(CANCER RESEARCH 49. 3955-3960. July 15. 1989]

Prevention of Doxorubicin-induced Hematotoxicity in Mice by Interleukin 1Deborah A. Eppstein,' Carole G. Kurahara, Nicholas A. Bruno, and Timothy G. Terrell

Institutes of Cancer and Developmental Biology, Bio-Organic Chemistry (D. A. E., C. G. A'., A1.A. B.J, and Toxicologie Sciences [T. (i. T.], Syntex Research, Palo Alto,

California 94304

ABSTRACT

Interleukin la and interleukin \ßinduce peripheral neutrophilia withstimulation of granulopoiesis in bone marrow. The continuous administration of interleukin 1 (100 ng/day) to mice for 7 days by s.c.-implantedAlzet osmotic minipumps induced marked stimulation of granulopoiesisin marrow and spleen in normal mice, and protected against the markeddepletion of myeloid and erythroid cells in bone marrow of mice treatedwith single injections of either 20 or 30 mg/kg doxorubicin (DXN).Interleukin 10 infusion also protected against DXN-induced atrophy ofthymus and secondary lymphoid organs. Single i.p. injection of eitherinterleukin la or interleukin ißat doses up to 1000 ng 24 h prior totreatment with DXN did not protect against the hematopoietic andlymphoid toxicities of DXN.

INTRODUCTION

Chemotherapy (1) as well as radiotherapy (2) can result insignificant depression of the lymphoid and hematopoietic systems of treated patients. Granulocytopenia is an importantcomplication of therapy and a major cause of death in cancerpatients (3). Consequently, it would be extremely beneficial tofind agents that would protect hematopoietic and lymphoidorgans from the suppressive effects of chemotherapeutic andradiation treatment regimens. Cytokines which act either directly or indirectly to regulate hematopoiesis offer utility inprotecting the hematopoietic system from cytotoxic treatmentsaimed at destroying cancer cells. Neta et al. (4, 5) found thatmurine or human IL-12 pretreatment of mice resulted in pro

tection from death by otherwise lethal doses of radiation,whereas neither recombinant murine interferon-7, recombinantmurine GM-CSF, nor recombinant lL-2 offered significantradioprotection at the doses and schedules tested. Morrissey etal. (6) observed that sublethally irradiated mice treated thera-peutically with IL-1/3 (injected i.p. twice daily for 20 days)responded with an increase in GM-CSF-responsive colonyforming cells in the marrow, followed by enhanced recoveryfrom the radiation-induced neutropenia.

With the background of IL-1 as a radioprotector in vivo (4-6), we wished to determine if IL-1 could also protect from themyelosuppressive effects of chemotherapy. We focused on themyelosuppressive effects of DXN (Adriamycin), as DXN is oneof the most widely used cancer chemotherapeutic drugs whichhas myelotoxicity as its acute dose-limiting toxicity (1). Themouse has been shown to be a clinically predictive model forstudying the myelosuppressive effects of several cytotoxic drugsincluding DXN (7). We now demonstrate that continuous s.c.infusion of IL-1 (a or ,>>')has significant activity in protecting

mice from the hematopoietic and lymphoid toxicities of DXN,whereas a single i.p. injection 24 h prior to DXN was ineffective.Analysis of the effects of IL-1 on hematopoietic and lymphoid

Received 10/19/88; revised 3/15/89: accepted 4/12/89.The costs of publication of this article were defrayed in part by the payment

of page charges. This article must therefore be hereby marked advertisement inaccordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1To whom requests for reprints should be addressed, at Syntex Corporation.3401 Hillview Avenue. Palo Alto. CA 94303.

! The abbreviations used are: IL-1. interleukin 1; GM-CSF, granulocyte-macrophage colony stimulating factor; DXN. doxorubicin: IL-2. interleukin 2;EMH. extramedullary hematopoiesis.

tissues of normal mice provided a basis for understanding itschemoprotective effects on these organs. Along analogous lines,Moore and Warren (8) have recently demonstrated that micetreated with 5-fluorouracil showed enhanced myelopoietic regeneration after i.p. treatment with IL-1«twice daily for 10days. Castelli et al. (9) have found that i.v. administration ofIL-la, either as a single injection 20 h preceding or as dailyinjections from 2-7 days postlethal dose of cyclophosphamide,

protected mice from death. A concomitant increase in colonyforming unit cultures as well as increased cellularity in bonemarrow and peripheral blood was observed.

