To explore the effect of cytokine therapy on the NADPH oxidase in mature myeloid cells, we isolated neutrophils from patients receiving recombinant human granulocyte colony stimulating factor (G-CSF) and recombinant human stem cell factor (SCF) and evaluated oxidase activity. All patients had relapsed neoplastic disease and were at least 3 three weeks since the last course of chemotherapy or cytokine therapy.
Methods
Stimulus induced superoxide anion (O2) production in response to PMA (200 ng/mL), fMLP (1 µmol/L), platelet activating factor (PAF, 2 µmol/L) priming of the fMLP induced response, and opsonized zymosan OZ (1 mg/mL) was measured. Polymorphonuclear leukocyte (PMN) subcellular components were prepared, after nitrogen cavitation, by separation on discontinuous sucrose gradients and NADPH oxidase activity was assessed in an SDS cell-free system.
Results
SCF had no effect on the activity of the neutrophil oxidase. Neutrophils isolated from patients treated with G-CSF and stimulated with PMA produced less (superoxide anion) O2 after therapy. PAF priming of the fMLP induced respiratory burst was also reduced after therapy with G-CSF. Subcellular NADPH oxidase activity was reduced before cytokine therapy commenced. This activity did not improve with cytokine treatment.
Conclusions
It appears likely from this study that G-CSF therapy, with or without SCF, does not cause significant enhancement of neutrophil NADPH oxidase activity.
NicolaNA. Granulocyte colony stimulating factor. In: DexterTM, GarlandJM, TestaNG, eds. Colony-stimulating factors. New York:Marcel Dekker, Inc. 1990:77–109.
2.
GlaspyJA, BleeckerG, CrawfordJ, StollerR, StraussM. The impact of therapy with filgrastim (recombinant granulocyte colony-stimulating factor) on the health care costs associated with cancer chemotherapy. Eur J Cancer1993;29A(7 Suppl):S23–30.
3.
LinchDC, ScarffeH, ProctorS, ChopraR, TaylorPR, Morgen-sternG, CunninghamD, BurnettAK, CawleyJC, FranklinIM. Randomised vehicle-controlled dose-finding study of glycosylated recombinant human granulocyte colony-stimulating factor after bone marrow transplantation. Bone Marrow Transplant1993;11:307–11.
4.
BalazovichKJ, AlmeidaHI, BoxerLA. Recombinant human G-CSF and GM-CSF prime human neutrophils for superoxide production through different signal transduction mechanisms. J Lab Clin Med1991;118:576–84.
5.
YuoA, KitagawaS, OhsakaA, OhtaM, MiyazonoK, OkabeT, UrabeA, SaitoM, TakakuF. Recombinant human granulocyte colony-stimulating factor as an activator of human granulocytes: potentiation of responses triggered by receptor-mediated agonists and stimulation of C3bi receptor expression and adherence. Blood1989;74:2144–9.
6.
FossatC, StoppaAM, SaintyD, BlaiseD, ViensP, Juhan-VagueI, MaraninchiD, CarcassonneY. G-CSF in vivo improves neutrophils bactericidal activity against pseudomonas aeruginosae in cancer patients [abstract]. Blood1990;76:144A.
7.
KaposztaR, MarodiL. Chronic neutropenia and defect in superoxide generation of granulocytes in two patients: enhancement of bactericidal capacity and respiratory burst activity by treatment with recombinant human granulocyte colony-stimulating factor. Pediatr Res1995;37:50–5.
8.
LilesWC, HuangJE, van BurikJH, BowdenRA, DaleDC. Granulocyte colony stimulating factor administered in vivo augments neutrophil-mediated activity against opportunistic fungal pathogens. J Infect Dis1997;175:1012–5.
CairoMS, PlunkettJM, NguyenA, van de VenC. Effect of stem cell factor with and without granulocyte colony-stimulating factor on neonatal hematopoiesis: in vivo induction of newborn myelopoiesis and reduction of mortality during experimental group B streptococcal sepsis. Blood1992;80:96–101.
11.
SteinshamnS, BerghK, WaageA. Effects of stem cell factor and granulocyte colony-stimulating factor on granulocyte recovery and Candida albicans infection in granulocytopenic mice. J Infect Dis1993;168:1444–8.
AmbrusoDR, BrentwoodB, HensonPM, JohnstonRBJr.. Oxidative metabolism of cord blood neutrophils: relationship to content and degranulation of cytoplasmic granules. Pediatr Res1984;18:1148–53.
14.
RosenthalJ, ThurmanGW, CusackN, PetersonVM, MalechHL, AmbrusoDR. Neutrophils from patients after burn injury express a deficiency of the oxidase components p47-phox and p67-phox. Blood1996;88:4321–9.
15.
AmbrusoDR, BolscherBG, StokmanPM, VerhoevenAJ, RoosD. Assembly and activation of the NADPH:O2 oxidoreductase in human neutrophils after stimulation with phorbol myristate acetate. J Biol Chem1990;265:924–30.
