1 Pulmonary Section, The National Chuushin Matsumoto Hospital, Matsumoto 399-0021; 2 The First Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto 390-8621; and 3 Department of Respiratory Medicine, Graduate School of Medicine, Kyoto University, Kyoto 606-8057, Japan
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ABSTRACT |
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Cyclophosphamide is an alkylating antineoplastic agent used in several conditions. However, little is known about the mechanism of its pulmonary toxicity. In the present study, we determined that human lung fibroblasts release activity for neutrophils and monocytes in response to cyclophosphamide in a dose- and time-dependent manner. Checkerboard analysis revealed that both neutrophil and monocyte activities were chemotactic. The release of chemotactic activity was inhibited by lipoxygenase inhibitors and cycloheximide. Molecular-sieve column chromatography revealed that both neutrophil (NCA) and monocyte (MCA) chemotactic activities had multiple peaks. NCA was inhibited by a leukotriene B4 receptor antagonist and anti-interleukin-8 and anti-granulocyte colony-stimulating factor antibodies. MCA was attenuated by a leukotriene B4 receptor antagonist and anti-monocyte chemoattractant protein-1 and anti-granulocyte-macrophage colony-stimulating factor antibodies. The concentrations of interleukin-8, granulocyte colony-stimulating factor, monocyte chemoattractant protein-1, and granulocyte-macrophage colony-stimulating factor significantly increased in response to cyclophosphamide. These data suggest that lung fibroblasts may modulate inflammatory cell recruitment into the lung by releasing NCA and MCA in response to cyclophosphamide.
interstitial lung disease; interleukin-8; granulocyte colony-stimulating factor; monocyte chemoattractant protein-1; granulocyte-macrophage colony-stimulating factor
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INTRODUCTION |
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CYCLOPHOSPHAMIDE IS AN ALKYLATING AGENT used in several conditions including fibrosis associated with collagen vascular diseases and cryptogenic fibrosing alveolitis (11). A report (5) indicated that, paradoxically, cyclophosphamide may induce interstitial pulmonary inflammation and fibrosis. Cyclophosphamide has been associated with pulmonary toxicity in ~1% of the cases, manifested most frequently as interstitial pneumonitis and pulmonary fibrosis (5). However, little is known about the mechanism and risk factors of its pulmonary toxicity (5). Neither specific lung histopathological findings nor definite relationships with risk factors have been established (5, 18). Lung damage from cyclophosphamide can occur at any time after the start of treatment and is not dose dependent (24).
Sequestration of peripheral blood neutrophils and monocytes within the lung is characteristic of a number of acute and chronic pulmonary diseases (1, 10). The presence of neutrophils is determined by the local generation of chemotactic agents, which direct neutrophil migration from the vascular compartment to the alveolar space along chemotactic gradients. The alveolar macrophage is also derived predominantly from peripheral blood monocyte migration and, to a limited extent, from local replication (3). Although elicited neutrophils and macrophages serve a vital role in the host defense against a number of organisms, the presence of increased numbers of activated neutrophils and macrophages can lead to tissue injury via the excessive elaboration of inflammatory cytokines, proteolytic enzymes, and oxygen radicals (10, 23). Substantial investigation has focused on alveolar macrophages as a primary source of chemotactic factors (9). However, neutrophil (NCA) and monocyte (MCA) chemotactic activities have been found to be produced by endothelial cells (26), fibroblasts (27), and pulmonary epithelial cells (25).
The fibroblast is the principal cell of most connective tissues and is
involved in constituting collagenous and noncollagenous components of
the extracellular matrix. This synthetic activity serves an important
structural function by providing a frame network for organ integrity.
In addition to this traditionally accepted function, recent studies
(15-17, 22) have demonstrated that
fibroblasts not only serve to maintain the connective tissue but are
important participants in the orchestration of acute and chronic
inflammation. In this context, fibroblasts released monocyte
chemoattractant protein (MCP)-1, granulocyte-macrophage
colony-stimulating factor (GM-CSF), and transforming growth factor
(TGF)- in response to interleukin (IL)-1
, tumor necrosis
factor-
, and platelet-derived growth factor, suggesting the
contribution to certain disease states (15-17, 22).
Recently, Takamizawa et al. (28) reported that
bleomycin stimulates human lung fibroblasts (HLFs) to release NCA and
MCA including IL-8, granulocyte colony-stimulating factor (G-CSF),
MCP-1, and GM-CSF. Therefore, the fibroblast, because of its anatomic
location, is in a pivotal position to participate in and direct
bidirectional communication between interstitial and vascular events in
drug-induced pulmonary inflammation and fibrosis.