MATERIALS AND METHODS

Mice. Female C57BL/6J mice (16-18 g) were purchased from TheJackson Laboratory. Bar Harbor, ME.

Reagents and Treatments. Doxorubicin hydrochloride (DXN; Adriamycin) was purchased from Sigma Chemical Co.. St. Louis, MO, orfrom Adria Labs, Inc., Columbus, OH. DXN was dissolved in 0.85%NaCl solution and administered to mice as a single (20 or 30 mg/kg)i.p. injection. Recombinant human IL-1«and IL-lfi were obtained fromImmunex, Seattle, WA, and were determined to contain 40 pg endo-toxin//jg protein and 3 pg endotoxin/^g protein, respectively, by theLimulus amebocyte lysate test. The IL-In and IL-Iff preparations eachcontained approximately 2 x 10* thymocyte units IL-1 activity/mg

protein, where one unit results in 50% of maximum proliferation ofthymocytes obtained from 5- to 8-week-old C3H-HEJ mice (10). Inter-leukin-ltt and IL-1/3 were prepared in 0.85% NaCl solution (with 0.1

mg/ml mouse serum albumin added for stability). Doses of 1, 10. 100,or 1000 ng/day for 7 days were infused s.c. into the mice via osmoticminipumps (Model 2001; Alza Corp., Palo Alto, CA), or as a singlei.p. dose of 100 or 1000 ng/day. All treatments with IL-1«or IL-1/3were initiated 24 h prior to DXN treatments.

Pathological Evaluations. Blood samples were collected by cardiocen-tesis while the mice were under halothane anesthesia. Euthanasia wasperformed by cervical dislocation and necropsy examinations wereperformed immediately after death. Tissues were preserved in neutral10% formalin and processed for light microscopy by standard procedures. Sections were stained with hematoxylin & eosin. Tissues examined included sternum for section of bone marrow, spleen, thymus,mesenteric lymph node, liver, heart, kidneys, and lung. Bone marrowsmears were prepared at necropsy, fixed in methanol and stained withWright Giemsa stain.

Sections of bone marrow and spleen were graded histologically forthe degree of hematopoietic activity. Bone marrow sections from treatedanimals were compared to sections from control animals and therelative increase or decrease in granulopoiesis, erythropoiesis or numbers of megakaryocytes was scored as follows: —¿�,no change from

control; ±.minimal increase or decrease in some but not all animals inthe group; + to +++++, relative values assigned by visual inspectionfor minimal to severe increase or decrease compared to controls. Forincreased (>) cellularity, + = 10-20% increase over control; ++ = 2xincrease; +++ = 3x increase; ++++ = 4x increase. For decreased (<)cellularity, + = 80% of control; ++ = 50% of control; +++ = 20% ofcontrol; ++++ = 10% of control; +++++ = less than 5% of control.Spleen sections of control mice had little evidence of EMH. Spleensections from treated mice were scored for increases in extramedullarygranulopoiesis or erythropoiesis and numbers of megakaryocytes ascompared to controls. The relative values for the grades used for spleensections were: + = 2x increase; ++ = 5x increase; +++ = lOx increase;++++ = 20x increase; +++++ = 30x increase compared to controls.

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INTERLEUKIN I AS A HEMATOPROTECTOR

In addition, lymphoid atrophy or hyperplasia in sections of thymus,spleen, and lymph nodes was scored for severity with (+ to +++++),minimal to severe, respectively, based on histolopathological criteriafor these changes.

Hematology measurements were performed on an ELT-8 Hematol-ogy Analyzer (Ortho Instruments, Westwood, MA). Differential leukocyte counts were determined by visual appraisal of Wright Giemsa-stained blood smears.

RESULTS

Dose Response. The effects of graded doses of IL-1/3 deliveredby continuous 7-day s.c. infusion on peripheral blood differential counts are illustrated in Fig. 1. Interleukin-1/3 induced arelative peripheral neutrophilia and lymphopenia at a dose levelas low as 100 ng/day, with a dose-dependent increase at 1000ng/day. A slight increase in percentages of band neutrophilswas observed at all doses tested.

Histopathological changes in hematopoietic and lymphoidtissues are shown in Table 1. The response to IL-1/3 in the bonemarrow was primarily stimulation of granulopoiesis at dosesthat produced changes in the peripheral blood, as evidenced byincreased numbers of myeloid precursor cells. Increased EMH,predominantly granulopoiesis, was observed in spleen with adose-related increase in severity. Megakaryocytes were alsoincreased in bone marrow and spleen. Slight stimulation oferythropoiesis was observed in the spleen; however, a minimaldecrease in erythroid precursor cells was observed in the bonemarrow of mice receiving IL-1/3 at 1000 ng/day.

