

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