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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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Reactivity of MELOE-1/A2–specific TILs against HLA-A2 tumor cell lines. (A) Lysis of the M170 melanoma cell line (closed circles) and of the 1355 lung carcinoma cell line (open circles) by the M170.48 CTL clone and MELOE-1–specific TIL populations. 51Cr-labeled tumor cells were co-cultured with T cells at various E/T ratios. Chromium release in the supernatants was measured after a 4-h incubation period. (B) Cytokine production by the M170.48 CTL clone and MELOE-1–specific TIL populations in response to M170 melanoma cells. Effector and target cells were incubated at a 1:2 ratio in the presence of Brefeldin A and stained with anti-TNF antibody (open bars), anti– IFN-γ antibody (hatched bars), or anti–IL-2 antibody (closed bars), and 104 T cells were analyzed by flow cytometry.
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fig6: Reactivity of MELOE-1/A2–specific TILs against HLA-A2 tumor cell lines. (A) Lysis of the M170 melanoma cell line (closed circles) and of the 1355 lung carcinoma cell line (open circles) by the M170.48 CTL clone and MELOE-1–specific TIL populations. 51Cr-labeled tumor cells were co-cultured with T cells at various E/T ratios. Chromium release in the supernatants was measured after a 4-h incubation period. (B) Cytokine production by the M170.48 CTL clone and MELOE-1–specific TIL populations in response to M170 melanoma cells. Effector and target cells were incubated at a 1:2 ratio in the presence of Brefeldin A and stained with anti-TNF antibody (open bars), anti– IFN-γ antibody (hatched bars), or anti–IL-2 antibody (closed bars), and 104 T cells were analyzed by flow cytometry.

Mentions: Finally, to address the question of the diversity and tumor reactivity of the MELOE-1/A2–specific repertoire, specific lymphocytes were sorted by monomer-based immunomagnetic sorting (23) from the five positive TIL populations. Fig. 5 B (insets) illustrates the purity of sorted TILs assessed by specific tetramer labeling. We also attempted to sort five negative TIL populations with monomer-coated beads, but no MELOE-1–specific cells were obtained (unpublished data). This last result formally documented the absence of such cells in those populations, or at least showed that the frequencies of MELOE-1–specific T cells were too low to allow their purification by multimer sorting. The diversity of TCR Vβ usage of sorted populations was assessed with a panel of 25 anti-Vβ antibodies representing the most frequently expressed Vβ chains within a normal repertoire. In M117, M170, and M278 sorted populations, eight or six different Vβ chains were significantly expressed (>1%) by MELOE-1/A2–specific TILs, indicating the presence of a rather polyclonal-specific TCR repertoire (Fig. 5 B). TCR diversity of M134 sorted TILs was much lower, with a strong dominance of lymphocytes expressing the Vβ13.1 chain (Fig. 5 B). This may be related to the low fraction of MELOE-1–specific T cells present in this population before sorting (0.3% of CD8+ TILs; Fig. 5 A), which was probably poorly diverse. Finally, we could not determine the dominant Vβ chain expressed by TILs sorted from M180 with our panel of antibodies. Therefore, no dominant Vβ usage could be observed within these three sorted TIL populations. Finally, to support the potential role of MELOE-1/A2–specific TIL transfer in relapse prevention, we studied the reactivity of sorted TIL populations on HLA-A*0201 melanoma cell lines that spontaneously express the MELOE-1 antigen. All sorted T cell lines were lytic against melanoma cell lines (Fig. 6 A and not depicted) and produced IFN-γ and TNF upon stimulation by these cells, with levels similar to the M170.48 CTL clone, and to a lower extent IL-2 (Fig. 6 B and not depicted).


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)

Reactivity of MELOE-1/A2–specific TILs against HLA-A2 tumor cell lines. (A) Lysis of the M170 melanoma cell line (closed circles) and of the 1355 lung carcinoma cell line (open circles) by the M170.48 CTL clone and MELOE-1–specific TIL populations. 51Cr-labeled tumor cells were co-cultured with T cells at various E/T ratios. Chromium release in the supernatants was measured after a 4-h incubation period. (B) Cytokine production by the M170.48 CTL clone and MELOE-1–specific TIL populations in response to M170 melanoma cells. Effector and target cells were incubated at a 1:2 ratio in the presence of Brefeldin A and stained with anti-TNF antibody (open bars), anti– IFN-γ antibody (hatched bars), or anti–IL-2 antibody (closed bars), and 104 T cells were analyzed by flow cytometry.
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fig6: Reactivity of MELOE-1/A2–specific TILs against HLA-A2 tumor cell lines. (A) Lysis of the M170 melanoma cell line (closed circles) and of the 1355 lung carcinoma cell line (open circles) by the M170.48 CTL clone and MELOE-1–specific TIL populations. 51Cr-labeled tumor cells were co-cultured with T cells at various E/T ratios. Chromium release in the supernatants was measured after a 4-h incubation period. (B) Cytokine production by the M170.48 CTL clone and MELOE-1–specific TIL populations in response to M170 melanoma cells. Effector and target cells were incubated at a 1:2 ratio in the presence of Brefeldin A and stained with anti-TNF antibody (open bars), anti– IFN-γ antibody (hatched bars), or anti–IL-2 antibody (closed bars), and 104 T cells were analyzed by flow cytometry.
Mentions: Finally, to address the question of the diversity and tumor reactivity of the MELOE-1/A2–specific repertoire, specific lymphocytes were sorted by monomer-based immunomagnetic sorting (23) from the five positive TIL populations. Fig. 5 B (insets) illustrates the purity of sorted TILs assessed by specific tetramer labeling. We also attempted to sort five negative TIL populations with monomer-coated beads, but no MELOE-1–specific cells were obtained (unpublished data). This last result formally documented the absence of such cells in those populations, or at least showed that the frequencies of MELOE-1–specific T cells were too low to allow their purification by multimer sorting. The diversity of TCR Vβ usage of sorted populations was assessed with a panel of 25 anti-Vβ antibodies representing the most frequently expressed Vβ chains within a normal repertoire. In M117, M170, and M278 sorted populations, eight or six different Vβ chains were significantly expressed (>1%) by MELOE-1/A2–specific TILs, indicating the presence of a rather polyclonal-specific TCR repertoire (Fig. 5 B). TCR diversity of M134 sorted TILs was much lower, with a strong dominance of lymphocytes expressing the Vβ13.1 chain (Fig. 5 B). This may be related to the low fraction of MELOE-1–specific T cells present in this population before sorting (0.3% of CD8+ TILs; Fig. 5 A), which was probably poorly diverse. Finally, we could not determine the dominant Vβ chain expressed by TILs sorted from M180 with our panel of antibodies. Therefore, no dominant Vβ usage could be observed within these three sorted TIL populations. Finally, to support the potential role of MELOE-1/A2–specific TIL transfer in relapse prevention, we studied the reactivity of sorted TIL populations on HLA-A*0201 melanoma cell lines that spontaneously express the MELOE-1 antigen. All sorted T cell lines were lytic against melanoma cell lines (Fig. 6 A and not depicted) and produced IFN-γ and TNF upon stimulation by these cells, with levels similar to the M170.48 CTL clone, and to a lower extent IL-2 (Fig. 6 B and not depicted).

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