T Cell Receptor (TCR) structure of autologous melanoma-reactive Cytotoxic T Lymphocyte (CTL) clones: Tumor-infiltrating lymphocytes overexpress in vivo the TCR β chain sequence used by an HLA-A2-restricted and melanocyte-lineage-specific CTL clone

Marialuisa Sensi, Stefania Salvi, Chiara Castelli, Cristina Maccalli, Arabella Mazzocchi, Roberta Mortarini, Gabriella Nicolini, Meenhard Herlyn, Giorgio Parmiani, Andrea Anichini

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Abstract

HLA-A2+ melanomas express common melanoma-associated antigens (Ags) recognized in vitro by autologous cytotoxic T lymphocytes (CTL). However, it is not known whether tumor Ags can drive in vivo a selective accumulation/expansion of Ag-specific, tumor-infiltrating T lymphocytes (TIL). Therefore, to evaluate this possibility, 39 CTL clones isolated from several independent mixed lymphocyte tumor cultures (MLTC) of TIL and peripheral blood lymphocytes (PBL) of an HLA-A2+ melanoma patient and selected for T cell receptor (TCR)-dependent, HLA-restricted tumor lysis, were used for analysis of TCR α and β chain structure by the cDNA polymerase chain reaction (PCR) technique with variable gene-specific primers followed by sequencing. Despite absence of oligoclonality in fresh TIL and PBL, as well as in T cells of day 28 MLTC (day of cloning), sequence analysis of TCR α and β chains of TIL clones revealed a dominance of a major category of melanoma-specific, HLA-A2-restricted T cells expressing a Vα8.2/JαAP511/Cα and Vβ2.1/Dβ1/Jβ1.1/Cβ1 TCR. The same TCR was also found in 2 out of 14 PBL clones. The other PBL clones employed a Vα2.1 gene segment associated with either Vβ13.2, 14, or w22. Clones A81 (Vα2.1/JαIGRJα04/Cα and Vβ14/Dβ1/Jβ1.2/Cβ1) and A21 (Vα8.2/JαAP511/Cα and Vβ2.1/Dβ1/Jβ1.1/Cβ1), representative of the two most frequent TCR of PBL and TIL, respectively, expressed different lytic patterns, but both were HLA-A2 restricted and lysed only HLA-A2+ melanomas and normal melanocytes, thus indicating recognition of two distinct HLA-A2-associated and tissue-related Ags. Finally, by the inverse PCR technique, the specific TCR β chain (Vβ2.1/Dβ1/Jβ1.1/Cβ1) expressed by the dominant TIL clone was found to represent 19 and 18.4% of all Vβ2 sequences expressed in the fresh tumor sample and in the purified TIL, respectively, but <0.19% of Vβ2+ sequences expressed in PBL. These results are consistent with the hypothesis that a clonal expansion/accumulation of a melanocyte-lineage-specific and HLA-A2-restricted T cell clone occurred in vivo at the site of tumor growth.

Original languageEnglish
Pages (from-to)1231-1246
Number of pages16
JournalJournal of Experimental Medicine
Volume178
Issue number4
Publication statusPublished - 1 Oct 1993
Externally publishedYes

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Tumor-Infiltrating Lymphocytes
HLA-A2 Antigen
Melanocytes
Cytotoxic T-Lymphocytes
T-Cell Antigen Receptor
Melanoma
Clone Cells
T-Lymphocytes
Lymphocytes
Neoplasms
Melanoma-Specific Antigens
Polymerase Chain Reaction
Neoplasm Antigens
Genes
Sequence Analysis
Organism Cloning
Complementary DNA

ASJC Scopus subject areas

  • Immunology

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T Cell Receptor (TCR) structure of autologous melanoma-reactive Cytotoxic T Lymphocyte (CTL) clones : Tumor-infiltrating lymphocytes overexpress in vivo the TCR β chain sequence used by an HLA-A2-restricted and melanocyte-lineage-specific CTL clone. / Sensi, Marialuisa; Salvi, Stefania; Castelli, Chiara; Maccalli, Cristina; Mazzocchi, Arabella; Mortarini, Roberta; Nicolini, Gabriella; Herlyn, Meenhard; Parmiani, Giorgio; Anichini, Andrea.

In: Journal of Experimental Medicine, Vol. 178, No. 4, 01.10.1993, p. 1231-1246.

