Functional and molecular characterization of tumour-infiltrating lymphocytes and clones thereof from a major-histocompatibility-complex-negative human tumour

Neuroblastoma

Paola Facchetti, Ignazia Prigione, Fabio Ghiotto, Paola Tasso, Alberto Garaventa, Vito Pistoia

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Neuroblastoma (NB) is a major-histocompatibility-complex (MHC)-negative neuroectodermal tumour that is often infiltrated with lymphocytes. A detailed characterization of NB-associated tumour-infiltrating lymphocytes (TIL) has never been carried out. Here we have investigated the immmunophenotype and the cytotoxic activities of TIL from nine and seven NB patients respectively. Furthermore, the T cell receptor (TcR) variability and the patterns of cytokine gene expression of fresh versus recombinant (r) interleukin(IL)-2-cultured TIL were studied in four NB cases. The results obtained showed the following: (1) freshly isolated TIL were comprised of a mixture of CD4+ and CD8+ T cells partially expressing HLA-DR and/or CD25. The CD4/CD8 ratio ranged from 0.5 to 5 in the different cases. Upon culture of TIL with rIL-2, an increased proportion of CD56+ and CD8+ lymphocytes was consistently observed; (2) IL-2-expanded TIL lysed natural killer (NK)-sensitive and lymphokine-activated-killer(LAK)-sensitive target cell lines; (3) reverse-transcriptase/polymerase-chain-reaction (RT-PCR) experiments showed that most TcR Vβ genes were expressed both in fresh and in cultured TIL, suggesting that such cell populations were polyclonal; (4) interferon γ, IL-4, IL-5, tumour necrosis factor (TNF) α, IL-8, IL-10 mRNA and, to a lesser extent, IL-2 mRNA were expressed by cultured TIL, as assessed by RT-PCR; the corresponding tumour samples consistently contained TNFα, IL-8 and IL-10 mRNA, whereas IL-2 and IFNγ mRNA were faintly expressed in some NB tumors and IL-4 and IL-5 mRNA were never detected. A total of 90 clones were subsequently raised from IL-2-expanded TIL from six NB patients; 87/90 clones were of T cell lineage with a CD4+ or CD8+ immunophenotype, whereas the 3 remaining clones were of NK cell origin. Upon triggering of the CD3-TcR complex, 64% CD4+ and 77% CD8+ T cell clones killed the murine P815 mastocytoma cell line. Virtually no T cell clone lysed a LAK-sensitive NB cell line, whereas 15% CD4+ and 17% CD8+ clones mediated NK-like activity against the K562 cell line. Finally, the patterns of cytokine production by CD4+ clones were roughly consistent with those of a T helper (Th)1 profile and similar to those observed in CD8+ clones.

Original languageEnglish
Pages (from-to)170-178
Number of pages9
JournalCancer Immunology, Immunotherapy
Volume42
Issue number3
DOIs
Publication statusPublished - 1996

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Tumor-Infiltrating Lymphocytes
Major Histocompatibility Complex
Neuroblastoma
Clone Cells
Interleukin-2
Neoplasms
Messenger RNA
T-Lymphocytes
Cell Line
Lymphokines
Interleukin-5
T-Cell Antigen Receptor
Reverse Transcriptase Polymerase Chain Reaction
Interleukin-8
Interleukin-4
Interleukin-10
Tumor Necrosis Factor-alpha
Neuroectodermal Tumors
Lymphocytes
Mastocytoma

