CD40 ligand-stimulated B cell precursor leukemic cells elicit interferon-γ production by autologous bone marrow T cells in childhood acute lymphoblastic leukemia

E. Todisco, G. Gaipa, E. Biagi, M. Bonamino, R. Gramigna, M. Introna, A. Biondi

Research output: Contribution to journalArticle

Abstract

Childhood B cell precursor acute lymphoblastic leukemia (BCP-ALL) cells, collected from bone marrow (BM) at diagnosis, were cultured, after thawing, on allogeneic human bone marrow stroma (HBMS) for 48 h in the presence of a soluble trimeric CD40 ligand (stCD40L) molecule. HBMS maintained leukemic cells viability in all tested cases (mean viability 85%). Under these culture conditions we noticed upregulation or de novo expression of costimulatory molecules CD40, CD80 (B7-1) and CD86 (B7-2) in 22/22, 15/23 and 21/23 cases, respectively. Upregulation, in terms of fluorescence intensity, was also observed in the expression of MHC I, MHC II, CD54 (ICAM 1) and CD58 (LFA 3) molecules. HBMS alone, although to a lesser extent, was able to induce modulation of these molecules, but not CD80, in a similar proportion of cases. Neither stCD40L nor HBMS induced modulation of CD10 and CD34 molecules. Moreover, in 4/4 tested cases, stCD40L-stimulated ALL cells were able to induce allogeneic T cells proliferation. To evaluate whether leukemia-reactive T cells were detectable in the BM of ALL patients at diagnosis, stCD40L-stimulated ALL cells were co-cultured with autologous T cells (ratio 1:1), isolated from BM at diagnosis, for 4 days and a 24 h ELISPOT assay was applied to detect the presence of interferon-gamma (IFN-γ)-producing cells. In four of seven cases IFN-γ-producing cells were detected with frequencies of 1/900, 1/1560, 1/2150 and 1/1575 autologous T cells. These data confirm that stCD40L exposure can activate the antigen-presenting cell (APC) capacity of BCP-ALL cells cultured on HBMS and that ELISPOT assay can be used to measure the frequency of leukemia-reactive autologous T cells in the BM of ALL patients even after short-term culture with stCD40L-stimulated ALL cells.

Original languageEnglish
Pages (from-to)2046-2054
Number of pages9
JournalLeukemia
Volume16
Issue number10
DOIs
Publication statusPublished - Oct 1 2002

Fingerprint

CD40 Ligand
B-Lymphoid Precursor Cells
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Bone Marrow Cells
Interferons
Bone Marrow
T-Lymphocytes
Enzyme-Linked Immunospot Assay
Interferon-gamma
Up-Regulation
CD58 Antigens
T-Cell Leukemia
Antigen-Presenting Cells
Intercellular Adhesion Molecule-1
Cultured Cells
Cell Survival
Leukemia
Fluorescence
Cell Proliferation

Keywords

  • CD40 ligand
  • Childhood ALL
  • ELISPOT
  • Immunology
  • Interferongamma
  • Stroma

ASJC Scopus subject areas

  • Hematology
  • Cancer Research

Cite this

CD40 ligand-stimulated B cell precursor leukemic cells elicit interferon-γ production by autologous bone marrow T cells in childhood acute lymphoblastic leukemia. / Todisco, E.; Gaipa, G.; Biagi, E.; Bonamino, M.; Gramigna, R.; Introna, M.; Biondi, A.

In: Leukemia, Vol. 16, No. 10, 01.10.2002, p. 2046-2054.

Research output: Contribution to journalArticle

Todisco, E. ; Gaipa, G. ; Biagi, E. ; Bonamino, M. ; Gramigna, R. ; Introna, M. ; Biondi, A. / CD40 ligand-stimulated B cell precursor leukemic cells elicit interferon-γ production by autologous bone marrow T cells in childhood acute lymphoblastic leukemia. In: Leukemia. 2002 ; Vol. 16, No. 10. pp. 2046-2054.
@article{74aab66f0cdd41e4af42402d4767d813,
title = "CD40 ligand-stimulated B cell precursor leukemic cells elicit interferon-γ production by autologous bone marrow T cells in childhood acute lymphoblastic leukemia",
abstract = "Childhood B cell precursor acute lymphoblastic leukemia (BCP-ALL) cells, collected from bone marrow (BM) at diagnosis, were cultured, after thawing, on allogeneic human bone marrow stroma (HBMS) for 48 h in the presence of a soluble trimeric CD40 ligand (stCD40L) molecule. HBMS maintained leukemic cells viability in all tested cases (mean viability 85{\%}). Under these culture conditions we noticed upregulation or de novo expression of costimulatory molecules CD40, CD80 (B7-1) and CD86 (B7-2) in 22/22, 15/23 and 21/23 cases, respectively. Upregulation, in terms of fluorescence intensity, was also observed in the expression of MHC I, MHC II, CD54 (ICAM 1) and CD58 (LFA 3) molecules. HBMS alone, although to a lesser extent, was able to induce modulation of these molecules, but not CD80, in a similar proportion of cases. Neither stCD40L nor HBMS induced modulation of CD10 and CD34 molecules. Moreover, in 4/4 tested cases, stCD40L-stimulated ALL cells were able to induce allogeneic T cells proliferation. To evaluate whether leukemia-reactive T cells were detectable in the BM of ALL patients at diagnosis, stCD40L-stimulated ALL cells were co-cultured with autologous T cells (ratio 1:1), isolated from BM at diagnosis, for 4 days and a 24 h ELISPOT assay was applied to detect the presence of interferon-gamma (IFN-γ)-producing cells. In four of seven cases IFN-γ-producing cells were detected with frequencies of 1/900, 1/1560, 1/2150 and 1/1575 autologous T cells. These data confirm that stCD40L exposure can activate the antigen-presenting cell (APC) capacity of BCP-ALL cells cultured on HBMS and that ELISPOT assay can be used to measure the frequency of leukemia-reactive autologous T cells in the BM of ALL patients even after short-term culture with stCD40L-stimulated ALL cells.",
keywords = "CD40 ligand, Childhood ALL, ELISPOT, Immunology, Interferongamma, Stroma",
author = "E. Todisco and G. Gaipa and E. Biagi and M. Bonamino and R. Gramigna and M. Introna and A. Biondi",
year = "2002",
month = "10",
day = "1",
doi = "10.1038/sj.leu.2402672",
language = "English",
volume = "16",
pages = "2046--2054",
journal = "Leukemia",
issn = "0887-6924",
publisher = "Nature Publishing Group",
number = "10",