The IL-1/3 infusion at 1000 ng/kg produced mild thymicatrophy characterized by depletion of cortical thymic lymphocytes. Lymphoid hyperplasia was also observed in lymph nodesat that dose.

Necrosis and inflammation were observed in the skin overlying the drug delivery portal of the osmotic pumps in micereceiving 1000 ng/day. Therefore, a dose of 100 ng/day wasselected as the optimal IL-1/3 dose which produced stimulationof hematopoietic tissues without producing unacceptable tox-icity, and that dose was used in subsequent studies.

Comparison of Single Injection and Infusion of IL-la and II .-1/8. The hematological (Table 2) and morphological (Table 3)changes were similar in mice treated with either IL-la or IL-1/3by continuous infusion for 7 days. In both groups there wasan absolute neutrophilia with lymphopenia, with no change in

Table 1 Effect of dose of continuous IL-Ißinfusion on hematopoietic andlymphoid tissues in vivo

10 ng/day 100 ng/day 1000 ng/day

NELrrnopHiLS LYMPHOCYTES

Fig. I. I Mivi of dose of continuous 111.; infusion on differential leukocytevalues. C57B1/6J mice (A7= 5/group) received continuous infusion of IL-l/i atthe doses indicated from s.c.-implanted osmotic minipumps at rate of 8 >J/h for7 days. Blood for hematology collected on Day 7. Differential leukocyte countrecorded as mean percentage ±SE for each treatment group. Statistical significance compared to the control group; *. P < 0.05: **. P < 0.01.

Dose of IL-lri(ng/day)°

Tissue/lesion IO 100 1000

Bone marrow>Granulopoiesis/>

<Erythropoiesis>Megakaryocytes

Spleen>Granulopoiesis>Erythropoiesis>Megakaryocytes

ThymusAtrophy

Lymph nodesHyperplasia NEá NE NE

" C57BI/6J mice (n = 5/group) received continuous infusion of IL-lff froms.c.-implanted osmotic minipumps at a rate of 8 nl/h for 7 days. The controlgroup animals received 0.9% NaCI solution.

* Symbols: >, increased; <, decreased.' Histopathological severity scores: -, not present; ±,minimal in some, but

not all, of the mice in the group. Bone marrow increased (>) cellularity: + = 10-20% increase; ++ = 2x control; +++ = 3x control; ++++ = 4x control; +++++= 5x control. Bone marrow decreased (<) cellularity: + = 80% of control; ++ =50% of control; +++ = 20% of control; ++++ = 10% of control; +++++ = lessthan 5% of control; Spleen increased (>) cellularity: + = 2x control; ++ = Sxcontrol; +++ = lOx control; ++++ = 20x control; +++++ = 30x control;Thymus and lymph node, + to +++++ = minimal to severe, respectively.

d NE, not examined.

total leukocyte values. Erythrocyte counts, hematocrits, andhemoglobin values were slightly decreased, and platelet countswere increased. These changes in the peripheral blood wereaccompanied by an increase in granulopoiesis in bone marrow(Fig. 2), spleen, and liver, and a slight suppression of erythropoiesis in bone marrow. A slight stimulation of erythropoiesiswas evident in the spleen. Megakaryocytes were increased inbone marrow and spleen. Minimal thymic atrophy was observedwith both IL-la and IL-1/3 infusion.

Mice that received a single i.p. injection of 100 ng of eitherIL-la or IL-1/3 had no changes in hematological parameters,and only minimal stimulation of bone marrow granulopoiesiswhen examined 7 days after injection. At 1000 ng by single i.p.injection, minimal stimulation of both granulopoiesis anderythropoiesis was observed in bone marrow and spleen withboth IL-la and IL-1/3. No effect was observed in the thymuswith a single dose of either interleukin.

Moderate lymphoid hyperplasia was observed in lymph nodesand spleen of mice treated with both IL-la and IL-1/3, whetherby infusion or a single i.p. injection.

