Immunology 1992 77 147-149
BRIEF COMMUNICATION
Neutrophil priming by hepatocyte growth factor,
a
novel cytokine
W. JIANG, M. C. A. PUNTIS, T. NAKAMURA* & M. B. HALLETT University Department of Surgery, University of Wales College of Medicine, Cardiff, U.K. and *Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan
Accepted for publication 12 April 1992
SUMMARY We demonstrate here that the recently defined cytokine hepatocyte growth factor (HGF) 'primes' human neutrophils. Recombinant human HGF over the concentration range 0-1-20 ng/ml increased the neutrophil response to f-met-leu-phe by up to 200%, and required only a short preincubation, 10 min producing the maximum effect. Priming was independent of changes in cytosolic-free calcium homeostasis. We conclude that HGF may be a physiologically important cytokine with 'priming' activity for neutrophils.
Activation of the non-mitochondrial oxidase of neutrophils, by generating toxic metabolites of oxygen, plays a crucial role in combating infection"2 but also causes tissue damage.3'4 This oxidative response may be enhanced by the activity of neutrophil 'primers', such as the cytokines tumour necrosis factor-ca (TNF-cc),5 interleukin-1 (IL-1),6 IL-6,7 IL-8,8 and granulocytemacrophage colony-stimulating factor (GM-CSF).9 Hepatocyte growth factor (HGF) is a recently defined growth factor which is a heterodimer molecule composed of a 69,000 MW a-subunit and a 34,000 MW f-subunit. This factor was first found in sera from partially hepatectomized rats,'0 its complementary DNA (cDNA) has recently been cloned from both the rat" and man.'2 HGF provokes DNA synthesis in hepatocytes in primary culture'3 and acts via a specific receptor on hepatocytes."4 HGF also affects keratinocytes, melanocytes and other cells.'5-'7 HGF has been reported to appear in the sera and other biological fluids in some diseases. 18-20 Recent reports have shown that HGF is identical to scatter factor which can dissociate epithelial cells.2' As HGF shares with other neutrophil primers both growth factor-like activity and a monocytic origin (from Kupffer cells),22 we have investigated whether HGF has priming activity for the neutrophil oxidase response. Human neutrophils were isolated from normal blood by dextran sedimentation and Ficoll-Hypaque centrifugation, and then resuspended in HEPES-buffered Krebs medium. The production of reactive oxygen metabolites was monitored using luminol-dependent chemiluminescence as reported previously.23 The cytosolic free Ca2+ was measured using dual wavelength fluorometry of Fura-2 (Spex CM, Instrument SA, Stanmore,
U.K.), as previously described.24 The cytosolic Fura-2 concentration of approximately 100 yM increased the cytosolic Ca2+ buffering capacity by 10-20%. Recombinant human hepatocyte growth factor was over 90% pure, half maximal activity on hepatocyte DNA synthesis was 3-75 ng/ml."I Stock solutions of 1 ug/ml were kept at -70° and diluted immediately before use. In order to determine the responsiveness of neutrophils to other 'primers', human recombinant IL-8 (89/520, NBSB, Potters Bar, U.K.) and TNF-a (a gift from Dr N. Matthews, UWCM, Cardiff, U.K.) were also used. Fura-2/AM was purchased from Molecular Probes (OR). All other materials were purchased from Sigma (Poole, Dorset, U.K.) unless otherwise stated. Priming occurred over a wide range of rHGF concentrations. Maximal priming was achieved by 10 ng/ml which enhanced the response by over 200%. The lowest detectable
240
C 190
0S t
140 ,
I
001
0-1
1
10
100
rHGF (ng/ml) Figure 1. The dose response of HGF priming on neutrophils. Neutrophils were preincubated for 10 min at 370 with rHGF. At the end of this period the luminol-dependent chemiluminescence rate was recorded before (0) and at the peak rate after stimulation (0) with FMLP (1 pM) plus cytochalasin B (5 pg/ml). The results are expressed as per cent priming, calculated as percentage of the response in the absence of rHGF.
