Maintenance and characterization of lymphocytes in human long term bone marrow cultures to study immunotoxicity.
Immunotoxicity of xenobiotics is of growing concern for various levels in society, including industry and regulatory authorities. Despite that EU legislation aims at reducing the number of laboratory animals by promoting the development of alternative validated methods, at present, immunotoxicity is generally evaluated through standard in vivo toxicity studies. The lack of alternative methods is due, in particular, to the complexity of the immune system and its responses, but possibly alternative methods for immunosuppressive chemicals are most achievable. The present study describes a long term culture (LTC) method capable of inducing the formation of lymphocyte subsets from human mononuclear bone marrow cells that may allow evaluation of lymphotoxicity. The LTC consisted of a two stages: a myeloid stage to allow the formation of a stromal layer and a lymphoid stage to allow expansion of lymphocytes. Results show that the use of IL-7 in LTC inhibits precursor and mature B cells, while it supports the proliferation of CD3(+)CD8(+) and CD3(+)CD4(+) T-cells. The bone marrow LTC model may in future be used to test the effect of xenobiotics on stromal dependent lymphocyte formation.
Carfí
M
M
ECVAM, IHCP, JRC-TP 580-via E Fermi 2749, 21027 Ispra, Italy. marycarfi@hotmail.com
Bowe
G
G
Ferrario
D
D
Pieters
R
R
Gribaldo
L
L
D018941:Antigens, CD19 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D015704:CD4 Antigens / Q000378:metabolism; D016827:CD8 Antigens / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D006801:Humans; D015851:Interleukin-7 / Q000494:pharmacology; D008214:Lymphocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D018675:Toxicity Tests; D015262:Xenobiotics / Q000633:toxicity
D018941:Antigens, CD19; D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D015851:Interleukin-7; D015262:Xenobiotics