Research output: Contribution to journalArticle

Sensi, Marialuisa ; Salvi, Stefania ; Castelli, Chiara ; Maccalli, Cristina ; Mazzocchi, Arabella ; Mortarini, Roberta ; Nicolini, Gabriella ; Herlyn, Meenhard ; Parmiani, Giorgio ; Anichini, Andrea. / T Cell Receptor (TCR) structure of autologous melanoma-reactive Cytotoxic T Lymphocyte (CTL) clones : Tumor-infiltrating lymphocytes overexpress in vivo the TCR β chain sequence used by an HLA-A2-restricted and melanocyte-lineage-specific CTL clone. In: Journal of Experimental Medicine. 1993 ; Vol. 178, No. 4. pp. 1231-1246.
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abstract = "HLA-A2+ melanomas express common melanoma-associated antigens (Ags) recognized in vitro by autologous cytotoxic T lymphocytes (CTL). However, it is not known whether tumor Ags can drive in vivo a selective accumulation/expansion of Ag-specific, tumor-infiltrating T lymphocytes (TIL). Therefore, to evaluate this possibility, 39 CTL clones isolated from several independent mixed lymphocyte tumor cultures (MLTC) of TIL and peripheral blood lymphocytes (PBL) of an HLA-A2+ melanoma patient and selected for T cell receptor (TCR)-dependent, HLA-restricted tumor lysis, were used for analysis of TCR α and β chain structure by the cDNA polymerase chain reaction (PCR) technique with variable gene-specific primers followed by sequencing. Despite absence of oligoclonality in fresh TIL and PBL, as well as in T cells of day 28 MLTC (day of cloning), sequence analysis of TCR α and β chains of TIL clones revealed a dominance of a major category of melanoma-specific, HLA-A2-restricted T cells expressing a Vα8.2/JαAP511/Cα and Vβ2.1/Dβ1/Jβ1.1/Cβ1 TCR. The same TCR was also found in 2 out of 14 PBL clones. The other PBL clones employed a Vα2.1 gene segment associated with either Vβ13.2, 14, or w22. Clones A81 (Vα2.1/JαIGRJα04/Cα and Vβ14/Dβ1/Jβ1.2/Cβ1) and A21 (Vα8.2/JαAP511/Cα and Vβ2.1/Dβ1/Jβ1.1/Cβ1), representative of the two most frequent TCR of PBL and TIL, respectively, expressed different lytic patterns, but both were HLA-A2 restricted and lysed only HLA-A2+ melanomas and normal melanocytes, thus indicating recognition of two distinct HLA-A2-associated and tissue-related Ags. Finally, by the inverse PCR technique, the specific TCR β chain (Vβ2.1/Dβ1/Jβ1.1/Cβ1) expressed by the dominant TIL clone was found to represent 19 and 18.4{\%} of all Vβ2 sequences expressed in the fresh tumor sample and in the purified TIL, respectively, but <0.19{\%} of Vβ2+ sequences expressed in PBL. These results are consistent with the hypothesis that a clonal expansion/accumulation of a melanocyte-lineage-specific and HLA-A2-restricted T cell clone occurred in vivo at the site of tumor growth.",
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T1 - T Cell Receptor (TCR) structure of autologous melanoma-reactive Cytotoxic T Lymphocyte (CTL) clones

T2 - Tumor-infiltrating lymphocytes overexpress in vivo the TCR β chain sequence used by an HLA-A2-restricted and melanocyte-lineage-specific CTL clone

