Functional CD4(+) and CD8(+) T-cell responses induced by autologous mitomycin C treated Epstein-Barr virus transformed lymphoblastoid cell lines

P Meij, E Bloemena, N Palmen, A Brink, M B Vervoort, C J Meijer, J M Middeldorp

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Epstein-Barr virus (EBV) gene expression in tumor cells of posttransplant lymphoproliferative disorder (PTLD) patients resembles that of EBV transformed B-cell lines (LCL). EBV-specific cytotoxic T-lymphocytes can be generated by stimulating peripheral blood lymphocytes with autologous LCL. We describe a standardized method for the growth inactivation and cryopreservation of LCL for optimal T-cell stimulation and analyzed the function and phenotype of responding T-cells. LCL growth was completely blocked by mitomycin C treatment (McLCL) and McLCL could be cryopreserved while retaining excellent APC function. McLCL stimulated both CD4(+) and CD8(+) T-cells as measured by HLA-DR and CD25 expression using FACS analysis. EBV-specific CTL activity and T-cell proliferation were induced and immunocytochemical staining showed CD4(+) and (granzyme B positive) CD8(+) T-cells rosetting with McLCL. Granzymes A and B, IFN-gamma, and IL-6 were detected at significant levels in the supernatant. Thus, ex vivo T-cell activation with cryopreserved McLCL results in activation of both CD4(+) and CD8(+) T-cells producing a Th1-like cytokine profile, making this a suitable protocol for adoptive therapy of PTLD.

Original languageEnglish
Pages (from-to)25-33
Number of pages9
JournalCellular Immunology
Volume208
Issue number1
DOIs
Publication statusPublished - 25 Feb 2001

Cite this

@article{f9fe5e17fc7a43e1b20b1fcf31bad1a7,
title = "Functional CD4(+) and CD8(+) T-cell responses induced by autologous mitomycin C treated Epstein-Barr virus transformed lymphoblastoid cell lines",
abstract = "Epstein-Barr virus (EBV) gene expression in tumor cells of posttransplant lymphoproliferative disorder (PTLD) patients resembles that of EBV transformed B-cell lines (LCL). EBV-specific cytotoxic T-lymphocytes can be generated by stimulating peripheral blood lymphocytes with autologous LCL. We describe a standardized method for the growth inactivation and cryopreservation of LCL for optimal T-cell stimulation and analyzed the function and phenotype of responding T-cells. LCL growth was completely blocked by mitomycin C treatment (McLCL) and McLCL could be cryopreserved while retaining excellent APC function. McLCL stimulated both CD4(+) and CD8(+) T-cells as measured by HLA-DR and CD25 expression using FACS analysis. EBV-specific CTL activity and T-cell proliferation were induced and immunocytochemical staining showed CD4(+) and (granzyme B positive) CD8(+) T-cells rosetting with McLCL. Granzymes A and B, IFN-gamma, and IL-6 were detected at significant levels in the supernatant. Thus, ex vivo T-cell activation with cryopreserved McLCL results in activation of both CD4(+) and CD8(+) T-cells producing a Th1-like cytokine profile, making this a suitable protocol for adoptive therapy of PTLD.",
keywords = "Adoptive Transfer, Antigen Presentation, CD4-Positive T-Lymphocytes, CD8-Positive T-Lymphocytes, Cell Division, Cell Line, Transformed, Cryopreservation, Cytotoxicity, Immunologic, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Granzymes, Herpesvirus 4, Human, Humans, Interferon-gamma, Interleukin-6, Lymphocyte Activation, Mitomycin, Phenotype, Rosette Formation, Serine Endopeptidases, T-Lymphocytes, Cytotoxic, Tumor Cells, Cultured, Journal Article",
author = "P Meij and E Bloemena and N Palmen and A Brink and Vervoort, {M B} and Meijer, {C J} and Middeldorp, {J M}",
note = "Copyright 2001 Academic Press.",
year = "2001",
month = "2",
day = "25",
doi = "10.1006/cimm.2001.1760",
language = "English",
volume = "208",
pages = "25--33",
journal = "Cellular Immunology",
issn = "0008-8749",
publisher = "Academic Press Inc.",
number = "1",

}

Functional CD4(+) and CD8(+) T-cell responses induced by autologous mitomycin C treated Epstein-Barr virus transformed lymphoblastoid cell lines. / Meij, P; Bloemena, E; Palmen, N; Brink, A; Vervoort, M B; Meijer, C J; Middeldorp, J M.

