Growth Factors, 1992, Vol. 6, pp. 179-186 Reprints availabledirectly from the publisher Photocopyingpermitted by license only

0 1992 Harwood Academic PublishersGmbH

Printed in the United Kingdom

Mini-Review

Granulocyte Colony-Stimulating Factor: Structure, Function and Physiology JUDITH E. LAYTON

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Melbourne Turnour Biology Branch, Ludwig Institute for Cancer Research, Melbourne, Australia, 3050

INTRODUCTION

was isolated by cross-hybridization with the human cDNA (Tsuchiya et al., 1986). Only one Granulocyte colony-stimulating factor (G-CSF) is mRNA species has been found in mice. The a member of a family of glycoproteins that are murine predicted protein was 178 amino acids required for the survival, proliferation and dif- and showed 72.8% sequence identity with human ferentiation of hemopoietic precursors and the G-CSF. activation and survival of mature hemopoietic Both the human and murine chromosomal cells (Metcalf, 1986; Clark and Kamen, 1987; genes for G-CSF have been isolated and Morstyn and Burgess, 1988). G-CSF was first sequenced (Nagata et al., 1986b; Tsuchiya et al., described as an activity that caused terminal dif- 1987). The two genes have very similar structures ferentiation of some myeloid leukemia cell lines with five exons and four introns. The exon (Burgess and Metcalf, 1980). It can also be dis- sequences are highly conserved but there is little tinguished from the other CSFs by its ability to homology between the intron sequences. The stimulate almost exclusively neutrophil colonies human gene has been mapped to the q21-22 from bone marrow precursors. Its actions on region of chromosome 17 (Kanda et al., 1987; mature hemopoietic cells appear to be restricted Simmers et al., 1987; Tweardy et al., 1987) and the murine gene is located on the distal half of to neutrophils. Murine G-CSF was first purified from con- the homologous murine chromosome 11 ditioned medium from the lungs of mice injected (Buckberg et al., 1988). with endotoxin (Nicola et al., 1983) and was found to be a glycoprotein of Mr 25,000. Two groups independently purified human G-CSF G-CSF PROTEIN STRUCTURE from carcinoma cell lines that constitutively secrete G-CSF Welte et al. (1985) used a bladder The 174 amino acid form of human G-CSF (Fig. 1) carcinoma line (5637) and Nomura et al. (1986) is the most abundant form in 5637 and CHU-2 used a squamous carcinoma cell line (CHU-2). cells (Nagata et al., 1986; Zsebo et al., 1986). It is The purified proteins were of Mr 18,000 and not known whether the 177 amino acid form is 19,000, respectively. N-terminal sequences restricted to carcinoma cells or is also found in obtained from the purified proteins were used to normal cells. Both human and murine G-CSF design oligonucleotide probes for cloning human contain five cysteine residues which would be G-CSF cDNAs (Nagata et al., 1986a; Souza et al., expected to form two disulphide bonds and one 1986). Two different cDNA clones were ident- free cysteine. Stqdies of human G-CSF by Lu et ified which coded for mature proteins of 174 and al. (1989) established that the Cys at position 17 177 amino acid residues (Nagata et al., 1986b; was free and was inaccessible in t-he native molZsebo et al., 1986). The two mRNAs were gener- ecule. They found two disulphide bridges ated by alternative splicing of a single precursor assigned as Cys36-Cys42 and Cys64-Cys74. Four RNA (Nagata et al., 1986b). Murine G-CSF cDNA of the cysteines in murine G-CSF are conserved 179

LAYTON

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and form disulphide bonds equivalent to the human whereas the fifth free cysteine is at position 86. The apparent molecular weight of G-CSF purified from carcinoma cell supernatants was higher 'than that of recombinant G-CSF produced in E . coZi (19,600 compared with 18,800; Souza et al., 1986). A likely explanation for the difference was that the G-CSF produced in mammalian cells was glycosylated. Since G-CSF does not contain asparagine, there are no potential N-glycosylation sites thus the molecule is probably O-glycosylated. Souza et al. (1986) treated G-CSF from 5637 cells with neuraminidase and 0-glycanase and found that its apparent molecular weight was reduced to 18,000, confirming the O-glycosylation. Kubota et al. (1990) established that the glycosylation site was Thr133 in human G-CSF. Although glycosylation did not affect the specific activity of G-CSF in vitro, it enhanced the stability of the molecule and prevented loss of activity through aggregation (Oheda et al., 1990). It is not yet clear whether the enhanced stability of glycosylated G-CSF affects its activity in v i m compared with non-glycosylated G-CSF. The tertiary structure of G-CSF has not yet been determined although the production of

crystals of a mutant G-CSF suitable for X-ray diffraction studies has been reported (Nagahara et al., 1990). Circular dichroic spectroscopy results indicated that G-CSF contains a high proportion (69%) of a-helical structure (Lu et al., 1989). Algorithms predicting a-helical regions and amphipathic helix searches have been used to predict the secondary structure of G-CSF (Parry et al., 1988, 1991; Bazan, 1990). These two predictions are quite similar (Fig. 21, the main differences being in the length of the first a-helix and an additional region predicted by Parry et al. (1988). G-CSF appears to belong to a group of cytokines including the other CSFs, interleukins 2-7, erythropoietin, growth hormone and prolactin predicted to have a 4-a-helical bundle structure (Bazan, 1990; Parry et al., 1991). The arrangement of the 4 helices in the folded molecule is unknown except for growth hormone (iIlustrated in Bazan, 1990). A slightly different arrangement has been proposed by Parry et al. (1991). STRUCTURE-FUNCTION RELATIONSHIPS

Two approaches have been used in order to identify regions of G-CSF that have functional 0

1

40

120

110

100

FIGURE 1. The amino acid sequence of human G-CSF showing the position of the disulphide bonds (Luet al., 1989).