Effect of Infusion of IL-la or IL-1/3 on Hematopoietic Toxicityof Doxorubicin. DXN was administered at two doses: 20 mg/kg, i.p., which approximates the LD50 value for these mice, or30 mg/kg, i.p. (Table 4). At 20 mg/kg, DXN produced mildstimulation of granulopoiesis in bone marrow but not spleenby Day 7. The degree of granulopoiesis in bone marrow wasmarkedly enhanced in mice receiving IL-la or IL-1/3, 100 ng/day for 7 days by s.c. infusion beginning 24 h prior to administration of DXN, and was increased over that observed witheither DXN or IL-1 treatment alone. The bone marrow changeswere accompanied by a neutrophilia and lymphopenia in theperipheral blood which was comparable to that observed inmice receiving infusion of IL-1 only (data not presented). Micereceiving IL-la in addition to DXN (20 mg/kg) had a markeddecrease in erythropoiesis in bone marrow, which was notobserved with DXN alone or IL-1/3 plus DXN. A mild increasein EMH was observed in the spleens of mice receiving theinfusion of IL-1/3 in addition to DXN, but it was not comparable

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INTERLEUKIN I AS A HEMATOPROTECTOR

Table 2 Comparison of effect of single injection versus continuous infusion oflL-l/i on nemalological parameters in miceTreatment"

IL-laParameter*Total

leukocytes(cells/nl)Neutrophils(cells/pl)Lymphocytes(cells/fil)Erythrocytes

(xlO6cells//J)Hemoglobin

(g/dl)Hematocrit(%)MCV<fl)MCH(pg)MCHC(g/dl)Platelets

(xlO4 cells/jil)Control(»

=5)6020

+685513±1265324±6609.82

±0.2114.2±0.437.8

±1.038.2±0.214.5+0.237.8+0.5769±46100-ng

singlei.p.injection(n =4)5725+

1063650±1575048+9129.61

±0.1313.9±0.336.8

±0.538.2±0.614.4

±0.337.8±0.2873±32100

ng/days.c.infusion(n

=5)7300

+7984026±699r•¿�"2923+51"8.62±0.12"12.2

±0.2"31.8±1.0"36.8

±0.714.2+0.238.7+0.5918+

139IL-1/3100-ng

singlei.p.injection(n

=3)7500

±1416484+1176941

±15189.25±0.3012.7+0.634.7±1.237.7+0.313.8

±0.236.7±0.7641±43100

ng/days.c.infusion(1

=5)5230

±13872400±9892699±272"8.72±0.24"12.4

±0.3"32.0+1.2"36.6

±0.5'14.2+0.238.8±0.41173+ 50"

" Results of two different studies were combined. C57BI/6J mice received IL-ln or IL-lfi by a single i.p. injection at doses of 100 ng or by continuous infusionfrom s.c.-implanted osmotic minipumps at a dose of 100 ng/day for 7 days. Six animals were assigned to each treatment group except for the group receiving 111..at 100 ng, i.p. (n = 3); however some specimens were inadequate for analysis due to the presence of clots. Blood for hematology values was collected on Day 7.

* The abbreviations used are: MCV, mean corpuscular volume; MCH, mean corpuscular hemoglobin; MCHC. mean corpuscular hemoglobin concentration.c Statistical significance compared to the control group; " P < 0.01; ' P < 0.05.

Table 3 Comparison of effects oflL-Ia or IL-lßadministered by continuous infusion or single injection on hematopoietic and lymphoid tissues in miceIL-1«" IL-1/3

Tissue/lesionBone

marrow>Granulopoiesis*

>Erylhropoiesis<Erythropoiesis>MegakaryocytesSingle

i.p.injection(100ng)±cSingle

i.p.injection(lOOOng)±

+Continuous

s.c.infusion (100

ng/day)+++-M-+Single

i.p.injection(lOOng)+

±+Single

i.p.injection(lOOOng)±

+Continuous

s.c.infusion (100

ng/day)+++++±

Spleen>Granulopoiesis>Erythropoiesis>MegakaryocytesLymphoid hyperplasia

ThymusAtrophy

Lymph nodesHyperplasia

±++

++±

°Results of two different studies are combined. C57B1/6J mice received IL-ln or IL-1/3 by a single i.p. injection at doses of 100 ng (n = 6/group) or 1000 ng (n •¿�•3/group), or by continuous infusion from s.c.-implanted osmotic minipumps at a dose of 100 ng/day for 7 days (n = 6/group). Necropsy was on Day 7.

* Symbols: >, increased; <, decreased.' Histopathological severity scores same as in Table 1.

to that observed in mice treated with IL-/3 only (Table 3).At 30 mg/kg, DXN produced marked depletion of both

myeloid and erythroid cells in bone marrow (Fig. 2). Concurrentinfusion of IL-10 protected against the bone marrow effects,and resulted in increased granulopoiesis in the marrow comparable to that obtained with IL-1/3 alone (Fig. 2). Moderatestimulation of EMH was also observed in spleens of micetreated with the IL-1/3 in addition to DXN (30 mg/kg).