Abbreviations: FMLP, f-met-leu-phe; GM-CSF, granulocytemacrophage colony-stimulating factor; HGF, hepatocyte growth factor; IFN-y, interferon-y; IL, interleukin; rHGF, recombinant hepatocyte growth factor; TNF-a, tumour necrosis factor-a. Correspondence: Dr W. Jiang, University Dept. of Surgery, University of Wales College of Medicine, Cardiff, U.K. :
147
148
W. Jiang et al. 800
800 r
Control
600
600
rHGF (50 ng/ml)
1 yilZ^q 1/ 5/il
Qb 200
0
Medium
FMLP
200
o*
rHFML
0 20 40 60 80 100 120 140 160 180 200 220 240 Time (second) Figure 2. Cytosolic-free Ca2 + changes in neutrophils. HGF was tested at concentrations 1, 10, 20, 30 and 50 ng/ml. No cytosolic-free calcium change was observed at these concentrations. At the first arrow either control medium or HGF (50 ng/ml) was added to the cell suspension. At the second arrow in each experiment FMLP (1 yM) was added.
0 20 40 60 80 100 120 140 160 180 200 220 240
enhancement occurred at 01 ng/ml (Fig. 1). In five separate experiments results show that the effect of preincubation with rHGF (5 ng/ml) for 10 min at 370, was to increase the production of reactive oxygen metabolites induced by the peptide f-met-leu-phe (FMLP, I tiM) plus cytochalasin B (5 jug/ ml) by 145 + 13-0% (P < 0-05 by Mann-Whitney U-test). This was comparable with IL-8 (2-0 ng/ml) and TNF-a (1 0 ng/ml) which prime neutrophils by 185-2+ 17-1 % and 136-4+ 19-4% respectively. Maximum priming by rHGF occurred rapidly, within 10 min, and remained at this level for up to 90 min (data not shown), suggesting that a protein synthesis step was not involved. If FMLP was absent no direct activation of the neutrophil oxidase was observed using concentrations of rHGF alone up to 20 ng/ml. It was unlikely that the effects reported here on neutrophils could be attributed to contaminating lipopolysaccharide (LPS) because heat (800, 30 min) abolished the priming effect (101 1-% after heat treatment of HGF) whereas polymixin B (1 mg/ml), which binds LPS, had no effects (149-0% in the presence of HGF and polymixin B). Furthermore, the priming effects were completely blocked by a specific rabbit anti-human HGF antibody (100-2% in the presence of HGF with anti-HGF). The effect on neutrophils was therefore attributed to rHGF. The possibility that priming by rHGF was mediated by a change in Ca2+ homoeostasis within the neutrophil was examined by measurement of the cytosolic-free Ca2+ concentration in neutrophils during incubation with HGF and during subsequent activation with FMLP. At concentrations up to 50 ng/ ml, HGF had no detectable effect on cytosolic-free Ca2 . Furthermore there was no detectable effect of rHGF on the peak, timing or kinetics of the transient cytosolic free Ca2+ rise induced by FMLP (Fig. 2). This eliminates the possibility that the mechanism of the activity of HGF was via up-regulation of FMLP receptors. The results of the experiments described here demonstrate a novel and previously undescribed property of HGF, namely the ability to prime the neutrophil oxidase response. HGF is a recently defined growth factor, synthesized by liver macrophages (Kupffer cells), endothelial cells and also expressed on
certain other tissues such as brain, lung, thyroid and pancreas.2225 It has major effects on hepatocytes, increasing DNA synthesis. We propose that HGF should, in addition, be added to the list of physiologically relevant neutrophil primers, which includes TNF-x, IL-1, IL-6, IL-8, GM-CSF, TGF-f3, IFN-y and substance P. It should be noted that although hepatocyte growth factor receptors have been detected on several cell and tissue types, such as hepatocytes, lung, kidney, spleen and most epithelial cells,'4 there are no reports at present which demonstrate their presence on neutrophils. Although the mechanisms by which HGF act are not yet established, Rubin et al.26 have recently reported that rHGF induces tyrosine phosphorylation in epithelial cells and melanocytes, and have suggested that the signalling pathway for this factor may involve the activation of tyrosine kinase. The possibility exists that HGF may act similarly in neutrophils and interact synergistically with FMLP-induced increased phosphotyrosine levels.27 HGF produces priming with a similar magnitude and a similar lack of effect on cytosolic-free Ca2+ homoeostasis as TNF-a, suggesting that these primers may share a common mechanism.2829 Further study will be required to establish this.