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G-CSF S'IXUCTURE/FUNCTION

importance. Kuga et al. (1989) made a series of mutants using a variety of techniques. They found that the N-terminal 11 residues were not essential for biological activity. However, substitutions in four of these residues together with Cysl7 resulted in a mutein (KW-2228) with enhanced biological activity. The mutein KW2228 had enhanced thermal stability, possibly increased protease resistance (Okabe et al., 1990) and apparently a lower receptor dissociation constant (Uzumaki et al., 1988). If this mplecule is confirmed to have lower dissociation constant, it would suggest that the N-terminus can modulate the conformation of the G-CSF receptor binding determinants. Other alterations in the molecule between residue 18 and the C-terminus all resulted in loss of biological activity. Many of these mutants were deletions or tandem repeats which would 'be expected to substantially alter the tertiary structure of the molecule and thus lead indirectly to loss of biological activity. Deletion or substitution of Leu35 also caused loss of biological activity. These single residue alterations are less likely to affect the tertiary structure and therefore suggest that this region is involved in receptor binding. Since the mutants were only partially purified and were not structurally characterized, the conclusions from this study require confirmation. There is also evidence that the region at Leu35 is important from the observations of Nagata et al. (1986b). They found that the 177 residue form of G-CSF, which contains an additional three amino acids between Leu35 and Cys36, had less activity than the 174

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residue form although the difference was not accurately quantitated. We used an immunochemical approach to define functionally important regions of G-CSF. We produced monoclonal antibodies to G-CSF and mapped the binding sites of distinct groups of antibodies that recognized different epitopes on G-CSF (Layton et al., 1991, in press). The binding of seven out of eight neutralizing antibodies was dependent on the conformation of G-CSF, therefore chemical and enzymatic digestion of unreduced G-CSF as well as peptide synthesis were used to generate peptides to test for antibody binding. The binding of all the neutralizing antibodies was localized to residues 20-46, which strongly supports the previous observations that the region around Leu35 might be important for receptor binding. The other non-neutralizing antibodies recognized regions throughout the molecule, including the N-terminus, indicating that these regions are not directly involved in receptor binding. None of the antibodies appeared to bind close to the C-terminus, so the involvement of the C-terminus in receptor binding is unknown. The C-termini of several structurally related cytokines have been implicated in receptor binding. In the case of IL-6, which is the cytokine most closely related to G-CSF, both deletion studies (Brakenhoff et al., 1990; Kruttgen et al., 1990) and the mapping of neutralizing antibody binding sites (Ida et al., 1989; Brakenhoff et al., 1990) indicated the importance of the C-terminus for function. Cunningham et al. (1989) found that

Bazan 1990 10

35

70

93 102

69

93103

124

145

173

Parry et al. 1988 13 24

NH2

46 54

127

152

172

COOH

FIGURE 2. Predicted a-helical regions of G-CSF (shaded areas). A comparison of the predictions of Bazan (1990) and Parry et al. (1988). The disulphide bonds are indicated by lines below the bars.

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LAYTON

the C-terminus is part of the receptor binding site of human growth hormone and studies of Fibi et al. (1991) implicated the C-terminus of erythropoietin. Studies of GM-CSF (Clark-Lewis et al., 1988; Nice et al., 1990; Kanakura et al., 1991) and IL-3 (Ziltener et al., 1987; Lokker et al., 1991) have concluded that the C-terminus is one of several regions important for receptor binding of these cytokines. A recent report from Le et al. (1991) has indicated that the C-terminus of human IL-4 is important for its function. There is also some evidence to indicate that the C-terminus of leukemia inhibitory factor (LIF) is required for function. A clone of murine LIF that was missing a region encoding the nine C-terminal residues was inactive when expressed in yeast (Gearing et al., 1987). Thus, the involvement of the C-terminus in function appears to be a common feature of cytokines and it seems likely that the C-terminus of G-CSF will be found to be important.

(Roilides et al., 1991). Studies of Wang et al. (1988) demonstrated the chemotactic activity of G-CSF for human granulocytes and monocytes and G-CSF has been reported to induce neutrophi1 adhesion (Okada et al., 1990; Yuo et al., 1990). G-CSF may therefore affect the migration of neutrophils into the tissues. Presumably, these in vitro functions reflect the in vivo role of G-CSF in maintenance of steady state hemopoiesis and defence against infection.