Thymic atrophy characterized by depletion of cortical thymiclymphocytes was produced by DXN at doses of 20 or 30 mg/kg. IL-1/3 partially protected mice from the thymic lymphoiddepletion, but no protection was observed with IL-la infusion.IL-1/3 also protected mice from atrophy of secondary lymphoidtissues of spleen and lymph node which was produced by DXNat 30 mg/kg.

There were no lesions present in other tissues, including liver,heart, kidney and lung, that were attributed to acute effects ofDXN.

Effect of Single Injection of IL-1 «or IL-1/3 on Hematotoxicityof Doxorubicin. The mice treated with a single injection of II-la or IL-1/3 24 h prior to DXN treatment had no evidence ofprotection from the hematotoxicity (Table 5). Mild to moderatestimulation of granulopoiesis in bone marrow was observed inmice receiving IL-1 prior to DXN at 20 mg/kg, which was

comparable to that observed in mice receiving DXN alone. At30 mg/kg DXN, with or without prior IL-1 treatment, therewas a marked decrease in both erythroid and myeloid precursorcells in marrow sections.

Thymic atrophy was severe in all groups, although partialprotection from the atrophie changes in secondary lymphoidtissues of spleen and lymph node was observed with both IL-la and IL-1/3 administered in single doses of 1000 or 100 ng,respectively.

DISCUSSION

Our results demonstrate that both II.-I« and IL-1/3 wheninfused via 7-day Alza minipumps produced comparable stimulation of hematopoiesis and lymphoid hyperplasia in mice.Such an IL-1 infusion initiated 1 day prior to doxorubicininjection protected mice from the severe myelotoxicity of DXNand resulted in overall increased hematopoiesis in bone marrowand spleen such as obtained in IL-1 only treated mice. It isnoteworthy that a single injection of IL-la or IL-1/3, given 24h prior to the DXN, did not protect from the DXN-inducedhematotoxicity, even when doses up to 1000 ng of IL-1 weregiven. This finding is quite different from the effects of IL-1 onradioprotection, in which a single injection of 100 or 200 ng

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INTERLEUKIN I AS A HEMATOPROTECTOR

£.>:•íj*̂ viC yi^î"

C aFig. 2. Photomicrographs of bone marrow of mice. H & E. x 300. a control animal infused with saline for 7 days. Marrow demonstrates normal cellularity and

approximate equal proportions of myeloid and erythroid precursor cells, b, animal infused with IL- \fì( 100 ng/day) via s.c. osmotic minipump for 7 days. This marrowhas a moderate increase in cellular density with increased myeloid cells and a slight reduction in erythroid cells, c. animal received DXN (30 mg/kg) i.p. 6 days priorto necropsy. Hypocellular marrow with distended, blood filled sinusoids, d, animal received IL-l^i (100 ng/day) via s.c. infusion for 7 days and DXN (30 mg/kg) i.p.on Day 2. This marrow has a marked increase in cellular density with a predominance of myeloid cells and very few erythroid cells present.

Table 4 Comparison of effect of continuous //,-/u or ¡l.-l,iinfusion on hematotoxicity ofdoxoruhicin in mice20 mg/kg" 30 mg/kg

Tissue/lesion DXN DXN -I- IL-1« DXN + IL-10 DXN DXN + IL-ld

Bone marrow>(ìranulopoiesis/>

<Granulopoiesis<Erythropoiesis>Megakaryocytcs

Spleen>Granulopoiesis>Erythropoiesis>MegakaryocytesLymphoid atrophy

ThymusAtrophy

Lymph nodesAtrophy NE' NE NE

" Single i.p. injection of DXN was given 24 h after implanting the osmotic minipumps. IL-1« or IL-lff was administered by continuous infusion at a dose of 100

ng/day for 7 days (n = 3/group).* Symbols: (>). increased: (<). decreased.' Histopathological severity scoressame as in Table 1.rfNE, not examined.