ACKNOWLEDGMENTS JWG thanks Scotia Pharmaceutical Ltd and MBH thanks the Arthritis and Rheumatism Council (U.K.) for support.
REFERENCES 1. FERRANTE A. (1989) Tumor necrosis factor alpha potentiates neutrophil antimicrobial activity: increased fungicidal activity against Torulopsis glabrata and Candida albicans and associated increases in oxygen radical production and lysosomal enzyme release. Infect. Immun. 57, 2115. 2. KUMARATILAKE L. M., FERRANTEA. & RZEPCZYK C. (199 1) The role of T lymphocytes in immunity to Plasmodiumfalciparum. Enhancement of neutrophil-mediated parasitic killing by lymphotoxin and IFN-gamma: comparisons with tumor necrosis factor effects. J. Immunol. 146, 762.
Neutrophil priming by hepatocyte growth factors 3. TELLADO J.M. & CHRISTOU V. (1991) Critically ill anergic patients demonstrate polymorphonuclear neutrophil activation in the intravascular compartment with decreased cell delivery to inflammatory focci. J. Leukoc. Biol. 50, 547. 4. KOWANKO I.C. & FERRANTE A. (1991) Granulocyte-macrophage colony-stimulating factor auguments neutrophil-mediated cartilage degradation and neutrophil adherence. Arthritis Rheum. 34, 1453. 5. KLEBANOFF S.J., VADAS M.M., HARLAN J.M., SPARKS L.H., GAMBLE J.R., AGoSTI J.M. & WALTERSDORPH A.M. (1986) Stimulation of neutrophils by tumor necrosis factor. J. Immunol. 136, 420. 6. FERRANTE A., NANDOSKAR M., WALZ A., GOH D.H.B. & KOWANKO I.C. (1988) Effects of tumour necrosis factor alpha and interleukin- l alpha and beta on human neutrophil migration, respiratory burst and degranulation. Int. Archs Allergy appl. Immunol. 86, 82. 7. BORISH L., ROSENBAUM R., ALBURY L. & CLARK S. (1989) Activation of neutrophils by recombinant interleukin-6. Cell. Immunol. 121, 280. 8. BAGGIOLINI M., WALZ A. & KUNKEL S.L. (1989) Neutrophilactivating peptide-1/interleukin-8, a novel cytokine that activates neutrophils. J. clin Invest 84, 1045. 9. MEGE J.L., GOMEZ-CAMBRONERO J., MOLSKi T.F.P., BECKER E.L. & SHA'AFI R.I. (1989) Effect of granulocyte-macrophage colonystimulating factor on superoxide production in cytoplasts and intact human neutrophils. J. Leukoc. Biol. 46, 161. 10. NAKAMURA T., NAWA K. & ICHIHARA A. (1984) Partial purification and characterization of hepatocyte growth factor from serum of hepatectomized rats. Biochem. biophys. Res. Commun. 122, 1450. 11. NAKAMURA T., NISHIZAWA T., HAGIYA M., SEKI T., SHIMONISHI M., SUGIMURA A., TASHIRO K. & SHIMIZU S. (1989) Molecular cloning and expression of human hepatocyte growth factor. Nature,
342,440. 12. TASHIRo K., HAGIYA M., NISHIZAWA T., SEKI T., SHIMONISHI M., SHIMIZU S. & NAKAMURA T. (1990) Deduced primary structure of rat hepatocyte growth factor and expression of the mRNA in rat tissues. Proc. natl. Acad. Sci. U.S.A. 87, 3200. 13. ZARNEGAR R. & MICHALOPOULOS G. (1989) Purification and biological characterization of human hepatopoietin A, a polypeptide growth factor for hepatocytes. Cancer Res. 49, 3314. 14. ZARNEGAR R., DEFRANCES M.C., OLIVE L. & MICHALOPOULOS G. (1990) Identification and partial characterization of receptor binding sites for HGF on rat hepatocytes. Biochem. biophys. Res. Commun. 173, 1179. 15. KAN M.K., ZHANG G., ZARNEGAR R., MICHALOPOULOS G., MYOKEN Y., MCKEEHAN W.L. & STEVENS J.I. (1991) Hepatocyte growth factor/hepatopoietin A stimulates the growth of rat kidney proximal tubular epithelial cells (RPTE), rat nonparenchymal liver cells, human melanoma cells, mouse keratinocytes and stimulates anchorage-independent growth of SV-40 transformed RPTE. Biochem. biophys. Res. Commun. 174, 331. 16. MATSUMOTO K., TAJIMA H. & NAKAMURA T. (1991) Hepatocyte growth factor is a potent stimulator of human melanocyte DNA synthesis and growth. Biochem. biophys. Res. Commun. 176,45.