Keywords

  • cytokines
  • cytotoxicity
  • neuroblastoma
  • T cell receptor
  • TIL

ASJC Scopus subject areas

  • Cancer Research
  • Immunology
  • Oncology

Cite this

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title = "Functional and molecular characterization of tumour-infiltrating lymphocytes and clones thereof from a major-histocompatibility-complex-negative human tumour: Neuroblastoma",
abstract = "Neuroblastoma (NB) is a major-histocompatibility-complex (MHC)-negative neuroectodermal tumour that is often infiltrated with lymphocytes. A detailed characterization of NB-associated tumour-infiltrating lymphocytes (TIL) has never been carried out. Here we have investigated the immmunophenotype and the cytotoxic activities of TIL from nine and seven NB patients respectively. Furthermore, the T cell receptor (TcR) variability and the patterns of cytokine gene expression of fresh versus recombinant (r) interleukin(IL)-2-cultured TIL were studied in four NB cases. The results obtained showed the following: (1) freshly isolated TIL were comprised of a mixture of CD4+ and CD8+ T cells partially expressing HLA-DR and/or CD25. The CD4/CD8 ratio ranged from 0.5 to 5 in the different cases. Upon culture of TIL with rIL-2, an increased proportion of CD56+ and CD8+ lymphocytes was consistently observed; (2) IL-2-expanded TIL lysed natural killer (NK)-sensitive and lymphokine-activated-killer(LAK)-sensitive target cell lines; (3) reverse-transcriptase/polymerase-chain-reaction (RT-PCR) experiments showed that most TcR Vβ genes were expressed both in fresh and in cultured TIL, suggesting that such cell populations were polyclonal; (4) interferon γ, IL-4, IL-5, tumour necrosis factor (TNF) α, IL-8, IL-10 mRNA and, to a lesser extent, IL-2 mRNA were expressed by cultured TIL, as assessed by RT-PCR; the corresponding tumour samples consistently contained TNFα, IL-8 and IL-10 mRNA, whereas IL-2 and IFNγ mRNA were faintly expressed in some NB tumors and IL-4 and IL-5 mRNA were never detected. A total of 90 clones were subsequently raised from IL-2-expanded TIL from six NB patients; 87/90 clones were of T cell lineage with a CD4+ or CD8+ immunophenotype, whereas the 3 remaining clones were of NK cell origin. Upon triggering of the CD3-TcR complex, 64{\%} CD4+ and 77{\%} CD8+ T cell clones killed the murine P815 mastocytoma cell line. Virtually no T cell clone lysed a LAK-sensitive NB cell line, whereas 15{\%} CD4+ and 17{\%} CD8+ clones mediated NK-like activity against the K562 cell line. Finally, the patterns of cytokine production by CD4+ clones were roughly consistent with those of a T helper (Th)1 profile and similar to those observed in CD8+ clones.",
keywords = "cytokines, cytotoxicity, neuroblastoma, T cell receptor, TIL",
author = "Paola Facchetti and Ignazia Prigione and Fabio Ghiotto and Paola Tasso and Alberto Garaventa and Vito Pistoia",
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T1 - Functional and molecular characterization of tumour-infiltrating lymphocytes and clones thereof from a major-histocompatibility-complex-negative human tumour