Although airway epithelial cells and alveolar macrophages may play a role in inflammatory cell migration from the interstitium to the alveolar and bronchial spaces in response to cyclophosphamide, the underlying mechanism of inflammatory cell migration from the vascular compartment to the interstitium remains to be elucidated. The role of HLFs in inflammatory cell recruitment from the vascular compartment to the interstitium in response to cyclophosphamide is uncertain. The purpose of the present investigation was to determine whether HLFs could participate in the recruitment of inflammatory cells into the lungs. Specifically, the possibility of HLFs to release NCA and MCA in response to cyclophosphamide was evaluated. The results demonstrate that HLFs can release NCA and MCA, including IL-8, G-CSF, MCP-1, and GM-CSF, in response to cyclophosphamide.
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MATERIALS AND METHODS |
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Culture of HLFs. We used fetal HLFs (lung, diploid, human, passage 27), which are an established cell line and commercially available (American Type Culture Collection, Manassas, VA). HLFs were suspended at 1.0 × 106 cells/ml in F-12 medium supplemented with penicillin (50 U/ml; GIBCO BRL, Life Technologies, Grand Island, NY), streptomycin (50 µg/ml; GIBCO BRL), Fungizone (2 µg/ml; GIBCO BRL), and 10% fetal calf serum (GIBCO BRL). HLF suspensions (3 ml) were added to a 30-mm-diameter tissue culture dish (Corning, Corning, NY) and were cultured at 37°C in a 5% CO2 atmosphere. After 4-6 days in culture, the cells had reached confluence, and then the culture medium was replaced with 2 ml of medium supplemented as above and incubated for 1 more day.
Exposure of HLFs to cyclophosphamide.
The medium was removed from the cells by two washes with serum-free
F-12 medium, and the cells were incubated in the presence and absence
of cyclophosphamide. To determine the dose- and time-dependent release
of NCA and MCA, the cultures were incubated with various concentrations
of cyclophosphamide (0, 1.0, 10, and 100 µg/ml; Sigma, St. Louis, MO)
for 12, 24, 48, and 72 h at 37°C in a humidified 5%
CO2 atmosphere. Cyclophosphamide (10 µg/ml) did not cause
HLF injury (no deformity of cell shape, no detachment from the tissue culture dish, and >95% of cells viable by trypan blue exclusion) after 72 h of incubation. However, cyclophosphamide at 100 µg/ml caused substantial HLF cytotoxicity after 24 h of incubation. The
supernatant fluids were then harvested and stored at 80°C until
assayed. At least six separate HLF supernatant fluids were harvested
for each experimental condition.
Measurement of NCA and MCA. Polymorphonuclear leukocytes were purified from heparinized normal human blood with the method of Boyum (4). The resulting cell pellet, as determined by trypan blue and erythrosin exclusion, consisted of >96% neutrophils and >98% viable cells. The cells were suspended in Gey's balanced salt solution (GIBCO BRL) containing 2% bovine serum albumin (Sigma), pH 7.2, at a final concentration of 3.0 × 106 cells/ml. This suspension was used for the neutrophil chemotaxis assay.
Mononuclear cells for the chemotaxis assay were obtained from normal human volunteers, and red blood cells and neutrophils were separated from mononuclear cells by Ficoll-Hypaque density centrifugation (28). The preparation routinely consisted of 30% large monocytes and 70% small lymphocytes as determined by morphology andMolecular-sieve column chromatography of the chemotactic activity. To determine the approximate molecular mass of the released chemotactic activity in the supernatant fluids harvested at 72 h in response to 10 µg/ml of cyclophosphamide, molecular-sieve column chromatography was performed with Sephadex G-200 (Pharmacia, Piscataway, NJ). At a flow rate of 6 ml/h, the HLF culture supernatant fluid was eluted with PBS, and fractions after the void volume were evaluated for NCA and MCA in duplicate.