16.
AmbrusoDR, StorkLC, GibsonBE, ThurmanGW. Increased activity of the respiratory burst in cord blood neutrophils: kinetics of the NADPH oxidase enzyme system in subcellular fractions. Pediatr Res1987;21:205–10.
17.
ColeJW, GrizzleJE. Applications of multivariate analysis of variance to repeated measurements experiments. Biometrics1966;22:810–28.
18.
SchrotenH, RoeslerJ, BreidenbachT, WendelD, ElsnerJ, SchweitzerS, ZeidlerC, BurdachS, Lohmann-MatthesM, WahnV. Granulocyte and granulocyte-macrophage colony-stimulating factors for treatment of neutropenia in glycogen storage disease type lb. J Pediatr1991;119:748–54.
19.
NegrinRS, HaeuberDH, NaglerA, OldsLC, DonlonT, SouzaLM, GreenbergPL. Treatment of myelodysplastic syndromes with recombinant human granulocyte colony-stimulating factor. Ann Intern Med1989;110:976–84.
20.
McCawleyU, KorchakHM, DouglasSD, CampbellDE, ThorntonPS, StanleyCA, BakerL, KilpatrickL. In vitro and in vivo effects of granulocyte colony-stimulating factor on neutrophils in glycogen storage disease type IB: granulocyte colony-stimulating factor therapy corrects the neutropenia and the defects in respiratory burst activity and Ca2+ mobilization. Pediatr Res1994;35:84–90.
21.
OhsakaA, KitagawaS, YuoA, MotoyoshiK, OhtaM, MiuraY, TakakuF, SaitoM. Increased respiratory burst activity of neutrophils in patients with aplastic anemia: effects of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. Exp Hematol1992;20:1090–3.
22.
McNieceIK, BriddellRA, HartleyCA, AndrewsRG. The role of stem cell factor in mobilization of peripheral blood progenitor cells: synergy with G-CSF. Stem Cells (Dayt)1993;(11Suppl):83–8.
23.
ChattaGS, PriceTH, AllenRC, DaleDC. Effects of in vivo recombinant methionyl human granulocyte colony- stimulating factor on the neutrophil response and peripheral blood colony-forming cells in healthy young and elderly adult volunteers. Blood1994;84:2923–9.
24.
MaceyMG, SangsterJ, KelseySM, NewlandAC. Pilot study: effects of G-CSF on neutrophil ex-vivo function post bone marrow transplantation. Clin Lab Haematol1993;15:79–85.
25.
DaleDC, LilesWC, SummerWR, NelsonS. Review: granulocyte colony-stimulating factor-role and relationships in infectious diseases. J Infect Dis1995;172:1061–75.
26.
ShearmanMS, HeyworthCM, DexterTM, HaefnerB, OwenPJ, WhettonAD. Haemopoietic stem cell development to neutrophils is associated with subcellular redistribution and differential expression of protein kinase C subspecies. J Cell Sci1993;104:173–80.
27.
OhsakaA, KitagawaS, SakamotoS, MiuraY, TakanashiN, TakakuF, SaitoM. In vivo activation of human neutrophil functions by administration of recombinant human granulocyte colony-stimulating factor in patients with malignant lymphoma. Blood1989;74:2743–8.
28.
TreweekeAT, AzizKA, ZuzelM. The role of G-CSF in mature neutrophil function is not related to GM-CSF-type cell priming. J Leukoc Biol1994;55:612–6.
29.
KhwajaA, CarverJE, LinchDC. Interactions of granulocyte macrophage colony-stimulating factor (CSF), granulocyte CSF, and tumor necrosis factor alpha in the priming of the neutrophil respiratory burst. Blood1992;79:745–53.
30.
AmbrusoDR, PetersonVP. Postburn defects in phagocyte function. In: Phagocyte production and function following burn injury. Austin:R.G. Landes Company, 1994:93–124.
31.
CioffiWG, BurlesonDG, PruittBAJr. Leukocyte responses to injury. Arch Surg1993;128:1260–7.
32.
ChenTP, RobertsRL, WuKG, AnkBJ, StiehmER. Decreased superoxide anion and hydrogen peroxide production by neutrophils and monocytes in human immunodeficiency virus-infected children and adults. Pediatr Res1993;34:544–50.
33.
LejeuneM, SaribanE, CantinieauxB, FersterA, DevalckC, FonduP. Granulocyte functions in children with cancer are differentially sensitive to the toxic effect of chemotherapy. Pediatr Res1996;39:835–42.
34.
LeaveyPJ, ThurmanG, AmbrusoDR. The effect of in vivo granulocyte colony stimulating factor (G-CSF) administration on the NADPH oxidase from peripheral blood neutrophils [abstract]. J Invest Med1996;44:277A.
