TY - JOUR
T1 - Amplification of T cells from human cord blood in serum-deprived culture stimulated with stem cell factor, interleukin-7 and interleukin-2
AU - Sanchez, M.
AU - Alfani, E.
AU - Migliaccio, A. R.
AU - Bonfini, T.
AU - Migliaccio, G.
N1 - Funding Information:
We thank Dr M Marceca (II Clinica Ostetrica, Università agli Studi ‘La Sapienza’, Rome, Italy) for providing CB samples. This work was supported by institutional funds of Istituto Superiore di Sanita’, Progetto Finalizzato 1%, Ricerca Corrente and Progetti di Ricerca di Interesse Nazionale 2000, from the Ministry of Health; Progetto Strategico Oncologia CNR-MIUR legge 449/99 and Grant no. E1172 from the Telethon Foundation.
PY - 2003/4
Y1 - 2003/4
N2 - We report the effects exerted by cytokine combinations, including stem cell factor (SCF), interleukin-7, interleukin-4 and interleukin-2, on the amplification of T cells from cord blood (CB) mononuclear cells cultured for 10-11 days under serum-deprived conditions. Of all the combinations investigated, SCF + interleukin-7 sustained the best fold increase (FI) of total nucleated cells (FI = 6.4±1.17), amplifying preferentially CD4+ over CD8+ T-cell subsets (FI = 4.72±0.79 vs 2.73±1.2, respectively, P < 0.05). The addition of interleukin-2 to this combination did not significantly increase the total number of cells generated (FI = 7.4±2.27), but allowed preferential amplification of CD8+ over CD4+ T cells (FI = 6.04±0.14 vs 1.67±0.6, respectively, P < 0.05). Single-strand conformation polymorphism analysis of the T-cell receptor Vβ-chain rearrangements expressed by the expanded T cells indicated that the complexity of the T-cell repertoire had increased after 10 days of culture in the presence of SCF and IL-7. Interestingly, a modest expansion (FI = 8.67±1.5 of myeloid progenitor cells was also observed in these cultures. These results indicate that it is possible to expand specific T-cell subsets for adoptive immunotherapy without losing myeloid progenitor cells necessary for neutrophil recovery after CB transplantation, by modulating the cytokines added to the cultures.
AB - We report the effects exerted by cytokine combinations, including stem cell factor (SCF), interleukin-7, interleukin-4 and interleukin-2, on the amplification of T cells from cord blood (CB) mononuclear cells cultured for 10-11 days under serum-deprived conditions. Of all the combinations investigated, SCF + interleukin-7 sustained the best fold increase (FI) of total nucleated cells (FI = 6.4±1.17), amplifying preferentially CD4+ over CD8+ T-cell subsets (FI = 4.72±0.79 vs 2.73±1.2, respectively, P < 0.05). The addition of interleukin-2 to this combination did not significantly increase the total number of cells generated (FI = 7.4±2.27), but allowed preferential amplification of CD8+ over CD4+ T cells (FI = 6.04±0.14 vs 1.67±0.6, respectively, P < 0.05). Single-strand conformation polymorphism analysis of the T-cell receptor Vβ-chain rearrangements expressed by the expanded T cells indicated that the complexity of the T-cell repertoire had increased after 10 days of culture in the presence of SCF and IL-7. Interestingly, a modest expansion (FI = 8.67±1.5 of myeloid progenitor cells was also observed in these cultures. These results indicate that it is possible to expand specific T-cell subsets for adoptive immunotherapy without losing myeloid progenitor cells necessary for neutrophil recovery after CB transplantation, by modulating the cytokines added to the cultures.
KW - Cord blood
KW - Ex vivo amplification
KW - Interleukin-2
KW - Interleukin-7
KW - Serum-free cultures
KW - Stem cell factor
KW - T lymphocytes
UR - http://www.scopus.com/inward/record.url?scp=0038702467&partnerID=8YFLogxK
U2 - 10.1038/sj.bmt.1703904
DO - 10.1038/sj.bmt.1703904
M3 - Article
C2 - 12692612
AN - SCOPUS:0038702467
SN - 0268-3369
VL - 31
SP - 713
EP - 723
JO - Bone Marrow Transplantation
JF - Bone Marrow Transplantation
IS - 8
ER -