Effects of metabolic inhibitors on the release of NCA and MCA. The effects of the nonspecific lipoxygenase inhibitors nordihydroguaiaretic acid (NDGA; 100 µM; Sigma) and diethylcarbamazine (DEC; 1 mM; Sigma) and the 5-lipoxygenase inhibitor AA-861 (100 µM; Takeda Pharmaceutical, Tokyo, Japan) on the release of NCA and MCA were evaluated in response to 10 µg/ml of cyclophosphamide after 72 h of incubation. To further examine the involvement of protein synthesis in the release of chemotactic activity, cycloheximide (20 µg/ml; Sigma) was added to inhibit protein synthesis. At these concentrations, NDGA, DEC, and AA-861 inhibited the release of leukotriene (LT) B4 in HLF cultures (14) and did not cause cytotoxicity to HLFs after 72 h of incubation.
Effects of a LTB4 receptor antagonist
on NCA and MCA.
Because release of NCA and MCA was blocked by 5-lipoxygenase inhibitors
and because both NCA and MCA were extracted into ethyl acetate, a
LTB4 receptor antagonist (ONO-4057; ONO Pharmaceuticals, Tokyo, Japan) at a concentration of 105 M was used to
evaluate the involvement of LTB4 as NCA and MCA in the
crude supernatant fluids and in the column chromatography-separated lowest molecular mass fractions.
Measurement of LTB4 in the supernatant fluids. The concentration of LTB4 in the supernatant fluids was measured with a radioimmunoassay as previously described (21).
Effects of polyclonal antibodies to IL-8,
G-CSF, MCP-1, GM-CSF,
TGF-, and regulated on activation normal
T cell expressed and secreted.
Because the effects of metabolic inhibitors suggested the involvement
of peptides as NCA and MCA, we assessed chemokines known as NCA and
MCA. The neutralizing antibodies to IL-8, G-CSF, MCP-1, GM-CSF,
TGF-
, and regulated on activation normal T cell expressed and
secreted (RANTES; Genzyme, Cambridge, MA) were added to the HLF
supernatant fluids that were harvested at 72 h in response to 10 µg/ml of cyclophosphamide at the suggested concentrations to inhibit
these cytokines and were incubated for 30 min at 37°C. Then these
samples were used for chemotactic assay. It has been previously
reported (13, 16, 28) that these antibodies inhibited each
chemokine-induced NCA and MCA and each antibody did not influence the
neutrophil and monocyte chemotaxis induced by activated serum or
formyl-methionyl-leucyl-phenylalanine. As a negative control, we used
nonimmune IgG; nonimmune IgG did not have any influence on
cyclophosphamide-conditioned medium.
Measurement of IL-8, G-CSF,
MCP-1, GM-CSF, TGF-,
and RANTES in the supernatant fluids.
The concentrations of IL-8, G-CSF, MCP-1, GM-CSF, TGF-
, and RANTES
in HLF supernatant fluids cultured for 72 h with 10 µg/ml of
cyclophosphamide were measured by ELISA according to the
manufacturer's directions. GM-CSF and RANTES kits were purchased from
Amersham, and the minimum concentration detected by these methods was
2.00 pg/ml for GM-CSF and 15.6 pg/ml for RANTES. IL-8, MCP-1, and
TGF-
kits were purchased from R&D Systems (Minneapolis, MN), and the minimum detectable concentrations of IL-8, MCP-1, and TGF-
were 10.0 pg/ml, 31.3 pg/ml, and 0.31 ng/ml, respectively. G-CSF (CLEIA) kit was
obtained from Chugai Pharmaceutical (Tokyo, Japan), and the minimum
detectable concentration of G-CSF was 1.0 pg/ml.
Statistics. In experiments where multiple experiments were performed, differences between groups were tested for significance with one-way analysis of variance, with Fisher's multiple range test applied to data at specific time and dose points. In experiments where a single measurement was made, the differences between groups were tested for significance with Student's paired t-test. In all cases, a P value < 0.05 was considered significant. Data are expressed as means ± SE.
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RESULTS |
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Release of NCA and MCA from
HLFs.
HLFs released NCA and MCA in a dose-dependent manner in response to
cyclophosphamide (Fig. 1). The lowest
doses of cyclophosphamide to stimulate HLFs were 1 µg/ml for
neutrophils and 0.1 µg/ml for monocytes. Increasing concentrations of
cyclophosphamide up to 10 µg/ml progressively increased the release
of NCA and MCA. At a concentration of 100 µg/ml, NCA and MCA dropped
because cyclophosphamide caused cytotoxicity to HLFs after 24 h.
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Inhibition of the release of chemotactic activity by metabolic
inhibitors .