149
17. MORIMOTO A., OKAMURA K., HAMANAKA R., SATO Y., SHIMA N., HIGASHIo K. & KUWANO M. (1991) Hepatocyte growth factor modulates migration and proliferation of human microvascular endothelial cells in culture. Biochem. biophys. Res. Commun. 179, 1042. 18. GOHDA E., TSUBOUCHI H., NAKAMURA H., HIRANO S., SAKIYAMA S., TAKAHASHI K., MIYAZAKI H., HASHIMOTO S. & DAKUHARA Y. (1988) Purification and partial characterization of hepatocyte growth factor from a patient with fulminant hepatic failure. J. clin. Invest. 81, 414. 19. SHIMIZU I., ICHIHARA A. & NAKAMURA T. (1 99 1) Hepatocyte growth factor in ascites from patients with cirrhosis. J. Biochem. 109, 14. 20. KINOSHITA T., TASHRO K. & NAKAMURA T. (1989) Marked increase of HGF mRNA in non-parenchymal liver cells of rats treated with hepatoxins. Biochem. biophys. Res. Commun. 165, 1229. 21. WEIDNER K.M., ARAKAKi N., HARTMANN G. VANDEKERCKHOVE J., WERDGART S., RIEDER H. et al. (1991) Evidence for the identity of human scatter factor and human hepatocyte growth factor. Proc. natl. Acad. Sci. U.S.A. 88, 7001. 22. Noji S., TASHIRo K., KOYAMA E., NOHNO T., OHYAMA K., TANIGUCHI S. & NAKAMURA T. (1990) Expression of hepatocyte growth factor gene in endothelial and Kupffer cells of damaged rat livers, as revealed by in situ hybridization. Biochem. biophys. Res. Commun. 173, 42. 23. HALLETT M.B., Luzio J.P. & CAMPBELL A.K. (1981) Stimulation of Ca2 + dependent chemiluminiscence in rat polymorphonuclear leukocytes by polystyrene beads and the non-lytic action of complement. Immunology, 44, 569. 24. AL-MOHANNA F.A. & HALLETT M.B. (1988) The use of fura-2 to determine the relationship between cytoplasmic free Ca2+ and oxidase activation in rat neutrophils. Cell Calcium, 9, 17. 25. ZARNEGAR R., MUGA S., RAHIJA R., MICHALOPOULOs G. (1990) Tissue distribution of hepatopoietin-A: a heparin-binding polypeptide growth factor for hepatocytes. Proc. natl. Acad. Sci. U.S.A. 87, 1252. 26. RUBIN J.S., CHAN A.M.L., BORTrARo D.P., BURGESS W.H., TAYLOR W.G., CECH A.C. et al. (1991) A broad-spectrum human lung fibroblast-derived mitogen is a variant of hepatocyte growth factor. Proc. natl. Acad. Sci. U.S.A. 88,414. 27. HUANG C-K., BONAK V., LARAMEE G.R. & CASNELLIE J.E. (1990) Protein tyrosine phosphorylation in rabbit peretoneal neutrophils. Biochem. J. 269,431. 28. PETERSEN M., STEADMAN R., HALLETT M.B., MATTHEWS N. & WILLIAMS J.D. (1990) Zymozan-induced leukotriene B4 generation by human neutrophils is augmented by rhTNF-a but not chemotactic peptide. Immunology, 70, 75. 29. RICHE J., ANDERSON T. & OLSSON L. (1988) Effect of tumor necrosis factor and granulocyte-macrophage colony-stimulating factor on neutrophil degranulation. J. Immunol. 142, 3199.
