Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor.

Abstract

The existence of lymphocytes within melanoma deposits that, when isolated, are capable ofrecognizing specific tumor antigens on autologous and allogeneic melanomas in a major histocompatibility complex (MHC) restricted fashion provides strong evidence that an immune response to cancer exists in humans (1-8). The ability of these tumor-infiltrating lymphocytes (TILs) to mediate the regression ofcancer when adoptively transferred into patients with metastatic melanoma attests to the clinical importance of the antigens recognized (9, 10). Characterization of these antigens may thus be important for development of strategies for cancer immunotherapy. ) have characterized two genes coding for melanoma antigens, MAGE-1 and tyrosinase, by using T-cell clones established from the peripheral blood of patients who were repetitively immunized in vivo with mutagenized tumor cells or whose peripheral blood lymphocytes were sensitized by repetitive in vitro stimulation with tumor. The strategy used in the present study attempted to identify the genes coding for tumor antigens that were recognized by TILs from tumor-bearing patients in the absence of immunization to enhance the possibility that genes would be identified that coded for antigens involved in the natural immune response against the growing cancer. Anti-melanoma T cells appear to be enriched in TILs probably as a consequence of clonal expansion and accumulation at the tumor site in vivo (14). We have now cloned and sequenced a gene coding for a shared, commonly expressed melanoma antigen, restricted by HLA-A2.1. § MATERIALS AND METHODS Generation of Cytotoxic T Lymphocytes (CTLs) and Culture of Cell Lines. CTLs were generated from excised tumor specimens by culturing a suspension of cells with interleukin 2 (IL-2) (6000 units/ml) (Chiron) for

Dendritic cells transduced with gp100 gene by RGD fiber-mutant adenovirus vectors are highly efficacious in generating anti-B16BL6 melanoma immunity in mice

“…15,16,18 In recent years, the cloning of various MAAs interacting with the melanocyte differentiation and melanogenesis pathways has opened new possibilities for the development of active immunotherapy designed to cause the rejection of malignant melanoma. [19][20][21][22][23] gp100, an MAA, is consistently expressed in both murine and human melanomas, 27,40 and has been shown to be highly immunogenic and an important target for active-specific immunotherapy in humans. 41 However, mice injected with Ad 26 or recombinant vaccinia virus 27 encoding murine gp100, which was cloned from a B16 cDNA library, failed to produce any detectable T-cell responses or protective immunity against murine B16 melanoma.…”

“…These include gp100, MART-1/Melan-A, tyrosinase, and tyrosinase-related protein. [19][20][21][22][23] Among these defined melanoma-associated antigens (MAAs), which are nonmutated self-differentiation antigens associated with melanin synthesis in normal melanocytes, gp100 appears to be a promising target antigen. Several human MHC class I-restricted peptides from gp100 have been identified, and vaccination strategies using synthetic gp100-derived peptides in conjunction with chemical adjuvant or autologous DCs have been evaluated in early phase I/II clinical trials.…”

Recombinant E. coli efficiently delivers antigen and maturation signals to human dendritic cells: Presentation of MART1 to CD8+ T cells

“…As a consequence of the interaction with bacteria, human DCs undergo marked phenotypic and functional maturation. Furthermore, we show that fixed E. coli/ LLO expressing the well-characterised human melanoma antigen, MART1, 20,21 efficiently deliver the HLA-A2-restricted MART1 [27][28][29][30][31][32][33][34][35] epitope for processing and presentation on human MoDCs, suggesting the potential of this system as a novel strategy for human tumour immunotherapy.…”

Melanoma antigen expression in serial fine-needle aspiration samples in patients with metastatic malignant melanoma participating in immunotherapy clinical trials

“…It is recognized by cytotoxic T lymphocytes in the context of HLA-A*0201. 9,13,14,21 Anti-MART-1 murine monoclonal antibody, M2-7C10, was developed at the NCI 2 and shown to be superior to HMB-45 for the diagnosis of MMM in FNA samples. 10 In a recent large scale study performed in our institution, 84% of MMM lesions (80% of patients) were immunoreactive with anti-HMB-45 whereas 92% of MMM lesions (90% of patients) were immunoreactive with anti-MART-1.…”