In vivo Activities of G-CSF In contrast to the in vitro situation, the normal in vivo role of G-CSF is less well defined. CSFs are

likely to be involved in steady state production of hemopoietic cells in the bone marrow and in the responses to infection and other inflammatory conditions. The CSFs and other cytokines that have activity in vitro on hemopoietic progenitors (interleukins 1-7, c-kit ligand) to some extent have overlapping activities but may be produced in different circumstances. G-CSF is produced by several different cell PHYSIOLOGY OF G-CSF types in vitro. Bone marrow stromal cell cultures have been reported to secrete low levels of G-CSF In vitro Activities in the absence of stimulation (Migliaccio et al., The actions of G-CSF in vitro on mature neutro- 1990), but this production appears to be transient philic granulocytes and their precursors have and a result of endogenously produced IL-1 been described in detail (recently reviewed by (Fibbe et al., 1988a). Stromal cells have been Nicola, 1990). Briefly, both human and murine G- shown to secrete G-CSF in response to IL-1 (Fibbe CSF stimulate the growth of neutrophil colonies et al., 1988a; Yang et al., 1988) or after transformfrom bone marrow precursors although at high ation (Rennick et al., 1987; Nemunaitis et al., concentrations, some macrophages are also 1989; Novotny et al., 1990). Bone marrow derived found (Metcalf and Nicola, 1983; Welte et al., macrophages and peripheral blood monocytes 1985; Souza et al., 1986; Nomura et al., 1986; secrete G-CSF in response to IL-1 or LPS (Fibbe et Zsebo et al., 1986). G-CSF is able to increase the al., 1986; Vellenga et al., 1988). Similarly, endosurvival of murine precursors of other lineages thelial cells (Broudy et al., 1987; Seelentag et al., without apparently causing proliferation 1987; Zsebo et al., 1988; Segal and Bagby, 1988) (Metcalf and Nicola, 1983). The more mature and fibroblasts (Koeffler et al., 1987; Fibbe et al., human neutrophilic promyelocytes were also 1988b; Kaushansky et al., 1988; Leizer et al., 1990) shown to proliferate to a limited extent in stimulated with IL-1 or TNFa secrete G-CSF. response to G-CSF (Begley et al., 1988). Other cell types reported to secrete G-CSF after G-CSF enhances the survival of mature human stimulation are articular cartilage and chondroneutrophils at low concentrations (Begley et al., cytes (Campbell et al., 1991) and astroglial cells 1986) and activates or primes mature cell func- (Tweardy et al., 1990; Tweardy et al., 1991). tions such as the respiratory burst in human neu- Because all these cell types require stimulation trophils (Kitagawa et al., 1987; Sullivan et al., by IL-I, TNFa or LPS rather than exhibiting 1987; Nathan, 1989), antibody-dependent cell- constitutive secretion, their production of G-CSF mediated cytotoxicity in both murine (Lopez et may be more important in the response to an al., 1983) and human (Vadas et al., 1983) neutro- environmental insult such as infection rather phils and phagocytosis by human neutrophils than in maintaining normal neutrophil numbers.

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G-CSF S T R U W C T I O N

On the other hand, experiments in dogs provide good evidence that G-CSF is required for maintenance of normal circulating neutrophil levels. Hammond et al. (1991) found that dogs being administered human G-CSF became neutropenic. Investigation of this effect revealed that the dogs made antibodies to the human G-CSF which cross-reacted with, and neutralized, canine G-CSF. When the antibody levels declined, the neutrophil levels returned to normal, thus indicating that G-CSF was required for neutrophi1 production in normal animals. G-CSF is normally undetectable or found at only very low levels in the serum of healthy individuals (Watari et al., 1989; Shirafuji et al., 1989; Kawakami et al., 1990; Cebon et al., submitted). Local production in the bone marrow therefore would seem to be sufficient for normal maintenance of neutrophil levels. Serum G-CSF levels are elevated in response to infection. Early studies detected increased colony stimulating activity that probably included G-CSF in sera of patients (Foster et al., 1968; Metcalf, 1981) and mice (Trudgett et al., 1973) with infections. More recently, production of G-CSF in response to infection has been detected with more specific assays (Kawakami et al., 1990; Cheers et al., 1988; Cebon et al., submitted). Lipopolysaccharide (LPS),a product of Gram negative bacteria, has been shown to be a potent stimulator of G-CSF production (Metcalf, 1982; Golde and Cline, 1975; Quesenberry et al., 1972). LPS also stimulates secretion of IL-1 and TNFa which in turn activate a variety of cell types (endothelial cells, fibroblasts and monocytes) to secrete G-CSF. However, increased serum G-CSF is not restricted to Gram negative infections (Kawakami et al., 1990) and thus other bacterial products (Robinson et al., 1977) and secondary mediators are likey to stimulate G-CSF production during infection. When recombinant G-CSF was given directly to normal animals and cancer patients, a rapid (within 24 hr), dose-dependent elevation of circulating neutrophils was found (reviewed by Morstyn et al., 1989). Elevated neutrophil levels returned to normal within 24-48 hr of ceasing G-CSF administration, consistent with a direct effect of G-CSF on neutrophil production. The kinetics of human granulopoiesis after G-CSF treatment have been studied by using radioactive labelling techniques (Lord et al., 1989). The results indicated ihat the marrow transit time for