IL-1«or IL-l/tf, respectively, was sufficient to protect approxi- differences of these two forms of IL-1, but more extensivemately 80% of the mice from radiation-induced death (4). The studies would be needed to address this issue,occasional differences in responses to IL-1«and IL-l/j which In order to initially define how the IL-1 resulted in thiswe observed (Tables 3 and 4) may reflect pharmacokinetic hematopoietic protection, we studied the cellular changes in-

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INTERLEUKIN 1 AS A HEMATOPROTECTOR

Table 5 Effect of single injection ofIL-la or IL-Ißon hematotoxicity of doxorubicin in miceDXN (20 mg/kg)° DXN (30 mg/kg)

Tissue/lesion DXNDXN + IL-la

(lOOng)DXN + IL-10

(lOOng) DXN

DXN + 1L-1«(lOOng)

DXN + IL-1«(lOOOng)

DXN + IL-ltf(100 ng)

DXN + IL-10(lOOOng)

Bone marrow>Granulopoiesis*

<Granulopoiesis<Er>'thropoiesis

Spleen>GranulopoiesisLymphoid atrophy

ThymusAtrophy

Lymph nodesAtrophy

NE

NE

NE

NE"II Li or II I.; was administered by single i.p. injection 24 h prior to treatment with DXN by i.p. injection. Necropsy examination was 7 days following

doxorubicin treatment (n = 3/group).* Symbols: (>), increased; (<). decreased.' Histopathologic severity scores same as in Table 1.* NE, not examined.

duced in the various hematopoietic and lymphoid tissues. Theresults obtained with IL-1 alone correlated with the protectiveeffects observed. Thus 7-day s.c. infusion of IL-1 via minios-motic pumps resulted in stimulation of hematopoiesis by Day7. This was characterized by markedly increased granulopoiesisand slightly depressed erythropoiesis in bone marrow, accompanied by increased extramedullary hematopoiesis in spleenand liver. Although total WBC counts remained unchanged,the ratio of neutrophilia and lymphopenia following i.v. injection (11), which is accompanied by the selective release ofmature neutrophils from the marrow (12). Interleukin-1 alsoenhances granulopoiesis by inducing cytokines required formyeloid proliferation (13, 14). The mechanism for the depression of erythropoiesis with the IL-1 treatment is not known;however IL-1 has been shown to inhibit the action of erythro-poietin on erythroid precursors in the marrow (15).

We observed a slight atrophy of the thymus after 7-dayinfusion of either IL-la or IL-1/3 (Table 3); thymic atrophy wasalso observed by Morrissey et al. (6) after multiple daily injections of mice with IL-la. However, it is noteworthy that thesevere thymic atrophy induced by DXN treatment was partiallyreversed by the IL-1/3 infusion (Table 4). In contrast, Morrisseyobserved an increase in thymic cellularity with time after irradiation in control mice and no such increase in irradiated micereceiving IL-la treatment (6).

Mice receiving the 7-day infusion of IL-1 were protectedfrom the acute hematotoxicities of DXN alone. However, micereceiving a single injection of IL-la or IL-1/3 showed onlyminimal effects on hematopoietic and lymphoid tissues, andwere not protected from the hematotoxicity of DXN.

IL-1 has been shown to regulate the development of primitivehematopoietic cells in concert with lineage-specific colony stimulating factors (GM-CSF or CSF-1) (16). This activity waspreviously attributed to hemopoietin-1 (17), which now appearsto be the same as IL-1 (16). In light of the above, it is significantthat IL-1 also stimulates production of GM-CSF from vascularendothelial cells (18, 19) and from fibroblasts (20). GM-CSFstimulates granulocyte and monocyte colony formation (21)and thus probably plays a significant role in resistance toinfections (22). Continuous infusion of GM-CSF stimulatedhematopoiesis in monkeys (23) and hamsters (24), and shortened the neutropenic period in monkeys given autologous bonemarrow transplantation (25). Fourteen-day infusion of GM-CSF into AIDS patients resulted in an increase in circulating

neutrophils, eosinophils, and monocytes (26). Thus GM-CSFas a single agent may be useful in hematopoietic restoration.Furthermore, since IL-1 can synergize with GM-CSF in stimulating primitive hematopoietic cells (16), combination of thesetwo agents may be especially beneficial in hematopoietic regeneration. Moore and Warren (8) have recently shown synergy ofIL-1 and G-CSF in stimulating stem-cell recovery and hematopoietic regeneration in mice following treatment with 5-fluorouracil. Furthermore, the ability of IL-1 treatment toincrease platelets in peripheral blood (Table 2) and megakary-ocytes in bone marrow and spleen (Tables 1, 3, and 4) may beadditionally beneficial for use in conjunction with cancer chemotherapies which induce thrombocytopenia in addition to neutropenia. However, as IL-1 is also a mediator of inflammation(27), a cautionary note should be observed ¡nexploring itstherapeutic potential.

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