BRIEF COMMUNICATION
Neutrophil priming by hepatocyte growth factor,
a
novel cytokine
W. JIANG, M. C. A. PUNTIS, T. NAKAMURA* & M. B. HALLETT University Department of Surgery, University of Wales College of Medicine, Cardiff, U.K. and *Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan
Accepted for publication 12 April 1992
SUMMARY We demonstrate here that the recently defined cytokine hepatocyte growth factor (HGF) 'primes' human neutrophils. Recombinant human HGF over the concentration range 0-1-20 ng/ml increased the neutrophil response to f-met-leu-phe by up to 200%, and required only a short preincubation, 10 min producing the maximum effect. Priming was independent of changes in cytosolic-free calcium homeostasis. We conclude that HGF may be a physiologically important cytokine with 'priming' activity for neutrophils.
Activation of the non-mitochondrial oxidase of neutrophils, by generating toxic metabolites of oxygen, plays a crucial role in combating infection"2 but also causes tissue damage.3'4 This oxidative response may be enhanced by the activity of neutrophil 'primers', such as the cytokines tumour necrosis factor-ca (TNF-cc),5 interleukin-1 (IL-1),6 IL-6,7 IL-8,8 and granulocytemacrophage colony-stimulating factor (GM-CSF).9 Hepatocyte growth factor (HGF) is a recently defined growth factor which is a heterodimer molecule composed of a 69,000 MW a-subunit and a 34,000 MW f-subunit. This factor was first found in sera from partially hepatectomized rats,'0 its complementary DNA (cDNA) has recently been cloned from both the rat" and man.'2 HGF provokes DNA synthesis in hepatocytes in primary culture'3 and acts via a specific receptor on hepatocytes."4 HGF also affects keratinocytes, melanocytes and other cells.'5-'7 HGF has been reported to appear in the sera and other biological fluids in some diseases. 18-20 Recent reports have shown that HGF is identical to scatter factor which can dissociate epithelial cells.2' As HGF shares with other neutrophil primers both growth factor-like activity and a monocytic origin (from Kupffer cells),22 we have investigated whether HGF has priming activity for the neutrophil oxidase response. Human neutrophils were isolated from normal blood by dextran sedimentation and Ficoll-Hypaque centrifugation, and then resuspended in HEPES-buffered Krebs medium. The production of reactive oxygen metabolites was monitored using luminol-dependent chemiluminescence as reported previously.23 The cytosolic free Ca2+ was measured using dual wavelength fluorometry of Fura-2 (Spex CM, Instrument SA, Stanmore,
U.K.), as previously described.24 The cytosolic Fura-2 concentration of approximately 100 yM increased the cytosolic Ca2+ buffering capacity by 10-20%. Recombinant human hepatocyte growth factor was over 90% pure, half maximal activity on hepatocyte DNA synthesis was 3-75 ng/ml."I Stock solutions of 1 ug/ml were kept at -70° and diluted immediately before use. In order to determine the responsiveness of neutrophils to other 'primers', human recombinant IL-8 (89/520, NBSB, Potters Bar, U.K.) and TNF-a (a gift from Dr N. Matthews, UWCM, Cardiff, U.K.) were also used. Fura-2/AM was purchased from Molecular Probes (OR). All other materials were purchased from Sigma (Poole, Dorset, U.K.) unless otherwise stated. Priming occurred over a wide range of rHGF concentrations. Maximal priming was achieved by 10 ng/ml which enhanced the response by over 200%. The lowest detectable
240
C 190
0S t
140 ,
I
001
0-1
1
10
100
rHGF (ng/ml) Figure 1. The dose response of HGF priming on neutrophils. Neutrophils were preincubated for 10 min at 370 with rHGF. At the end of this period the luminol-dependent chemiluminescence rate was recorded before (0) and at the peak rate after stimulation (0) with FMLP (1 pM) plus cytochalasin B (5 pg/ml). The results are expressed as per cent priming, calculated as percentage of the response in the absence of rHGF.
Abbreviations: FMLP, f-met-leu-phe; GM-CSF, granulocytemacrophage colony-stimulating factor; HGF, hepatocyte growth factor; IFN-y, interferon-y; IL, interleukin; rHGF, recombinant hepatocyte growth factor; TNF-a, tumour necrosis factor-a. Correspondence: Dr W. Jiang, University Dept. of Surgery, University of Wales College of Medicine, Cardiff, U.K. :
147
148
W. Jiang et al. 800
800 r
Control
600
600
rHGF (50 ng/ml)
1 yilZ^q 1/ 5/il
Qb 200
0
Medium
FMLP
200
o*
rHFML
0 20 40 60 80 100 120 140 160 180 200 220 240 Time (second) Figure 2. Cytosolic-free Ca2 + changes in neutrophils. HGF was tested at concentrations 1, 10, 20, 30 and 50 ng/ml. No cytosolic-free calcium change was observed at these concentrations. At the first arrow either control medium or HGF (50 ng/ml) was added to the cell suspension. At the second arrow in each experiment FMLP (1 yM) was added.