AU - Sensi, Marialuisa

AU - Salvi, Stefania

AU - Castelli, Chiara

AU - Maccalli, Cristina

AU - Mazzocchi, Arabella

AU - Mortarini, Roberta

AU - Nicolini, Gabriella

AU - Herlyn, Meenhard

AU - Parmiani, Giorgio

AU - Anichini, Andrea

PY - 1993/10/1

Y1 - 1993/10/1

N2 - HLA-A2+ melanomas express common melanoma-associated antigens (Ags) recognized in vitro by autologous cytotoxic T lymphocytes (CTL). However, it is not known whether tumor Ags can drive in vivo a selective accumulation/expansion of Ag-specific, tumor-infiltrating T lymphocytes (TIL). Therefore, to evaluate this possibility, 39 CTL clones isolated from several independent mixed lymphocyte tumor cultures (MLTC) of TIL and peripheral blood lymphocytes (PBL) of an HLA-A2+ melanoma patient and selected for T cell receptor (TCR)-dependent, HLA-restricted tumor lysis, were used for analysis of TCR α and β chain structure by the cDNA polymerase chain reaction (PCR) technique with variable gene-specific primers followed by sequencing. Despite absence of oligoclonality in fresh TIL and PBL, as well as in T cells of day 28 MLTC (day of cloning), sequence analysis of TCR α and β chains of TIL clones revealed a dominance of a major category of melanoma-specific, HLA-A2-restricted T cells expressing a Vα8.2/JαAP511/Cα and Vβ2.1/Dβ1/Jβ1.1/Cβ1 TCR. The same TCR was also found in 2 out of 14 PBL clones. The other PBL clones employed a Vα2.1 gene segment associated with either Vβ13.2, 14, or w22. Clones A81 (Vα2.1/JαIGRJα04/Cα and Vβ14/Dβ1/Jβ1.2/Cβ1) and A21 (Vα8.2/JαAP511/Cα and Vβ2.1/Dβ1/Jβ1.1/Cβ1), representative of the two most frequent TCR of PBL and TIL, respectively, expressed different lytic patterns, but both were HLA-A2 restricted and lysed only HLA-A2+ melanomas and normal melanocytes, thus indicating recognition of two distinct HLA-A2-associated and tissue-related Ags. Finally, by the inverse PCR technique, the specific TCR β chain (Vβ2.1/Dβ1/Jβ1.1/Cβ1) expressed by the dominant TIL clone was found to represent 19 and 18.4% of all Vβ2 sequences expressed in the fresh tumor sample and in the purified TIL, respectively, but <0.19% of Vβ2+ sequences expressed in PBL. These results are consistent with the hypothesis that a clonal expansion/accumulation of a melanocyte-lineage-specific and HLA-A2-restricted T cell clone occurred in vivo at the site of tumor growth.

AB - HLA-A2+ melanomas express common melanoma-associated antigens (Ags) recognized in vitro by autologous cytotoxic T lymphocytes (CTL). However, it is not known whether tumor Ags can drive in vivo a selective accumulation/expansion of Ag-specific, tumor-infiltrating T lymphocytes (TIL). Therefore, to evaluate this possibility, 39 CTL clones isolated from several independent mixed lymphocyte tumor cultures (MLTC) of TIL and peripheral blood lymphocytes (PBL) of an HLA-A2+ melanoma patient and selected for T cell receptor (TCR)-dependent, HLA-restricted tumor lysis, were used for analysis of TCR α and β chain structure by the cDNA polymerase chain reaction (PCR) technique with variable gene-specific primers followed by sequencing. Despite absence of oligoclonality in fresh TIL and PBL, as well as in T cells of day 28 MLTC (day of cloning), sequence analysis of TCR α and β chains of TIL clones revealed a dominance of a major category of melanoma-specific, HLA-A2-restricted T cells expressing a Vα8.2/JαAP511/Cα and Vβ2.1/Dβ1/Jβ1.1/Cβ1 TCR. The same TCR was also found in 2 out of 14 PBL clones. The other PBL clones employed a Vα2.1 gene segment associated with either Vβ13.2, 14, or w22. Clones A81 (Vα2.1/JαIGRJα04/Cα and Vβ14/Dβ1/Jβ1.2/Cβ1) and A21 (Vα8.2/JαAP511/Cα and Vβ2.1/Dβ1/Jβ1.1/Cβ1), representative of the two most frequent TCR of PBL and TIL, respectively, expressed different lytic patterns, but both were HLA-A2 restricted and lysed only HLA-A2+ melanomas and normal melanocytes, thus indicating recognition of two distinct HLA-A2-associated and tissue-related Ags. Finally, by the inverse PCR technique, the specific TCR β chain (Vβ2.1/Dβ1/Jβ1.1/Cβ1) expressed by the dominant TIL clone was found to represent 19 and 18.4% of all Vβ2 sequences expressed in the fresh tumor sample and in the purified TIL, respectively, but <0.19% of Vβ2+ sequences expressed in PBL. These results are consistent with the hypothesis that a clonal expansion/accumulation of a melanocyte-lineage-specific and HLA-A2-restricted T cell clone occurred in vivo at the site of tumor growth.

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