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maturing granulocytes was substantially reduced by G-CSF, since labelled cells appeared in the circulation in one day compared with the normal four days. In addition, an increase of 3.2 extra amplification divisions was calculated to occur during neutrophil development. The half life of circulating neutrophils was not affected. Thus, a relatively modest increase in the number of cell divisions can lead to approximately a ten-fold increase in circulating neutrophils. Similar results were found in mice (Lord et al., 1991). The factors that negatively regulate neutrophil production are not well understood. When an infection is controlled, serum G-CSF levels return to normal (Kawakami et al., 19901, probably because of the removal of the bacterial products that directly or indirectly stimulated G-CSF production. Other factors may also down-regulate G-CSF levels. In clinical trials in which patients were given continuous infusions of G-CSF, serum G-CSF was found to decrease to undetectable levels after the first three days of the infusion. The decrease correlated with the increase in neutrophils and did not occur in patients who were neutropenic after high dose chemotherapy and bone marrow transplantation, suggesting that the neutrophils were responsible for accelerated clearance of G-CSF (Layton et al., 1989). It is not known whether the increased clearance was directly mediated by neutrophils, for example through receptor binding, or whether an indirect effect was involved. CONCLUSIONS There is now good evidence that G-CSF is required for normal neutrophil production in vim. It is produced at elevated levels in response to infection, giving rise to increased neutrophil numbers and contributing to their activation, thus helping to control the infection. It is likely that future experiments such as G-CSF gene disruption and production of animals lacking G-CSF will add to our understanding of the importance of G-CSF in vivo. Mechanisms of down-regulation of neutrophil production need to be more fully explored. When the crystal structure of G-CSF is determined, it will provide a firmer basis for further experiments aimed at defining; the receptor binding site and the nature of the Ynteraction with the

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receptor. Such information should enable the development of agonists for improved clinical use and antagonists, which may be useful in controlling some leukemias and inflammatory diseases.

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REFERENCES Bazan, J. F. (1990) Haemopoietic receptors and helical cytokines. Immunol. Today 11,350-355. Begley, C. G., Lopez, A. F., Nicola, N. A., Warren, D. J., Vadas, M. A., Sanderson, C. J. and Metcalf, D. (1986) Purified colony-stimulating factors enhance the survival of human neutrophils and eosinophils in vitro: A rapid and sensitive microassay for colony-stimulating factors. Blood 68,162-166. Begley, C. G., Nicola, N. A. and Metcalf, D. (1988) Proliferation of normal human promyelocytes and myelocytes after a single pulse stimulation by purified GM-CSF or G-CSF. Blood 71,640-645. Brakenhoff, J. P. J., Hart, M., De Groot, E. R., Di Padova, F. and Aarden, L. A. (1990) Structure-function analysis of human IL-6. Epitope mapping of neutralizing monoclonal antibodies with amino- and carboyxl-terminal deletion mutants. 1.lmmunol. 145,561-568. Broudy, V. C., Kaushansky, K., Harlan, J. M. and Adamson, J. W. (1987) Interleukin 1 stimulates human endothelial cells to produce granulocyte-macrophage colony-stimulating factor and granulocyte colony-stimulating factor. 1. lrnrnunol. 139,464-468. Buckberg, A. M., Bedigan, H. G., Taylor, B. A., Brownell, E., Ihle, J. N., Nagata, S., Jenkins, N. A. and Copeland, N. G. (1988) Localization of Evi-2 to chromosome 11: linkage to other proto-oncogene and growth factor loci using interspecific backcross mice. Oncogene Res. 2,149-165. Burgess, A. W. and Metcalf, D. (1980) Characterization of a serum factor stimulating the differentiation of myelomonocytic leukemic cells. Int. 1. Cancer 39,647-654. Campbell, 1. K., Novak, U., Cebon, J., Layton, J. E. and Hamilton, J. A. (1991) Human articular cartilage and chondrocytes produce hemopoietic colony-stimulating factors in culture in response to IL-1.1. Immunol. 147,1238-1246. Cheers, C., Haigh, A. M., Kelso, A., Metcalf, D., Stanley, E. R. and Young, A. M. (1988) Production of colony-stimulating factors (CSFs) during infection: Separate determinations of macrophage-, granulocyte-, granulocyte-macrophage- and multi-CSFs. Infect. Immun. 56,247-251. Clark, S. C. and Kamen, R. (1987) The human hematopoietic colony-stimulating factors. Science 236,1229-1237. Clark-Lewis, I., Lopez, A. F., To, L. B., Vadas, M. A., Schrader, J. W., Hood, L. E. and Kent, S. B. H. (1988) Structure-function studies of human granulocyte-macrophage colonystimulating factor. Identification of residues required for activity. J. lmmunol. 141,881-889. Cunningham, B. C., Jhurani, P., Ng, P. and Wells, J. A. (1989) Receptor and antibody epitopes in human growth hormone identified by homolog-scanning mutagenesis. Scimce 243, 1330-1 336. Fibbe, W. E., van Damme, J., Billiau, A., Voogt, P. J., Duinkerken, N., Kluck, P. M. C. and Falkenburg, J. H. F. (1986) Interleukin-1 (22-K factor) induces release of granulocyte-macrophage colony-stimulating activity from human mononuclear phagocytes. Blood 68,1316-1321. Fibbe, W. E., van Damme, J., Billiau, A., Goselink H. M., Voogt, P. J., van Eeden, G., Ralph, P., Altrock, B. W. and