0 20 40 60 80 100 120 140 160 180 200 220 240
enhancement occurred at 01 ng/ml (Fig. 1). In five separate experiments results show that the effect of preincubation with rHGF (5 ng/ml) for 10 min at 370, was to increase the production of reactive oxygen metabolites induced by the peptide f-met-leu-phe (FMLP, I tiM) plus cytochalasin B (5 jug/ ml) by 145 + 13-0% (P < 0-05 by Mann-Whitney U-test). This was comparable with IL-8 (2-0 ng/ml) and TNF-a (1 0 ng/ml) which prime neutrophils by 185-2+ 17-1 % and 136-4+ 19-4% respectively. Maximum priming by rHGF occurred rapidly, within 10 min, and remained at this level for up to 90 min (data not shown), suggesting that a protein synthesis step was not involved. If FMLP was absent no direct activation of the neutrophil oxidase was observed using concentrations of rHGF alone up to 20 ng/ml. It was unlikely that the effects reported here on neutrophils could be attributed to contaminating lipopolysaccharide (LPS) because heat (800, 30 min) abolished the priming effect (101 1-% after heat treatment of HGF) whereas polymixin B (1 mg/ml), which binds LPS, had no effects (149-0% in the presence of HGF and polymixin B). Furthermore, the priming effects were completely blocked by a specific rabbit anti-human HGF antibody (100-2% in the presence of HGF with anti-HGF). The effect on neutrophils was therefore attributed to rHGF. The possibility that priming by rHGF was mediated by a change in Ca2+ homoeostasis within the neutrophil was examined by measurement of the cytosolic-free Ca2+ concentration in neutrophils during incubation with HGF and during subsequent activation with FMLP. At concentrations up to 50 ng/ ml, HGF had no detectable effect on cytosolic-free Ca2 . Furthermore there was no detectable effect of rHGF on the peak, timing or kinetics of the transient cytosolic free Ca2+ rise induced by FMLP (Fig. 2). This eliminates the possibility that the mechanism of the activity of HGF was via up-regulation of FMLP receptors. The results of the experiments described here demonstrate a novel and previously undescribed property of HGF, namely the ability to prime the neutrophil oxidase response. HGF is a recently defined growth factor, synthesized by liver macrophages (Kupffer cells), endothelial cells and also expressed on
certain other tissues such as brain, lung, thyroid and pancreas.2225 It has major effects on hepatocytes, increasing DNA synthesis. We propose that HGF should, in addition, be added to the list of physiologically relevant neutrophil primers, which includes TNF-x, IL-1, IL-6, IL-8, GM-CSF, TGF-f3, IFN-y and substance P. It should be noted that although hepatocyte growth factor receptors have been detected on several cell and tissue types, such as hepatocytes, lung, kidney, spleen and most epithelial cells,'4 there are no reports at present which demonstrate their presence on neutrophils. Although the mechanisms by which HGF act are not yet established, Rubin et al.26 have recently reported that rHGF induces tyrosine phosphorylation in epithelial cells and melanocytes, and have suggested that the signalling pathway for this factor may involve the activation of tyrosine kinase. The possibility exists that HGF may act similarly in neutrophils and interact synergistically with FMLP-induced increased phosphotyrosine levels.27 HGF produces priming with a similar magnitude and a similar lack of effect on cytosolic-free Ca2+ homoeostasis as TNF-a, suggesting that these primers may share a common mechanism.2829 Further study will be required to establish this.
ACKNOWLEDGMENTS JWG thanks Scotia Pharmaceutical Ltd and MBH thanks the Arthritis and Rheumatism Council (U.K.) for support.
