The colony stimulating factors (CSF's) are haematopoietic growth factors that stimulate cell division and differentiation in blood cell precursors. There are at least ...
Biochemical Society Transactions ( I99 I ) I 9 Recombinant Human Granulocyte Colony Stimulating Factor (rhG-CSF) and superoxide production in a haematopoietic cell line. WILLIAM S. GILMORE, B.M. HANNIGAN, S.J. EASTON, C.P. MCGUCKIN and A. A. NELSON Biomedical Sciences Research Centre, University of Ulster a t Coleraine, Cromore Road, Coleraine, Co Londonderry, BT52 lSA, Northern Ireland, U.K. The colony stimulating factors (CSF's) are haematopoietic growth factors that stimulate cell division and differentiation in blood cell precursors. There are at least four CSFs involved in the proliferation of myeloid cells multi colony stimulating factor (multi-CSF), granulocytelmonocyte colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF) and monocyte colony stimulating factor (M-CSF).(l) In addition to stimulating the growth and differentiation of haematopoietic cells the CSF's enhance the biological functions of the mature cells (2-7). It has been reported that the CSFs are lineage specific in the enhancement of biological function of mature granulocytes and monocytes: GM-CSF enhancing both granulocytic and monocytic function and G - C S F enhancing granulocytes (2-9). In order to test the lineage specificity of GCSF we have investigated the ability of recombinant human granulocyte colony stimulating factor (rhG-CSF) t o enhance superoxide anion production in the monocytic cell line U-937. The methods have been described previously in detail (9). Briefly U-937 cells were washed and plated on 96 well plates at a concentration of 3-3.5~10' cells p e r well, containing cytochrome-c (final concentration 160uM) and with o r without stimulus of either G-CSF (2000 units/ml) or G-CSF (2000 units/ ml) and phorbol myristate acetate (PMA) at a final concentration of 500ng/ml. Control wells contained the enzyme superoxide dismutase (final concentration lSuM).(Total well volume was 0.17Oml). All t a u were done in replicates of six. The absorbances were read on a standard Microelisa reader at 550nm over a period of 90-100 minutes. The rhG-CSF was obtained from Amersham, England and all other reagents from Sigma, Poole, England. Table 1
Production of Superoxide by U937 cells.
Each value represents the mean f SD for six wells (corrected for control wells and cell number.) Stimulant
25.0
PMA PMA
Superoxide (nmoles/min/l@ cells)
+ rhG-CSF
f
0.542
48.5
2
0.518
T h e results a r e summarised in Table 1. It can be seen that superoxide production by the U-937 cells is enhanced by the addition of the rhG-CSF, suggesting that in addition to enhancing superoxide production in granulocytes, G-CSF may also act to enhance the biological function of monocytic cells. On the other hand the U-937 cell-line used may not truly reflect monocytic cell function. The results however indicate that G-CSF may act o n a number of different cell types.
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