Falkenburg, J. H. F. (1988a) Interleukin 1 induces human marrow stromal cells in long-term culture to produce granulocyte colony-stimulating factor and macrophage colonystimulating factor. Blood 71,430-435. Fibbe, W. E., van Damme, J., Billiau, A., Duinkerken, N., Lurvink, E., Ralph, P., Altrock, B. W., Kaushansky, K., Willemze, R. and Falkenburg, J. H. F. (1988b) Human fibroblasts produce granulocyte-CSF, macrophage-CSF, and granulocyte-macrophage-CSF following stimulation by interleukin-1 and poly(rI).poly(rC). Blood 72,860-866. Fibi, M. R., Stiiber, W., Hintz-Obertreis, P., Liiben, G., Krumwieh, D., Siebold, B., Zettlmeissl, G. and Kiipper, H. A. (1991) Evidence for the location of the receptor-binding site of human erythropoietin at the carboxy-terminal domain. Blood 77,1203-1210. Foster Jnr., R., Metcalf, D., Robinson, W. A. and Bradley, T. R. (1968) Bone marrow colony stimulating activity in human sera. Results of two independent surveys in Buffalo and Melbourne. Brit. 1.Haematol. 15,147-159. Gearing, D. P., Gough, N. M., King, J. A., Hilton, D. J., Nicola, N. A., Simpson, R. J., Nice, E. C., Kelso, A. and Metcalf, D. (1987) Molecular cloning and expressing of cDNA encoding a murine myeloid leukemia inhibitory factory (LIF). EMBO 1.6,399!j-4002. Golde, D. W. and Cline, M. J. (1975) Endotoxin-induced release of colony-stimulating activity in man. Proc. SOC. E x p . Biol. Med. 149,845-848. Hammond, W. P., Csiba, E., Canin, A., Hockman, H., Souza, L. M., Layton, J. E. and Dale, D. C. (1991) Chronic neutropenia. A new canine model induced by human granulocyte colony-stimulating factor. 1.Clin. Invest. 87,704-710. Ida, N., Sakurai, S., Hosaka, T., Hosoi, K., Kunitomo, T., Shimazu, T., Maruyama, T., Matsuura, Y. and Kohase, M. (1989) Establishment of strongly neutralizing monoclonal antibody to human interleukin-6 and its epitope analysis. Biochem. Biophys. Res. Cornmun. 165,728-734. Kanakura, Y., Cannistra, S. A., Brown, C. B., Nakamura, M., Seelig, G. F., Prosise, W. W., Hawkins, J. C., Kaushansky, K. and Griffin, J. D. (1991) Identification of functionally distinct domains of human granulocyte-macrophage colonystimulating factor using monoclonal antibodies. Blood 77, 1033-1043. Kanda, N., Fukushige, S-I., Murotsu, T., Yoshida, M. C., Tsuchiya, M., Asano, S., Kazior, Y. and Nagata, S. (1987) Human gene coding for granulocyte-colony stimulating factor is assigned to the q21-q22 region of chromosome 17. Somat. Cell. Mol. Genet. 13,679-684. Kaushansky, K., Lin, N. and Adamson, J. W. (1988) Interleukin 1 stimulates fibroblasts to synthesize granulocytemacrophage and granulocyte colony-stimulating factors. Mechanism for the hematopoietic response to inflammation. 1.Clin. Invest. 81,92-97. Kawakami, M., Tsutsumi, H., Kumakawa, T., Abe, H., Hirai, M., Kurosawa, S., Mori, M. and Fukushima, M. (1990) Levels of serum granulocyte colony-stimulating factors in patients with infections. Blood 76,1962-1964. Kitagawa, S., Yuo, A,, Souza, L. M., Saito, M., Miura, Y. and Takaku, F. (1987) Recombinant human granulocyte colonystimulating factor enhances superoxide release in human granulocytes stimulated by the chemotactic peptide. Biochem. Biophys. Res. Cornmun. 144,1143-1146. Koeffler, H. P., Gasson, J., Ranyard, J., Souza, L., Shepard, M. and Munker, R. (1987) Recombinant human TNFa stimulates production of granulocyte colony-stimulating factor. Blood 70’55-59. Kriittgen, A., Rose-John, S., Dufhues, G., Bender, S., Liitticken, C., Freyer, P. and Heinrich, P. C. (1990) The three carboxy-terminal amino acids of human interleukin-6 are essential for its biological activity. F E B S Letters 273,95-98. Kubota, N., Orita, T., Hattori, K., Oh-eda, M., Ochi, N. and