T2 - Neuroblastoma

AU - Facchetti, Paola

AU - Prigione, Ignazia

AU - Ghiotto, Fabio

AU - Tasso, Paola

AU - Garaventa, Alberto

AU - Pistoia, Vito

PY - 1996

Y1 - 1996

N2 - Neuroblastoma (NB) is a major-histocompatibility-complex (MHC)-negative neuroectodermal tumour that is often infiltrated with lymphocytes. A detailed characterization of NB-associated tumour-infiltrating lymphocytes (TIL) has never been carried out. Here we have investigated the immmunophenotype and the cytotoxic activities of TIL from nine and seven NB patients respectively. Furthermore, the T cell receptor (TcR) variability and the patterns of cytokine gene expression of fresh versus recombinant (r) interleukin(IL)-2-cultured TIL were studied in four NB cases. The results obtained showed the following: (1) freshly isolated TIL were comprised of a mixture of CD4+ and CD8+ T cells partially expressing HLA-DR and/or CD25. The CD4/CD8 ratio ranged from 0.5 to 5 in the different cases. Upon culture of TIL with rIL-2, an increased proportion of CD56+ and CD8+ lymphocytes was consistently observed; (2) IL-2-expanded TIL lysed natural killer (NK)-sensitive and lymphokine-activated-killer(LAK)-sensitive target cell lines; (3) reverse-transcriptase/polymerase-chain-reaction (RT-PCR) experiments showed that most TcR Vβ genes were expressed both in fresh and in cultured TIL, suggesting that such cell populations were polyclonal; (4) interferon γ, IL-4, IL-5, tumour necrosis factor (TNF) α, IL-8, IL-10 mRNA and, to a lesser extent, IL-2 mRNA were expressed by cultured TIL, as assessed by RT-PCR; the corresponding tumour samples consistently contained TNFα, IL-8 and IL-10 mRNA, whereas IL-2 and IFNγ mRNA were faintly expressed in some NB tumors and IL-4 and IL-5 mRNA were never detected. A total of 90 clones were subsequently raised from IL-2-expanded TIL from six NB patients; 87/90 clones were of T cell lineage with a CD4+ or CD8+ immunophenotype, whereas the 3 remaining clones were of NK cell origin. Upon triggering of the CD3-TcR complex, 64% CD4+ and 77% CD8+ T cell clones killed the murine P815 mastocytoma cell line. Virtually no T cell clone lysed a LAK-sensitive NB cell line, whereas 15% CD4+ and 17% CD8+ clones mediated NK-like activity against the K562 cell line. Finally, the patterns of cytokine production by CD4+ clones were roughly consistent with those of a T helper (Th)1 profile and similar to those observed in CD8+ clones.

AB - Neuroblastoma (NB) is a major-histocompatibility-complex (MHC)-negative neuroectodermal tumour that is often infiltrated with lymphocytes. A detailed characterization of NB-associated tumour-infiltrating lymphocytes (TIL) has never been carried out. Here we have investigated the immmunophenotype and the cytotoxic activities of TIL from nine and seven NB patients respectively. Furthermore, the T cell receptor (TcR) variability and the patterns of cytokine gene expression of fresh versus recombinant (r) interleukin(IL)-2-cultured TIL were studied in four NB cases. The results obtained showed the following: (1) freshly isolated TIL were comprised of a mixture of CD4+ and CD8+ T cells partially expressing HLA-DR and/or CD25. The CD4/CD8 ratio ranged from 0.5 to 5 in the different cases. Upon culture of TIL with rIL-2, an increased proportion of CD56+ and CD8+ lymphocytes was consistently observed; (2) IL-2-expanded TIL lysed natural killer (NK)-sensitive and lymphokine-activated-killer(LAK)-sensitive target cell lines; (3) reverse-transcriptase/polymerase-chain-reaction (RT-PCR) experiments showed that most TcR Vβ genes were expressed both in fresh and in cultured TIL, suggesting that such cell populations were polyclonal; (4) interferon γ, IL-4, IL-5, tumour necrosis factor (TNF) α, IL-8, IL-10 mRNA and, to a lesser extent, IL-2 mRNA were expressed by cultured TIL, as assessed by RT-PCR; the corresponding tumour samples consistently contained TNFα, IL-8 and IL-10 mRNA, whereas IL-2 and IFNγ mRNA were faintly expressed in some NB tumors and IL-4 and IL-5 mRNA were never detected. A total of 90 clones were subsequently raised from IL-2-expanded TIL from six NB patients; 87/90 clones were of T cell lineage with a CD4+ or CD8+ immunophenotype, whereas the 3 remaining clones were of NK cell origin. Upon triggering of the CD3-TcR complex, 64% CD4+ and 77% CD8+ T cell clones killed the murine P815 mastocytoma cell line. Virtually no T cell clone lysed a LAK-sensitive NB cell line, whereas 15% CD4+ and 17% CD8+ clones mediated NK-like activity against the K562 cell line. Finally, the patterns of cytokine production by CD4+ clones were roughly consistent with those of a T helper (Th)1 profile and similar to those observed in CD8+ clones.

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KW - neuroblastoma

KW - T cell receptor

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