REFERENCES 1. FERRANTE A. (1989) Tumor necrosis factor alpha potentiates neutrophil antimicrobial activity: increased fungicidal activity against Torulopsis glabrata and Candida albicans and associated increases in oxygen radical production and lysosomal enzyme release. Infect. Immun. 57, 2115. 2. KUMARATILAKE L. M., FERRANTEA. & RZEPCZYK C. (199 1) The role of T lymphocytes in immunity to Plasmodiumfalciparum. Enhancement of neutrophil-mediated parasitic killing by lymphotoxin and IFN-gamma: comparisons with tumor necrosis factor effects. J. Immunol. 146, 762.
Neutrophil priming by hepatocyte growth factors 3. TELLADO J.M. & CHRISTOU V. (1991) Critically ill anergic patients demonstrate polymorphonuclear neutrophil activation in the intravascular compartment with decreased cell delivery to inflammatory focci. J. Leukoc. Biol. 50, 547. 4. KOWANKO I.C. & FERRANTE A. (1991) Granulocyte-macrophage colony-stimulating factor auguments neutrophil-mediated cartilage degradation and neutrophil adherence. Arthritis Rheum. 34, 1453. 5. KLEBANOFF S.J., VADAS M.M., HARLAN J.M., SPARKS L.H., GAMBLE J.R., AGoSTI J.M. & WALTERSDORPH A.M. (1986) Stimulation of neutrophils by tumor necrosis factor. J. Immunol. 136, 420. 6. FERRANTE A., NANDOSKAR M., WALZ A., GOH D.H.B. & KOWANKO I.C. (1988) Effects of tumour necrosis factor alpha and interleukin- l alpha and beta on human neutrophil migration, respiratory burst and degranulation. Int. Archs Allergy appl. Immunol. 86, 82. 7. BORISH L., ROSENBAUM R., ALBURY L. & CLARK S. (1989) Activation of neutrophils by recombinant interleukin-6. Cell. Immunol. 121, 280. 8. BAGGIOLINI M., WALZ A. & KUNKEL S.L. (1989) Neutrophilactivating peptide-1/interleukin-8, a novel cytokine that activates neutrophils. J. clin Invest 84, 1045. 9. MEGE J.L., GOMEZ-CAMBRONERO J., MOLSKi T.F.P., BECKER E.L. & SHA'AFI R.I. (1989) Effect of granulocyte-macrophage colonystimulating factor on superoxide production in cytoplasts and intact human neutrophils. J. Leukoc. Biol. 46, 161. 10. NAKAMURA T., NAWA K. & ICHIHARA A. (1984) Partial purification and characterization of hepatocyte growth factor from serum of hepatectomized rats. Biochem. biophys. Res. Commun. 122, 1450. 11. NAKAMURA T., NISHIZAWA T., HAGIYA M., SEKI T., SHIMONISHI M., SUGIMURA A., TASHIRO K. & SHIMIZU S. (1989) Molecular cloning and expression of human hepatocyte growth factor. Nature,
342,440. 12. TASHIRo K., HAGIYA M., NISHIZAWA T., SEKI T., SHIMONISHI M., SHIMIZU S. & NAKAMURA T. (1990) Deduced primary structure of rat hepatocyte growth factor and expression of the mRNA in rat tissues. Proc. natl. Acad. Sci. U.S.A. 87, 3200. 13. ZARNEGAR R. & MICHALOPOULOS G. (1989) Purification and biological characterization of human hepatopoietin A, a polypeptide growth factor for hepatocytes. Cancer Res. 49, 3314. 14. ZARNEGAR R., DEFRANCES M.C., OLIVE L. & MICHALOPOULOS G. (1990) Identification and partial characterization of receptor binding sites for HGF on rat hepatocytes. Biochem. biophys. Res. Commun. 173, 1179. 15. KAN M.K., ZHANG G., ZARNEGAR R., MICHALOPOULOS G., MYOKEN Y., MCKEEHAN W.L. & STEVENS J.I. (1991) Hepatocyte growth factor/hepatopoietin A stimulates the growth of rat kidney proximal tubular epithelial cells (RPTE), rat nonparenchymal liver cells, human melanoma cells, mouse keratinocytes and stimulates anchorage-independent growth of SV-40 transformed RPTE. Biochem. biophys. Res. Commun. 174, 331. 16. MATSUMOTO K., TAJIMA H. & NAKAMURA T. (1991) Hepatocyte growth factor is a potent stimulator of human melanocyte DNA synthesis and growth. Biochem. biophys. Res. Commun. 176,45.
149
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