Increased expansion of V alpha 24+ T cells derived from G-CSF-mobilized peripheral blood stem cells as compared to peripheral blood mononuclear cells following alpha-galactosylceramide stimulation.
In the present study, unpurified peripheral blood mononuclear cells (PBMCs) from various sources, including steady-state blood (normal donors) and granulocyte colony-stimulating factor (G-CSF)-mobilized blood (cancer patients and normal donors) (G-PBSC), were cultured in RPMI-1640 in the presence of IL-2 and alpha-galactosylceramide (alpha-GalCer) to expand V alpha 24(+) T cells, and their expansion kinetics were compared. G-CSF-mobilized cells showed markedly higher expansion potential (350-fold expansion of V alpha 24(+) T cells, regardless of whether the cells were from cancer patients or normal donors) than steady-state cells (15-fold expansion, compared to the initial inoculums) (n = 5, P < 0.01). We also confirmed that the CD14(-) fraction of G-PBSCs contained a large number of precursors of V alpha 24(+) T cells, compared to PBSCs, as well as a large number of CD14(+) cells, which assist V alpha 24(+) T cell proliferation. Our simple and practical procedure, which eliminates complicated cell manipulation (including cell purification), produces efficient expansion of V alpha 24(+) T cells when G-CSF-mobilized blood cells are cultured with alpha-GalCer.
Asada-Mikami
Rumiko
R
Pharmacology Division, National Cancer Center Research Institute and Hematopoietic Stem Cell Transplant/Immunotherapy Division, National Cancer Center Hospital, Chuo-ku, Tokyo 104-0045, Japan.
Heike
Yuji
Y
Harada
Yukie
Y
Kanai
Sachiyo
S
Ikarashi
Yoshinori
Y
Kato
Kazunori
K
Shirakawa
Kazuo
K
Takaue
Yoichi
Y
Abe
Tatsuo
T
Wakasugi
Hiro
H
D001854:Bone Marrow Cells; D002469:Cell Separation; D002478:Cells, Cultured; D005699:Galactosylceramides / Q000494:pharmacology*; D016179:Granulocyte Colony-Stimulating Factor / Q000378:metabolism* / Q000494:pharmacology*; D019650:Hematopoietic Stem Cell Mobilization; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000145:classification / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D016130:Immunophenotyping; D039564:Integrin alphaV / Q000502:physiology*; D007963:Leukocytes, Mononuclear; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D009369:Neoplasms / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D011994:Recombinant Proteins / Q000494:pharmacology; D013601:T-Lymphocytes / Q000378:metabolism*
D005699:Galactosylceramides; D039564:Integrin alphaV; D018950:Lipopolysaccharide Receptors; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor