Constitutive and modulated cytokine expression in two permanent human bone marrow stromal cell lines.
We present a detailed analysis of cytokine expression patterns of the two permanent human bone marrow stromal cell lines, L87/4 and L88/5. These cell lines, previously established in our laboratory, are highly radiotolerant without cell detachment and support long-term cultures of CD(34+)-enriched human cord blood cells. RT-PCR analysis of 22 different cytokines or cytokine receptor mRNAs showed an almost identical expression pattern in the two stromal cell lines compared to primary human Dexter-type stroma. Since stromal feeder lines employed in long-term cultures usually are irradiated and grown in media containing corticosteroids, we analyzed the impact of irradiation and dexamethasone on cytokine production in the two cell lines by RT-PCR, Northern blot analysis, bioassays, and RIAs. By RT-PCR analysis, constitutive mRNA expression of c-kit, G-CSF, GM-CSF, IL-1 beta, IL-6, IL-7, IL-8, IL-11, Kit ligand (KL), LIF, M-CSF, MIP-1 alpha, TGF-beta, and TNF-alpha was demonstrated in both cell lines, with L87/4 a more potent cytokine producer than L88/5. Northern blot data showed an increase in mRNA levels for GM-CSF, IL-1 beta, and LIF by irradiation and IL-1 alpha treatment in both cell lines. IL-1 alpha-induced GM-CSF, IL-1 beta, IL-6, IL-11, and LIF mRNA levels were reduced by the addition of dexamethasone, whereas dexamethasone had no influence on the amounts of IL-1 alpha-induced G-CSF mRNA. L87/4 and, to a lower extent, L88/5 cells showed dexamethasone-dependent increases in KL mRNA, while KL mRNA levels were not stimulated by IL-1 alpha.
GSF-Institut für Experimentelle Hämatologie, München, Germany.
K
K
Thalmeier
P
P
Meissner
G
G
Reisbach
L
L
Hültner
B T
BT
Mortensen
A
A
Brechtel
R A
RA
Oostendorp
P
P
Dörmer
D001483:Base Sequence; D015152:Blotting, Northern; D001853:Bone Marrow / Q000378:metabolism* / Q000528:radiation effects; D001854:Bone Marrow Cells; D002460:Cell Line; D016207:Cytokines / Q000235:genetics*; D003907:Dexamethasone / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects / Q000528:radiation effects; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D006131:Growth Inhibitors / Q000235:genetics; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics / Q000494:pharmacology; D017370:Interleukin-11 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase; D017154:Stromal Cells / Q000378:metabolism* / Q000528:radiation effects
D016207:Cytokines; D006131:Growth Inhibitors; D007375:Interleukin-1; D017370:Interleukin-11; D015850:Interleukin-6; C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines; D012333:RNA, Messenger; D003907:Dexamethasone; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D012194:RNA-Directed DNA Polymerase