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MELOE-1 is a new antigen overexpressed in melanomas and involved in adoptive T cell transfer efficiency.

Godet Y, Moreau-Aubry A, Guilloux Y, Vignard V, Khammari A, Dreno B, Jotereau F, Labarriere N - J. Exp. Med. (2008)

Bottom Line: Remarkably, the structure of meloe was unusual, with multiple short open reading frames (ORFs).The peptide recognized by the CTL clone was encoded by one of these ORFs, called MELOE-1.Using a specific HLA-A2/peptide tetramer, we showed a correlation between the infusion of TILs containing MELOE-1-specific T cells and relapse prevention in HLA-A2 patients.

View Article: PubMed Central - PubMed

Affiliation: Institut National de Santé et de Recherche Médicale, Unité Mixte de Recherche 892, 44093 Nantes, France.

ABSTRACT
A cytotoxic T lymphocyte (CTL) clone was derived from a tumor-infiltrating lymphocyte (TIL) population infused to a melanoma patient who remained relapse free for 10 yr after this adoptive transfer. This clone recognized all melanoma cell lines tested and, to a lower extent, melanocytes, in the context of human histocompatibility leukocyte antigen A2 (HLA-A2), but it did not recognize other tumor cell types. The gene coding for the antigen recognized by this clone was identified by the screening of a melanoma complementary DNA expression library. This antigen is overexpressed in melanomas, compared with other cancer cell lines and healthy tissues, and was thus called melanoma-overexpressed antigen (meloe). Remarkably, the structure of meloe was unusual, with multiple short open reading frames (ORFs). The peptide recognized by the CTL clone was encoded by one of these ORFs, called MELOE-1. Using a specific HLA-A2/peptide tetramer, we showed a correlation between the infusion of TILs containing MELOE-1-specific T cells and relapse prevention in HLA-A2 patients. Indeed, 5 out of 9 patients who did not relapse were infused with TILs that contained MELOE-1-specific T cells, whereas 0 out of the 21 patients who relapsed was infused with such TIL-containing lymphocytes. Overall, our results suggest that this new antigen is involved in immunosurveillance and, thus, represents an attractive target for immunotherapy protocols of melanoma.

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Characterization of the cDNA coding for the recognized antigen. (A) M170.48 TNF responses to COS-7 cells (E/T ratio = 1:3) transfected with the indicated plasmids. The T cell clone was added 2 d after the transfection, and the CTL clone reactivity was assessed by a TNF release assay. (B) Comparison of the nucleotide sequences of meloe and BC008026 cDNAs and the localization of this sequence on the HDAC-4 gene. The indicated nucleotides correspond to SNPs between the meloe sequence isolated from M134 and A498 tumor cell lines and the meloe sequence isolated from the M117 and SW480 cell lines, and the BC008026 cDNA sequence.
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fig2: Characterization of the cDNA coding for the recognized antigen. (A) M170.48 TNF responses to COS-7 cells (E/T ratio = 1:3) transfected with the indicated plasmids. The T cell clone was added 2 d after the transfection, and the CTL clone reactivity was assessed by a TNF release assay. (B) Comparison of the nucleotide sequences of meloe and BC008026 cDNAs and the localization of this sequence on the HDAC-4 gene. The indicated nucleotides correspond to SNPs between the meloe sequence isolated from M134 and A498 tumor cell lines and the meloe sequence isolated from the M117 and SW480 cell lines, and the BC008026 cDNA sequence.

Mentions: We screened a cDNA library derived from the M134 melanoma cell line (20) in COS-7 cells cotransfected with pHLA-A*0201 to characterize the antigen recognized by the M170.48 T cell clone. Among 800 pools of 100 pcDNA tested, one plasmid pool proved positive in this test, and the individual plasmid coding for the recognized antigen was recovered from it after a cloning step. This insert, namely meloe, spanning 2,128 bp, was sequenced and was found to contain a poly(A) tail and to be similar to the clone BC008026 isolated by the National Institutes of Health (NIH) Mammalian Gene Collection consortium (21). After expression vector cloning, cotransfection of BC008026 cDNA with HLA A*0201 into COS-7 cells also induced the recognition by the M170.48 T cell clone (Fig. 2 A), although these two sequences differed by four single nucleotide polymorphisms (SNPs; Fig. 2 B). To control the impact of these SNPs on the recognition of the M170.48 T cell clone, we sequenced the cDNA isolated from recognized (M134 and M117) and nonrecognized cell lines (A498 and SW480), and showed that this polymorphism did not affect tumor cell line recognition. The meloe sequence analysis showed a perfect colinearity with the genomic DNA, obtained by comparison with the sequence of the human genome released by Celera (22), which indicated absence of splicing. Finally, this sequence was found to be located in the third intron of the histone deacetylase (HDAC) 4 gene (Ensembl accession no. ENSG00000068024), on chromosome 2, in anti-sense orientation compared with the sequence of the HDAC-4 gene.