In: Cellular Immunology, Vol. 208, No. 1, 25.02.2001, p. 25-33.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Functional CD4(+) and CD8(+) T-cell responses induced by autologous mitomycin C treated Epstein-Barr virus transformed lymphoblastoid cell lines

AU - Meij, P

AU - Bloemena, E

AU - Palmen, N

AU - Brink, A

AU - Vervoort, M B

AU - Meijer, C J

AU - Middeldorp, J M

N1 - Copyright 2001 Academic Press.

PY - 2001/2/25

Y1 - 2001/2/25

N2 - Epstein-Barr virus (EBV) gene expression in tumor cells of posttransplant lymphoproliferative disorder (PTLD) patients resembles that of EBV transformed B-cell lines (LCL). EBV-specific cytotoxic T-lymphocytes can be generated by stimulating peripheral blood lymphocytes with autologous LCL. We describe a standardized method for the growth inactivation and cryopreservation of LCL for optimal T-cell stimulation and analyzed the function and phenotype of responding T-cells. LCL growth was completely blocked by mitomycin C treatment (McLCL) and McLCL could be cryopreserved while retaining excellent APC function. McLCL stimulated both CD4(+) and CD8(+) T-cells as measured by HLA-DR and CD25 expression using FACS analysis. EBV-specific CTL activity and T-cell proliferation were induced and immunocytochemical staining showed CD4(+) and (granzyme B positive) CD8(+) T-cells rosetting with McLCL. Granzymes A and B, IFN-gamma, and IL-6 were detected at significant levels in the supernatant. Thus, ex vivo T-cell activation with cryopreserved McLCL results in activation of both CD4(+) and CD8(+) T-cells producing a Th1-like cytokine profile, making this a suitable protocol for adoptive therapy of PTLD.

AB - Epstein-Barr virus (EBV) gene expression in tumor cells of posttransplant lymphoproliferative disorder (PTLD) patients resembles that of EBV transformed B-cell lines (LCL). EBV-specific cytotoxic T-lymphocytes can be generated by stimulating peripheral blood lymphocytes with autologous LCL. We describe a standardized method for the growth inactivation and cryopreservation of LCL for optimal T-cell stimulation and analyzed the function and phenotype of responding T-cells. LCL growth was completely blocked by mitomycin C treatment (McLCL) and McLCL could be cryopreserved while retaining excellent APC function. McLCL stimulated both CD4(+) and CD8(+) T-cells as measured by HLA-DR and CD25 expression using FACS analysis. EBV-specific CTL activity and T-cell proliferation were induced and immunocytochemical staining showed CD4(+) and (granzyme B positive) CD8(+) T-cells rosetting with McLCL. Granzymes A and B, IFN-gamma, and IL-6 were detected at significant levels in the supernatant. Thus, ex vivo T-cell activation with cryopreserved McLCL results in activation of both CD4(+) and CD8(+) T-cells producing a Th1-like cytokine profile, making this a suitable protocol for adoptive therapy of PTLD.

KW - Adoptive Transfer

KW - Antigen Presentation

KW - CD4-Positive T-Lymphocytes

KW - CD8-Positive T-Lymphocytes

KW - Cell Division

KW - Cell Line, Transformed

KW - Cryopreservation

KW - Cytotoxicity, Immunologic

KW - Enzyme-Linked Immunosorbent Assay

KW - Flow Cytometry

KW - Granzymes

KW - Herpesvirus 4, Human

KW - Humans

KW - Interferon-gamma

KW - Interleukin-6

KW - Lymphocyte Activation

KW - Mitomycin

KW - Phenotype

KW - Rosette Formation

KW - Serine Endopeptidases

KW - T-Lymphocytes, Cytotoxic

KW - Tumor Cells, Cultured

KW - Journal Article

U2 - 10.1006/cimm.2001.1760

DO - 10.1006/cimm.2001.1760

M3 - Article

VL - 208

SP - 25

EP - 33

JO - Cellular Immunology

JF - Cellular Immunology

SN - 0008-8749

IS - 1

ER -