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G-CSF STRUCTURE/FIR\ICTION

Yamazaki, T. (1990) Structural characterization of natural and recombinant human granulocyte colony stimulating factors. J. Biochem (Tokyo) 107,486-492. Kuga, T., Komatsu, Y., Yamasaki, M., Sekine, S., Miyaji, H., Nishi, T., Sato, M., Yokoo, Y., Asano, M., Okabe, M., Morimoto, M. and Itoh, S. (1989) Mutagenesis of human granulocyte colony stimulating factor. Biochem. Biophys. Res. Commun. 159,103-111. Layton, J. E., Hockman, H., Sheridan, W. P. and Morstyn, G. (1989) Evidence for a novel in viuo control mechanism of granulopoiesis: Mature cell-related control of a regulatory growth factor. Blood 74,1303-1307. Layton, J. E., Morstyn, G., Fabri, L. J., Reid, G. E., Burgess, A. W., Simpson, R. J. and Nice, E. C. Identification of a functional domain of human granulocyte colony-stimulating factor using neutralizing monoclonal antibodies. J. Biol. Chem. In press. Le, H. V., Seelig, G. F., Syto, R., Ramanathan, L., Windsor, W. T., Borkowski, D. and Trotta, P. P. (1991) Selective proteolytic cleavage of recombinant human interleukin 4. Evidence for a critical role of the C-terminus. Biochemistry 30, 9576-9582. Leizer, T., Cebon, J., Layton, J. E. and Hamilton, J. A. (1990) Cytokine regulation of colony-stimulating factor production in cultured human synovial fibroblasts: I. Induction of GM-CSF and G-CSF production by interleukin-1 and tumor necrosis factor. Blood 76,1989-1996. Lokker, N. A., Zenke, G., Strittmatter, U., Fagg, B. and Mowa, N. R. (1991) Structure activity relationship study of human interleukin 3: role of the C-terminal region for biological activity. EMBO J. 10,2125-2131. Lopez, A. F., Nicola, N. A., Burgess, A. W., Metcalf, D., Battye, F. L., Sewell, W. A. and Vadas, M. (1983) Activation of granulocyte cytotoxic function by purified mouse colony-stimulating factors. J.Immunol. 131,2983-2988. Lord, B. I., Bronchud, M. H., Owens, S., Chang, J., Howell, A., Souza, L. and Dexter, T. M. (1989) The kinetics of human granulopoiesis following treatment with granulocyte colony-stimulating factor in uivo. Proc. Natl. Acad. Sci. U S A 86, 9499-9503. Lord, 8. I., Molineux, G., Pojda, Z., Souza, L. M., Mermod, J.-J. and Dexter, T. M. (1991) Myeloid cell kinetics in mice treated with recombinant interleukin-3, granulocyte colony-stimulating factor (CSF), or granulocyte-macrophage CSF in vivo. Blood 77,2154-2159. Lu, H. S., Boone, T. C., Souza, L. M. and Lai, P-H. (1989) Disulfide and secondary structures of recombinant human granulocyte colony stimulating factor. Arch. Biochem. Biophys. 268,Bl-92. Metcalf, D. (1981) Induction of differentiation in murine myelomonocytic leukemia cells by the serum of patients with acute myeloid leukemia and other diseases. Int. J. CanC ~ T27,577-584. Metcalf, D. (1982) Sources and biology of regulatory factors active on mouse myeloid leukemic cells. J. Cell. Physiol. S U P P ~1,175-183. . Metcalf, D. (1986) The molecular biology and function of the granulocyte-macrophage colony-stimulating factors: Review. Blood 67,257-267. Metcalf, D. and Nicola, N. A. (1983) Proliferative effects of purified granulocyte colony-stimulating factor (G-CSF) on normal mouse hemopoietic cells. J. Cell. Physiol. 116, 198-206. Migliaccio, A. R., Migliaccio, G., Johnson, G., Adamson, J. W. and Torok-Storb, B. (1990) Comparative analysis of hematopoietic growth factors released by stromal cells from normal donors or transplanted patients. Blood 75,305-312. Morstyn, G. and Burgess, A. W. (1988) Hemopoietic growth factors: A review. Cancer Res 48,5624-5637. Morstyn, G., Lieschke, G. J., Sheridan, W., Layton, J. and

