title: Heterogeneous expression of immunotherapy candidate proteins gp100, MART-1, and tyrosinase in human melanoma cell lines and in human melanocytic lesions.
abstract: In recent years, it has become evident that T cells can recognize peptides of melanocytic lineage antigens such as gp100, MART-1, and tyrosinase at the tumor cell surface and can subsequently destroy these cells. It is thus feasible to develop immunotherapeutic approaches based on the melanocytic marker profiles of melanoma cells. One of the predictors of the success rate of such a treatment is the extent of positive (target) tumor cells within the lesions of the patient. First, we investigated the presence of these three proteins in 18 human melanoma cell lines using RT-PCR and immunohistochemistry. In 11 cell lines, mRNA and protein of all three markers could be detected; in one cell line, only two markers were present, and six melanoma cell lines showed no evidence for these markers. Secondly, we stained frozen sections of 105 human melanocytic lesions, 13 common nevocellular nevi, 13 atypical nevi, 13 early primary melanomas (Breslow < 1.5 mm), 25 advanced primary melanomas (aPM; Breslow > or =1.5 mm), and 41 melanoma metastases (MM) with antibodies against glycoprotein 100, melanoma antigen recognized by T cells, and tyrosinase. In addition, we used the 3,4-dihydroxy-L-phenylalanine reaction to detect tyrosinase enzyme activity as a confirmation of the tyrosinase immunohistochemical results in a subset of the lesions. In the benign lesions, glycoprotein 100 was more prominently expressed in epidermal melanocytes, whereas melanoma antigen recognized by T cells was encountered in all or nearly all dermal melanocytes in all nevocellular nevi and atypical nevus lesions. Tyrosinase was found in a lower percentage of melanocytes, both in the epidermis and in the dermis within these lesions. With regard to heterogeneity of staining within the malignant lesions, we found that 54% (early primary melanomas), 48% (aPMs), and 56% (MM) of the lesions stained within the same staining category for all three proteins studied. Approximately 17% of the aPM and MM lesions did not show positive tumor cells for any of the three proteins. We conclude that a subgroup of patients with high expression should be selected for immunotherapeutic treatment approaches based on the presence of these proteins.
authors.affiliation: Department of Pathology, University Hospital, Nijmegen, The Netherlands. t.devries@pathol.azn.nl
authors.forename: T J
authors.initials: TJ
authors.lastname: de Vries
authors.forename: A
authors.initials: A
authors.lastname: Fourkour
authors.forename: T
authors.initials: T
authors.lastname: Wobbes
authors.forename: G
authors.initials: G
authors.lastname: Verkroost
authors.forename: D J
authors.initials: DJ
authors.lastname: Ruiter
authors.forename: G N
authors.initials: GN
authors.lastname: van Muijen
mesh_terms: D000951:Antigens, Neoplasm; D004295:Dihydroxyphenylalanine / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D058965:MART-1 Antigen; D008545:Melanoma / Q000175:diagnosis / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000378:metabolism*; D011379:Prognosis; D012333:RNA, Messenger / Q000032:analysis; D012878:Skin Neoplasms / Q000175:diagnosis / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D058951:gp100 Melanoma Antigen
chemical_list: D000951:Antigens, Neoplasm; D058965:MART-1 Antigen; C548667:MLANA protein, human; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C548637:PMEL protein, human; D012333:RNA, Messenger; D058951:gp100 Melanoma Antigen; D004295:Dihydroxyphenylalanine; D014442:Monophenol Monooxygenase