}

TY - JOUR

T1 - CD40 ligand-stimulated B cell precursor leukemic cells elicit interferon-γ production by autologous bone marrow T cells in childhood acute lymphoblastic leukemia

AU - Todisco, E.

AU - Gaipa, G.

AU - Biagi, E.

AU - Bonamino, M.

AU - Gramigna, R.

AU - Introna, M.

AU - Biondi, A.

PY - 2002/10/1

Y1 - 2002/10/1

N2 - Childhood B cell precursor acute lymphoblastic leukemia (BCP-ALL) cells, collected from bone marrow (BM) at diagnosis, were cultured, after thawing, on allogeneic human bone marrow stroma (HBMS) for 48 h in the presence of a soluble trimeric CD40 ligand (stCD40L) molecule. HBMS maintained leukemic cells viability in all tested cases (mean viability 85%). Under these culture conditions we noticed upregulation or de novo expression of costimulatory molecules CD40, CD80 (B7-1) and CD86 (B7-2) in 22/22, 15/23 and 21/23 cases, respectively. Upregulation, in terms of fluorescence intensity, was also observed in the expression of MHC I, MHC II, CD54 (ICAM 1) and CD58 (LFA 3) molecules. HBMS alone, although to a lesser extent, was able to induce modulation of these molecules, but not CD80, in a similar proportion of cases. Neither stCD40L nor HBMS induced modulation of CD10 and CD34 molecules. Moreover, in 4/4 tested cases, stCD40L-stimulated ALL cells were able to induce allogeneic T cells proliferation. To evaluate whether leukemia-reactive T cells were detectable in the BM of ALL patients at diagnosis, stCD40L-stimulated ALL cells were co-cultured with autologous T cells (ratio 1:1), isolated from BM at diagnosis, for 4 days and a 24 h ELISPOT assay was applied to detect the presence of interferon-gamma (IFN-γ)-producing cells. In four of seven cases IFN-γ-producing cells were detected with frequencies of 1/900, 1/1560, 1/2150 and 1/1575 autologous T cells. These data confirm that stCD40L exposure can activate the antigen-presenting cell (APC) capacity of BCP-ALL cells cultured on HBMS and that ELISPOT assay can be used to measure the frequency of leukemia-reactive autologous T cells in the BM of ALL patients even after short-term culture with stCD40L-stimulated ALL cells.

AB - Childhood B cell precursor acute lymphoblastic leukemia (BCP-ALL) cells, collected from bone marrow (BM) at diagnosis, were cultured, after thawing, on allogeneic human bone marrow stroma (HBMS) for 48 h in the presence of a soluble trimeric CD40 ligand (stCD40L) molecule. HBMS maintained leukemic cells viability in all tested cases (mean viability 85%). Under these culture conditions we noticed upregulation or de novo expression of costimulatory molecules CD40, CD80 (B7-1) and CD86 (B7-2) in 22/22, 15/23 and 21/23 cases, respectively. Upregulation, in terms of fluorescence intensity, was also observed in the expression of MHC I, MHC II, CD54 (ICAM 1) and CD58 (LFA 3) molecules. HBMS alone, although to a lesser extent, was able to induce modulation of these molecules, but not CD80, in a similar proportion of cases. Neither stCD40L nor HBMS induced modulation of CD10 and CD34 molecules. Moreover, in 4/4 tested cases, stCD40L-stimulated ALL cells were able to induce allogeneic T cells proliferation. To evaluate whether leukemia-reactive T cells were detectable in the BM of ALL patients at diagnosis, stCD40L-stimulated ALL cells were co-cultured with autologous T cells (ratio 1:1), isolated from BM at diagnosis, for 4 days and a 24 h ELISPOT assay was applied to detect the presence of interferon-gamma (IFN-γ)-producing cells. In four of seven cases IFN-γ-producing cells were detected with frequencies of 1/900, 1/1560, 1/2150 and 1/1575 autologous T cells. These data confirm that stCD40L exposure can activate the antigen-presenting cell (APC) capacity of BCP-ALL cells cultured on HBMS and that ELISPOT assay can be used to measure the frequency of leukemia-reactive autologous T cells in the BM of ALL patients even after short-term culture with stCD40L-stimulated ALL cells.

KW - CD40 ligand

KW - Childhood ALL

KW - ELISPOT

KW - Immunology

KW - Interferongamma

KW - Stroma

UR - http://www.scopus.com/inward/record.url?scp=0036800028&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036800028&partnerID=8YFLogxK

U2 - 10.1038/sj.leu.2402672

DO - 10.1038/sj.leu.2402672

M3 - Article

C2 - 12357356

AN - SCOPUS:0036800028

VL - 16

SP - 2046

EP - 2054

JO - Leukemia

JF - Leukemia

SN - 0887-6924

IS - 10

ER -