MELOE-1 is a new antigen overexpressed in melanomas and involved in adoptive T cell transfer efficiency.

Godet Y, Moreau-Aubry A, Guilloux Y, Vignard V, Khammari A, Dreno B, Jotereau F, Labarriere N - J. Exp. Med. (2008)

Characterization of the cDNA coding for the recognized antigen. (A) M170.48 TNF responses to COS-7 cells (E/T ratio = 1:3) transfected with the indicated plasmids. The T cell clone was added 2 d after the transfection, and the CTL clone reactivity was assessed by a TNF release assay. (B) Comparison of the nucleotide sequences of meloe and BC008026 cDNAs and the localization of this sequence on the HDAC-4 gene. The indicated nucleotides correspond to SNPs between the meloe sequence isolated from M134 and A498 tumor cell lines and the meloe sequence isolated from the M117 and SW480 cell lines, and the BC008026 cDNA sequence.
© Copyright Policy
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2571940&req=5

fig2: Characterization of the cDNA coding for the recognized antigen. (A) M170.48 TNF responses to COS-7 cells (E/T ratio = 1:3) transfected with the indicated plasmids. The T cell clone was added 2 d after the transfection, and the CTL clone reactivity was assessed by a TNF release assay. (B) Comparison of the nucleotide sequences of meloe and BC008026 cDNAs and the localization of this sequence on the HDAC-4 gene. The indicated nucleotides correspond to SNPs between the meloe sequence isolated from M134 and A498 tumor cell lines and the meloe sequence isolated from the M117 and SW480 cell lines, and the BC008026 cDNA sequence.
Mentions: We screened a cDNA library derived from the M134 melanoma cell line (20) in COS-7 cells cotransfected with pHLA-A*0201 to characterize the antigen recognized by the M170.48 T cell clone. Among 800 pools of 100 pcDNA tested, one plasmid pool proved positive in this test, and the individual plasmid coding for the recognized antigen was recovered from it after a cloning step. This insert, namely meloe, spanning 2,128 bp, was sequenced and was found to contain a poly(A) tail and to be similar to the clone BC008026 isolated by the National Institutes of Health (NIH) Mammalian Gene Collection consortium (21). After expression vector cloning, cotransfection of BC008026 cDNA with HLA A*0201 into COS-7 cells also induced the recognition by the M170.48 T cell clone (Fig. 2 A), although these two sequences differed by four single nucleotide polymorphisms (SNPs; Fig. 2 B). To control the impact of these SNPs on the recognition of the M170.48 T cell clone, we sequenced the cDNA isolated from recognized (M134 and M117) and nonrecognized cell lines (A498 and SW480), and showed that this polymorphism did not affect tumor cell line recognition. The meloe sequence analysis showed a perfect colinearity with the genomic DNA, obtained by comparison with the sequence of the human genome released by Celera (22), which indicated absence of splicing. Finally, this sequence was found to be located in the third intron of the histone deacetylase (HDAC) 4 gene (Ensembl accession no. ENSG00000068024), on chromosome 2, in anti-sense orientation compared with the sequence of the HDAC-4 gene.

Bottom Line: Remarkably, the structure of meloe was unusual, with multiple short open reading frames (ORFs).The peptide recognized by the CTL clone was encoded by one of these ORFs, called MELOE-1.Using a specific HLA-A2/peptide tetramer, we showed a correlation between the infusion of TILs containing MELOE-1-specific T cells and relapse prevention in HLA-A2 patients.

View Article: PubMed Central - PubMed

Affiliation: Institut National de Santé et de Recherche Médicale, Unité Mixte de Recherche 892, 44093 Nantes, France.

ABSTRACT
A cytotoxic T lymphocyte (CTL) clone was derived from a tumor-infiltrating lymphocyte (TIL) population infused to a melanoma patient who remained relapse free for 10 yr after this adoptive transfer. This clone recognized all melanoma cell lines tested and, to a lower extent, melanocytes, in the context of human histocompatibility leukocyte antigen A2 (HLA-A2), but it did not recognize other tumor cell types. The gene coding for the antigen recognized by this clone was identified by the screening of a melanoma complementary DNA expression library. This antigen is overexpressed in melanomas, compared with other cancer cell lines and healthy tissues, and was thus called melanoma-overexpressed antigen (meloe). Remarkably, the structure of meloe was unusual, with multiple short open reading frames (ORFs). The peptide recognized by the CTL clone was encoded by one of these ORFs, called MELOE-1. Using a specific HLA-A2/peptide tetramer, we showed a correlation between the infusion of TILs containing MELOE-1-specific T cells and relapse prevention in HLA-A2 patients. Indeed, 5 out of 9 patients who did not relapse were infused with TILs that contained MELOE-1-specific T cells, whereas 0 out of the 21 patients who relapsed was infused with such TIL-containing lymphocytes. Overall, our results suggest that this new antigen is involved in immunosurveillance and, thus, represents an attractive target for immunotherapy protocols of melanoma.

Show MeSH
Related in: MedlinePlus