The supernatant fluids incubated with 10 µg/ml of cyclophosphamide in
the presence of NDGA, DEC, and AA-861 showed a decrease in release of
NCA and MCA (P < 0.01; Fig.
3). Cycloheximide inhibited the release
of both NCA and MCA (P < 0.001; Fig. 3). The
combination of NDGA and cycloheximide almost completely inhibited the
release of both NCA and MCA (P < 0.001; Fig. 3).
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Molecular-sieve column chromatography findings of the released
chemotactic activities.
Molecular-sieve column chromatography with Sephadex G-200 revealed that
NCA was heterogeneous in size (Fig.
4A). At least three peaks of
NCA with estimated molecular masses before and after cytochrome
c (molecular mass 12,300 Da) and an additional peak after
quinacrine (molecular mass 450 Da) were separated by column
chromatography. The released MCA was also heterogeneous (Fig.
4B). At least three peaks of MCA seemed to be separated by
column chromatography: two peaks with the estimated molecular masses
before cytochrome c and an additional peak after quinacrine.
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Inhibition of NCA and MCA by
LTB4 receptor antagonists.
Both NCA and MCA of the crude samples were significantly inhibited by
addition of the LTB4 receptor antagonist ONO-4057 (~70% for NCA and 40% for MCA; P < 0.01; Figs.
5 and 6).
ONO-4057 also inhibited the column chromatography-separated lowest
molecular mass peak of NCA and MCA (~80% for NCA and 60% for MCA).
The LTB4 receptor antagonist at concentrations of
105 to 10
9 M inhibited the neutrophil
migration dose dependently in response to 10
7 M
LTB4 but showed no inhibitory effects on
formyl-methionyl-leucyl-phenylalanine- and endotoxin-activated
serum-induced neutrophil and monocyte chemotaxis (data not shown).
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Release of LTB4 from HLFs. The measurement of LTB4 in the supernatant fluids by radioimmunoiassay revealed that HLFs released a significant amount of LTB4 in the baseline culture condition. However, the addition of cyclophosphamide at a concentration of 10 µg/ml for 72 h did not induce LTB4 release from HLFs [252 ± 20 (control) vs. 277 ± 6 (cyclophosphamide) pg/ml].
Inhibition of NCA and MCA by
polyclonal antibodies to IL-8, G-CSF,
MCP-1, GM-CSF, TGF-,
and RANTES.
Because HLFs had the potential to release chemokines and because
chemokines released from HLFs might be responsible for NCA and MCA, we
used polyclonal blocking antibodies to IL-8, G-CSF, MCP-1, GM-CSF,
TGF-
, and RANTES. Among these antibodies, anti-IL-8 and anti-G-CSF
antibodies inhibited NCA (P < 0.01; Fig. 5).
Anti-MCP-1 and anti-GM-CSF antibodies attenuated MCA (P < 0.001; Fig. 6). In contrast, anti-RANTES and anti-TGF-
antibodies
did not inhibit MCA. We evaluated the effect of anti-IL-8, anti-G-CSF,
anti-MCP-1, and anti-GM-CSF antibodies on the column
chromatography-separated NCA and MCA. These antibodies also inhibited
the chemotactic activities at the corresponding molecular mass
chemotactic peaks ~60-80%. Nonimmune IgG was used to evaluate
the effect of nonspecific antibody. Nonimmune IgG did not attenuate the
NCA and MCA in the same cyclophosphamide-conditioned medium.
The release of IL-8, MCP-1,
G-CSF, GM-CSF, TGF-,
and RANTES from HLFs with
cyclophosphamide.
The measurement of chemotactic cytokines by ELISA revealed that
cyclophosphamide at a concentration of 10 µg/ml for 72 h of incubation stimulated the release of IL-8 and G-CSF as NCA
(P < 0.001; Table 2) and
GM-CSF and MCP-1 as MCA (P < 0.05; Table 2).
In contrast, RANTES was not detected in HLF supernatant fluids. The
concentrations of cytokines from HLFs in response to cyclophosphamide also showed an increase in the corresponding molecular mass column fractions (Table 2).
