Recombinant human granulocyte-macrophage colony-stimulating factor (rH GM-CSF) regulates f Met-Leu-Phe receptors on human neutrophils.
The regulation of mature human neutrophil function by recombinant human granulocyte-macrophage colony-stimulating factor (rH GM-CSF) was studied. Preincubation of neutrophils with this CSF did not stimulate superoxide anion directly but enhanced the subsequent release of superoxide anion in response to stimulation with the bacterial product formylmethionylleucyl-phenylalanine (f Met-Leu-Phe). Enhanced superoxide anion production was evident by 5 min and reached a plateau at 30 min. In contrast, neutrophils preincubated with rH GM-CSF exhibited reduced chemotaxis under agarose in response to a gradient of f Met-Leu-Phe. The inhibition of neutrophil migration was dependent on the dose of rH GM-CSF and exhibited a time-course similar to the effect on superoxide production. Binding studies of f Met-Leu-[3H]Phe to purified human neutrophils revealed heterogeneous binding to unstimulated cells. Two affinity components were identified. The high-affinity component consisted of approximately 2000 sites/cell and had an average Kd of 4 +/- 2 nM (n = 6). The low-affinity component consisted of approximately 40,000 sites/cell and had an average Kd of 220 +/- 130 nM (n = 6). rH GM-CSF caused conversion to a linear Scatchard plot showing no significant change in total binding sites but a single Kd of 30 +/- 10 nM. These data indicate that rH GM-CSF may influence neutrophil responses to f Met-Leu-Phe by regulating the affinity of f Met-Leu-Phe receptors.
Atkinson
Y H
YH
Division of Human Immunology, Institute of Medical and Veterinary Science, Adelaide, South Australia.
Lopez
A F
AF
Marasco
W A
WA
Lucas
C M
CM
Wong
G G
GG
Burns
G F
GF
Vadas
M A
MA
D002634:Chemotaxis, Leukocyte; D003115:Colony-Stimulating Factors / Q000494:pharmacology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism* / Q000502:physiology; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism
D003115:Colony-Stimulating Factors; D006133:Growth Substances; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor