Amplification of T cells from human cord blood in serum-deprived culture stimulated with stem cell factor, interleukin-7 and interleukin-2.
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 V(beta)-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.
Sanchez
M
M
Laboratory of Cell Biology, Istituto Superiore di Sanita, Rome, Italy.
Alfani
E
E
Migliaccio
A R
AR
Bonfini
T
T
Migliaccio
G
G
D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division / Q000187:drug effects; D016895:Culture Media, Serum-Free; D036861:Delivery, Obstetric; D005312:Fetal Blood / Q000276:immunology*; D005434:Flow Cytometry; D019650:Hematopoietic Stem Cell Mobilization / Q000379:methods; D006801:Humans; D016130:Immunophenotyping; D007231:Infant, Newborn; D007376:Interleukin-2 / Q000494:pharmacology*; D015851:Interleukin-7 / Q000494:pharmacology*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019089:Stem Cell Factor / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D013601:T-Lymphocytes / Q000276:immunology*
D016895:Culture Media, Serum-Free; D007376:Interleukin-2; D015851:Interleukin-7; D016693:Receptors, Antigen, T-Cell, alpha-beta; D019089:Stem Cell Factor