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DISCUSSION |
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The incidence of pulmonary disease associated with cycloheximide is estimated to be <1%. Lung damage from cyclophosphamide can occur at any time after the start of treatment and is not dose dependent (5). Cases have been reported after administration of as little as 150 mg of cyclophosphamide (8), and the period between administration of the drug and the onset of symptoms has varied from 2 wk (6) to 13 yr (12). When the total dose does not exceed 10 g, regression is usually seen after withdrawal of the drug (18). However, the mechanism of pulmonary toxicity is uncertain. Allergic reaction was suggested by Batist and Andrews (2). Morse et al. (20) have shown that in mice, a single dose of cyclophosphamide (100 mg/kg) produced irreversible pulmonary fibrosis, characterized by an atypical histological picture, increased content of hydroxyproline in the lungs, and decreased lung volume and compliance. The changes were amplified in animals exposed to 70% oxygen. Two types of alveolar macrophages were demonstrated, and the authors suggested that the macrophages stimulate fibroblast proliferation. But the role of lung fibroblasts in inflammatory cell recruitment from the vascular compartment to the interstitium in response to cyclophosphamide is undetermined.
It has been reported (17, 28) that lung fibroblasts have
the potential to release IL-8 and G-CSF in response to tumor necrosis
factor-, IL-1
, or bleomycin. In the present study, the blocking
antibodies to IL-8 and G-CSF attenuated NCA similarly (~40%).
Cyclophosphamide significantly stimulated the release of IL-8 and G-CSF
from HLFs. Thus the release of IL-8 and G-CSF as NCA may at least
partly explain neutrophil infiltration to the interstitium in response
to cyclophosphamide.
Wang et al. (29) reported that the concentration of G-CSF as NCA was 7-70 ng/ml, although the concentration of G-CSF detected in the HLF supernatant fluids was less than that reported. We performed neutrophil chemotaxis by using human recombinant G-CSF. The chemotactic concentration of G-CSF as NCA was from 10 to 100 pg/ml (13). The discrepancies in G-CSF concentration as a neutrophil chemotactic factor may be due to the difference in neutrophil separation. Because the blocking antibodies to G-CSF inhibited both total NCA in the supernatant fluids and NCA in the column chromatography-separated peaks, the contribution of G-CSF to NCA may be as a direct chemoattractant rather than as the activation of neutrophils.
The identification of MCA released from HLFs is not complete. However, the inhibition of the release by cycloheximide treatment suggests that the activity is at least partly dependent on protein synthesis. MCA was attenuated by antibodies to MCP-1 and GM-CSF. The concentrations of MCP-1 and GM-CSF in the supernatant fluids reached those levels as MCA (28, 30). Thus HLFs released MCP-1 and GM-CSF at least partly as responsible MCA.
HLFs release MCP-1 and GM-CSF. However, the predominant MCA was GM-CSF
rather than MCP-1 and TGF-. GM-CSF is one of a group of
glycoproteins that have the ability to stimulate the in vitro proliferation and differentiation of macrophage progenitor cells (19). Thus the augmented release of GM-CSF from
fibroblasts in response to cyclophosphamide suggests that fibroblasts
may be profoundly concerned with macrophage recruitment,
differentiation, and proliferation in the lung interstitium rather than
in epithelial cells.
The inhibition of release by NDGA, DEC, and AA-861 suggests that the activity is composed of a lipoxygenase product. However, NDGA has the potential to inhibit the release of IL-8. NCA and MCA were inhibited by the LTB4 receptor antagonist. Although the release of LTB4 from HLFs in response to cyclophosphamide was not significant compared with the control value, the concentration of constitutively released LTB4 reached the chemotactic range of neutrophils and monocytes (14). Koyama et al. (14) previously reported that the LTB4 receptor antagonist and MCP-1 antibody inhibited the monocyte chemotactic response in the baseline condition and that HLFs constitutively release MCP-1 and LTB4. Thus it is no wonder that the combination of antibodies and LTB4 receptor antagonist inhibited the neutrophil and monocyte chemotactic responses less than the F-12 conditioned medium. Although the molecular-sieve column profiles did not show a striking increase in the lowest molecular mass peak in both neutrophil and monocyte migration, the LTB4 receptor antagonist abolished the lowest molecular mass chemotactic activity. Thus LTB4 may be one of the important chemoattractants released from fibroblasts constitutively for neutrophils and monocytes.
Cyclophosphamide stimulated the release of many cytokines from lung fibroblasts. The exact mechanisms for cyclophosphamide to stimulate fibroblasts, resulting in the release of cytokines, are uncertain, and the mechanism of activation or synthesis of 5-lipoxygenase in fibroblasts is also unclear. We speculate that the stimulatory potential of cyclophosphamide is not enough for the activation or synthesis of 5-lipoxygenase in fibroblasts compared with other stimuli that induced LTB4 release from fibroblasts. However, it might be possible that cyclophosphamide induced the release of 12-hydroxyeicosatetraenoic acid (HETE) and/or 15-HETE, which were NCA and MCA. 12-HETE and/or 15-HETE in combination with LTB4 may explain the augmentation of the lowest chemotactic peaks.