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Cebon, J. (1989) Pharmacology of the colony-stimulating factors. TiPS 10,154-159. Nagahara, Y., Konishi, N., Yokoo, Y. and Hirayama, N. (1990) Crystallization and preliminary diffraction studied of recombinant human granulocyte-stimulating factor (KW2228).J. Mol. Biol. 214,25-26. Nagata, S., Tsuchiya, M., Asano, S., Kaziro, Y., Yamazaki, T., Yamamoto, O., Hirata, Y., Kubota, N., Oheda, M., Nomura, H. and Ono, M. (1986a) Molecular cloning and expression of cDNA for human granulocyte colony-stimulating factor. Nature 319,415418. Nagata, S., Tsuchiya, M., Asano, S., Yamamoto, O., Hirata, Y., Kubota, N., Oheda, M., Nomura, H. and Yamazaki, T. (1986b) The chromosomal gene structure and two mRNAs for human granulocyte colony-stimulating factor. EMBO J. 5,575-581. Nathan, C. F. (1989) Respiratory burst in adherent human neutrophils: Triggering by colony-stimulating factors CSFGM and CSF-G. Blood 73,301-306. Nemunaitis, J., Andrews, D. F., Crittenden, C., Kaushansky, K. and Singer, J. W. (1989) Response of simian virus 40 (SV40)-transformed, cultured human marrow stromal cells to hematopoietic growth factors. 1. Clin. Invest. 83,593-601. Nice, E., Dempsey, P., Layton, J., Morstyn, G., Cui, D-F., Simpson, R., Fabri, L. and Burgess, A. W. (1990) Human granulocyte-macrophage colony-stimulating factor (hGMCSF): Identification of a binding site for a neutralizing antibody. Growth Factors 3,159-169. Nicola, N. (1990) Granulocyte colony-stimulating factor. In Colony Stimulating Factors: Molecular and Cellular Biology, T. M. Dexter, J. M. Garland and N. G. Testa, eds, Marcel Decker, Inc., New York and Basel, pp. 77-109. Nicola, N. A., Metcalf, D., Matsumoto, M. and Johnson, G. R. (1983) Purification of a factor inducing differentiation in murine myelomonocytic leukemia cells. Identification as granulocyte colony-stimulating factor. J. Biol. Chem. 258, 9017-9023. Nomura, H., Imazeki, I., Oheda, M., Kubota, N., Tamura, M., Ono, M., Ueyama, Y. and Asano, S. (1986) Purification and characterization of human granulocyte colony stimulating EMBO J.5,871-876. factor (G-CSF). Novotny, J. R., Duehrsen, U., Welch, K., Layton, J. E., Cebon, J. S. and Boyd, A. W. (1990) Cloned stromal cell lines d m v e d from human Whitlock/Witte-type long-term bone marrow cultures. Exp. Hematol. 18,775-784. Oheda, M., Hasegawa, M., Hattori, K., Kuboniwa, H., Kojima, T., Orita, T., Tomonou, K., Yamazaki, T. and Ochi, N. (1990) 0-linked sugar chain of human granulocyte colony-stimulating factor protects it against polymerization and denaturation allowing it to retain its biological activity. J. Biol. C h m . 265,11432-11435. Okabe,M., Asano, M., Kuga, T., Komatsu, Y., Yamasaki, M., Yokoo, Y., Itoh, S., Morimoto, M. and Oka, T. (1990) In vitro and in vivo hematopoietic effect of mutant and human granulocyte colony-stimulating factor. Blwd 75,1788-1793. Okada, Y., Kawagishi, M. and Kusaka, M. (1990) Effect of recombinant human granulocyte colony-stimulating factor on human neutrophil adherence in vitro. Experientia 46, 1050-1053. Parry, D. A. D., Minasian, E. and Leach, S. J. (1988) Conformational homologies among cytokines: Interleukins and colony stimulating factors. J. Mol. Recog. 1,107-110. Parry, D. A. D., Minasian, E. and Leach, S. J. (1991) Cytokine conformations: Predictive studies. 1. Mol. Recog. In press. Quesenberry, P. J., Morley, A., Stohlman, F., Rickard, K., Howard, D. and Smith, M. (1972) Effect of endotoxin on granulopoiesis and colony-stimulating factor. N. Engl. J. Med. 286,227-232. Rennick, D., Yang, G., Gemmell, L. and Lee, F. (1987) Control of hemopoiesis by a bone marrow stromal cell clone: Lipo-

Growth Factors Downloaded from informahealthcare.com by University of Auckland on 12/05/14 For personal use only.

186

LAYTON

polysaccharide- and interleukin-1-inducible production of colony stimulating factors. Blood 69,682-691. Robinson, W. A., Entringer, M. A., Bolin, R. W. and Sbnington Jr., 0. G. (1977) Bacterial stimulation and granulocyte inhibition of granulopoietic factor production. N . Engl. J. Med. 297,1129-1134. Roilides, E., Walsh, T. J., Pizzo, P. A. and Rubin, M. (1991) Granulocyte colony-stimulating factor enhances the phagocytic and bactericidal activity of normal and defective human neutrophils. J. lnf. Diseuses 163,579-583. Seelentag, W. K., Mermod, J-J., Montesano, R. and Vassalli, P. (1987) Additive effects of interleukin-1 and tumor necrosis factor-a on the accumulation of the three granulocyte and macrophage colony-stimulating factor mRNAs in human endothelial cells. EMBO J. 6,2261-2265. Segal, G. M. and Bagby, Jr., G. C. (1988) Vascular endothelial cells and hematopoietic regulation. Int. 1. Cell Clon. 6, 306-312. Shirafuji, N., Asano, S., Matsuda, S., Watari, K., Takaku, F. and Nagata, S. (1989) A new bioassay for human granulocyte colony-stimulating factor (hGCSF) using murine myeloblastic NFS-60 cells as targets and estimation of its levels in sera from normal healthy persons and patients with infectious and hematological disorders. Exp. Hematol. 17,116-119. Simmers, R. N., Webber, L. M., Shannon, M. F., Garson, 0.M., Wong, G., Vadas, M. A. and Sutherland, G. R. (1987) Localization of the G-CSF gene on chromosome 17 proximal to the breakpoint in the t (15; 17) in acute promyelocytic leukemia. Blood 70,330-332. Souza, L. M., Boone, T. C., Gabrilove, J., Lai, P. H., Zsebo, K. M., Murdock, D. C., Chazin, V. R., Bruszewski, J., Lu, H., Chen, K. K., Barendt, J., Platzer, E., Moore, M. A. S., Mertelsman, R. and Welte, K. (1986) Recombinant human granulocyte colony-stimulating factor: Effects on normal and leukemic myeloid cells. Science 232,61-65. Sullivan, R., Griffin, J. D., Simons, E. R., Schafer, A. I., Meshulam, T., Fredette, J. P., Maas, A. K., Gadenne, A-S., Leavitt, J. L. and Melnick, D. A. (1987) Effects of recombinant human granulocyte and macrophage colony-stimulating factors on signal transduction pathways in human granulocytes. J. Immunol. 139,3422-3430. Trudgett,, A., McNeill, T. A. and Killen, M. (1973) Granulocyte macrophage precursor cell and colony-stimulating factor response of mice infected with Salmont+a typhimurium. Infect. Immun. 8,450-455. Tsuchiya, M., Asano, S., Kaziro, Y. and Nagata, S. (1986) Isolation and characterization of the cDNA for murine granulocyte colony-stimulating factor. Proc. Nutl. Acnd. Sci. USA 83,7633-7637. Tsuchiya, M., Kaziro, Y. and Nagata, S. (1987) The chromosomal gene structure for murine granulocyte colony-stimulating factor. Eur. 1.Biochem. 165,7-12. Tweardy, D. J., Cannizzaro, L. A., Paiumbo, A. P., Shane, S., Heubner, K.,Vantuinen, P., Ledbetter, D. H., Finan, J. B., Nowell, P. C. and Rovera, G. (1987) Molecular cloning and characterization of a cDNA for human granulocyte colonystimulating factor (G-CSF) from a glioblastoma multiforme cell line and localization of the G-CSF gene to chromosome band 17q21. Oncogene Res. 1,209-220. Tweardy, D. J., Mott, P. L. and Glazer, E. W. (1990) Monokine