In conclusion, cyclophosphamide stimulated HLFs to release NCA and MCA. The released activities were chemotactic as determined by checkerboard analysis. The released NCA and MCA with cyclophosphamide were IL-8, G-CSF, MCP-1, GM-CSF, and LTB4. These results suggest that lung fibroblasts may play a role in the inflammatory cell recruitment by releasing chemotactic activity in response to cyclophosphamide.
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FOOTNOTES |
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Address for reprint requests and other correspondence: S. Koyama, Pulmonary Section, The National Chuushin Matsumoto Hospital, 811 Kotobuki Toyooka, Matsumoto 399-0021, Japan (E-mail: koyama{at}ka3.so-net.ne.jp).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Received 29 February 2000; accepted in final form 9 January 2001.
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REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1.
Adamson, IYR,
and
Bowden DH.
Role of monocytes and interstitial cells in the generation of alveolar macrophages. II. Kinetic studies after carbon loading.
Lab Invest
42:
518-524,
1980[ISI][Medline].
2.
Batist, G,
and
Andrews JL.
Pulmonary toxicity of antineoplastic drugs.
JAMA
246:
1449-1453,
1981[Abstract].
3.
Blusse van Oud Albas, A,
Mattie H,
and
van Furth R.
A quantitative evaluation of pulmonary macrophage kinetics.
Cell Tissue Kinet
16:
211-219,
1983[ISI][Medline].
4.
Boyum, A.
Isolation of mononuclear cells and granulocytes from human blood.
Scand J Clin Lab Invest
97:
77-89,
1968.
5.
Cooper, JAD,
White DA,
and
Matthay RA.
Drug-induced pulmonary disease. Part 1: cytotoxic drugs.
Am Rev Respir Dis
133:
321-340,
1986[ISI][Medline].
6.
Dohner, VA,
Ward HP,
and
Standord RE.
Alveolitis during procarbazine, vincristine and cyclophosphamide therapy.
Chest
62:
636-639,
1972[Medline].
7.
Falk, W,
Goodwin RH, Jr,
and
Leonard EJ.
A 48-well microchemotaxis assembly for rapid and accurate measurement of leukocyte migration.
J Immunol Methods
33:
239-247,
1980[ISI][Medline].
8.
Ginsberg, SJ,
and
Comis RL.
The pulmonary toxicity of antineoplastic agents.
Semin Oncol
9:
34-51,
1982[ISI][Medline].
9.
Hunninghake, GW,
Gadek JE,
Fales HM,
and
Crystal RG.
Human alveolar macrophage-derived chemotactic factor for neutrophils. Stimuli and partial characterization.
J Clin Invest
66:
473-483,
1980[ISI][Medline].
10.
Hunninghake, GW,
Garrett KC,
Richerson HB,
Fantone JC,
Ward PA,
Rennard SI,
Bitterman PB,
and
Crystal RG.
Pathogenesis of the granulomatous lung disease.
Am Rev Respir Dis
130:
476-496,
1984[ISI][Medline].
11.
Johnson, MA,
Kwan S,
Snell NJC,
Darbyshire JH,
and
Turner-Warwick M.
Randomised controlled trial comparing prednisolone in combination in cryptogenic fibrosis alveolitis.
Thorax
44:
280-288,
1989[Abstract].
12.
Karim, FWA,
Ayash RE,
Allam C,
and
Salem PA.
Pulmonary fibrosis after prolonged treatment with low-dose cyclophosphamide.
Oncology
40:
174-176,
1983[ISI][Medline].
13.
Koyama, S,
Sato E,
Masubuchi T,
Takamizawa A,
Kubo K,
Nagai S,
and
Izumi T.
Alveolar type II-like cells release G-CSF as neutrophil chemotactic activity.
Am J Physiol Lung Cell Mol Physiol
275:
L687-L693,
1998
14.
Koyama, S,
Sato E,
Masubuchi T,
Takamizawa A,
Nomura H,
Kubo K,
Nagai S,
and
Izumi T.
Human lung fibroblasts release chemokinetic activity for monocytes constitutively.
Am J Physiol Lung Cell Mol Physiol
275:
L223-L230,
1998
15.
Larsen, CG,
Zachariae CO,
Oppenheim JJ,
and
Matsushima K.
Production of monocyte chemotactic and activating factor (MCAF) by human dermal fibroblasts in response to interleukin 1 or tumor necrosis factor.
Biochem Biophys Res Commun
160:
1403-1408,
1989[ISI][Medline].
16.
Le, J,
Weinstein D,
Gubler V,
and
Vilcek J.
Induction of membrane associated interleukin-1 by tumor necrosis factor in human fibroblasts.
J Immunol
138:
2137-2142,
1987
17.
Leizer, T,
Cebon J,
Layton JE,
and
Hamilton JA.
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,
1990[Abstract].
18.
Mark, JG,
Lehimgar-Zadeh A,
and
Ragsdale BD.
Cyclophosphamide pneumonitis.
Thorax
33:
89-93,
1978[Abstract].
19.
Metcalf, D.
The granulocyte-macrophage colony-stimulating factors.
Science
229:
16-22,
1985[ISI][Medline].
20.
Morse, CC,
Sigler C,
Lock D,
Hakkinen PJ,
Haschek WM,
and
Witschi HP.
Pulmonary toxicity of cyclophosphamide: a 1-year study.
Exp Mol Pathol
42:
251-260,
1985[ISI][Medline].
21.
Poubelle, PE,
Borgeat P,
and
Rola-Pleszczynski M.
Assessment of leukotriene B4 synthesis in human lymphocytes by using high performance liquid chromatography and radioimmunoassay methods.
J Immunol
139:
1273-1277,
1987
22.
Rollins, BJ,
Stier P,
Ernst T,
and
Wong GG.
The human homolog of the JE gene encodes a monocyte secretory protein.
Mol Cell Biol
9:
4689-4695,
1989.
23.
Sibille, Y,
and
Reynolds HY.
Macrophages and polymorphonuclear neutrophils in lung defense and injury.
Am Rev Respir Dis
141:
471-501,
1990[ISI][Medline].
24.
Spector, JI,
Zimbler H,
and
Ross JS.
Early-onset cyclophosphamide-induced interstitial pneumonitis.
JAMA
242:
2852-2854,
1979[Abstract].
25.
Standiford, TJ,
Kunkel SL,
Basha MB,
Chensue WS,
Lynch JP,
Toews GB,
Westwick J,
and
Strieter RM.
Interleukin-8 gene expression by a pulmonary epithelial cells. A model for cytokine networks in the lung.
J Clin Invest
86:
1945-1953,
1990[ISI][Medline].
26.
Strieter, RM,
Kunkel SL,
Showell HJ,
Remick DG,
Phan SH,
Ward PA,
and
Marks RM.
Endothelial cell gene expression of a neutrophil chemotactic factor by TNF-alpha, LPS, and IL-1 beta.
Science
243:
1467-1469,
1989[ISI][Medline].
27.
Strieter, RM,
Showell HJ,
Remnick DG,
Lynch JP, III,
Genord M,
Raiford C,
Eskandari M,
Marjs RM,
and
Kunkel SL.
Monokine-induced neutrophil chemotactic factor gene expression in human fibroblasts.
J Biol Chem
264:
10621-10626,
1989
28.
Takamizawa, A,
Koyama S,
Sato E,
Masubuchi T,
Kubo K,
Sekiguchi M,
Nagai S,
and
Izumi T.
Bleomycin stimulates human lung fibroblasts to release neutrophil and monocyte chemotactic activity.
J Immunol
162:
6200-6208,
1999
29.
Wang, JM,
Chen ZG,
Colella S,
Bonilla MA,
Welte K,
Bordignon C,
and
Mantovani A.
Chemotactic activity of recombinant human granulocyte colony-stimulating factor.
Blood
72:
1456-1460,
1988[Abstract].
30.
Wang, JM,
Colella S,
Allavena P,
and
Mantovani A.
Chemotactic activity of human recombinant granulocyte-macrophage colony-stimulating factor.
Immunology
60:
439-444,
1987[ISI][Medline].
31.
Zigmond, SH,
and
Hirsch JG.
Leukocyte locomotion and chemotaxis. New methods for evaluation, and demonstration of a cell-derived chemotactic factor.
J Exp Med
137:
387-410,
1973[ISI][Medline].