modulation of human astroglial cell production of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. I. Effects of &la and ILIs.I. lmmunol. 144,2233-2241. Tweardy, D. J., Glazer, E. W., Mott, .'-I L. and Anderson, K. (1991) Modulation by tumor necrosis factor-a of human astroglial cell production of granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (GCSF). J. Neuroimmunol. 32, 269-278. Uzumaki, H., Okabe, T., Sasaki, N., Hagiwara, K., Takaku, F. and Itoh, S. (1988) Characterization of receptor for granulocyte colony-stimulating factor on human circulating neutrophils. Biochem.Biophys. Res. Commun. 156,1026-1032. Vadas, M. A., Nicola, N. A. and Metcalf, D. (1983) Activation of antibody-dependent cell-mediated cytotoxicity of human neutrophils and eosinophils by separate colony-stimulating factors. J. Immunol. 130,795-799. Vellenga, E., Rambaldi, A., Ernst, T. J., Ostapovicz, D. and Griffin, J. D. (1988) Independent regulation of M-CSF and G-CSF gene expression in human monocytes. Blood 71, 1529-1532. Wang, J. M., Chen, Z. G., Colella, S., Bonilla, M. A., Welte, K., Bordignon, C. and Mantovani, A. (1988) Chemotactic activity of recombinant human granulocyte colony-stimulating factor. Blood 72,1456-1460. Watari, K., Asano, S., Shirafuji, N., Kodo, H., Ozawa, K., Takaku, F. and Shin-ichi, K. (1989) Serum granulocyte colony-stimulating factor levels in healthy volunteers and patients with various disorders as estimated by enzyme immunoassay. Blood 73,117-122. Welte, K., Platzer, E., Lu, L., Gabrilove, J. L., Levi, E., Mertelsmann, R. and Moore, M. A. S. (1985) Purification and biochemical characterization of human pluripotent hematopoietic colony-stimulating factor. Proc. Nutl. Acad. Sci. USA 82,1526-1530. Yang, Y-C., Tsai, S., Wong, G. G. and Clark, S. C. (1988) Interleukin-1 regulation of hematopoietic growth factor production by human stromal fibroblasts. J. Cell. Physiol. 134,292-296. Yuo, A., Kitagawa, S., Ohsaka, A., Saito, M. and Takaku, F. (1990) Stimulation and priming of human neutrophils by granulocyte colony-stimulating factor and granulocytemacrophage colony-stimulating factor: Qualitative and quantitative differences. Biochem. Biophys. Res. Commun. 171,491-497. Ziltener, H. J., Clark-Lewis, I., Hood, L. E., Kent, S. 8.H. and Schrader, J. W. (1987) Antipeptide antibodies of predetermined specificity recognize and neutralize the bio-activity of the pan-specific hemopoietin interleukin-3. 1. lmmunol. 138,1099-1104. Zsebo, K. M., Cohen, A. M., Murdock, D. C., Boone, T. C., Inoue, H., Chazin, V. R., Hines, D. and Souza, L. M. (1986) Recombinant human granulocyte colony stimulating factor: Molecular and biological characterization. Immunobiol. 172, 175-1 84. Zsebo, K. M., Yuschenkoff, V. N., Schiffer, S., Chang, D., McCall, E., Dinarello, C. A., Brown, M. A., Altrock, B. and Bagby Jr., G. C. (1988) Vascular endothelial cells and granulopoiesis: Interleukin-1 stimulates release of G-CSF and GM-CSF. Blood 71,99-103.