Accumulation of Polymorphonuclear Leukocytesin ReperfusedIschemic Canine Myocardium:Relation with Tissue Viability Assessedby fluorine- 18-2-Deoxy glucose Uptake

he infiltration of polymorphonuclear leukocytesduring the process of myocardial necrosis has been welldocumented (1 ). More recently, ieukocytes have beensuspected to play an active role in acute myocardialischemia, especially after reperfusion (2,3). This hypothesis is supported by the fact that pharmacologicmanipulation of leukocyte function was shown to reduce the extent of ischemic myocardial injury (4—6).

Received Aug. 25, 1987; revision accepted July 22, 1988.For reprints contact: William Wijns, MD, Positron Emission

Tomography Laboratory, Avenue Hippocrate 55, Box 5550, B-1200Brussels,Belgium.

Presented in part at the 34th Annual Meeting ofthe Society ofNuclear Medicine, Toronto, 1987.

Capillary plugging by leukocytes may exacerbate tissueinjury indirectly by inducing edema and no-reflow (7).Direct effects are mediated through the release of freeradicals, leukotnenes and proteolytic enzymes (8—10).Margination of leukocytes in response to chemotaxisand their activation are energy dependent processesinvolving stimulation of glucose metabolism via thehexose monophosphate shunt and to a lesser extent, viathe glycolytic pathway (10,11).

The role of leukocytes in reperfused myocardiumand their dependence on glucose metabolism have tobe recognized when glucose uptake is studied with theglucose analog, Fluorine-l8-2-deoxyglucose (FDG) andpositron emission tomography, in order to evaluatetissue viability. It has indeed been shown that ischemic

1826 Wijns,Melin,Lenerset al The Journalof Nudear Medicine

Accumulation of PolymorphonuclearLeukocytes in Reperfused Ischemic CanineMyocardium: Relation with Tissue ViabilityAssessed by fluorine- 18-2-Deoxyglucose UptakeWilliam Wijns, Jacques A. Melin, Norbert Leners, Augustin Ferrant, AndréKeyeux,Jacques Rahier, Michel Cogneau, Christian Michel, Anne Bol, Annie Robert,Hubert Pouleur, AndréCharlier, and Christian Beckers

Center ofNuclear Medicine, the Positron Emission Tomography Laboratory, the Division ofCardiology and the Department ofPathology, University ofLouvain, Brussels, Belgium

Polymorphonudear leukocytes may participate in reperfusion injury. Whether leukocytesaffect viable or only irreversibly injured tissue is not known. Therefore, we assessed the

accumulation of 111ln-labeled leukocytes in tissue samples characterized as either ischemicbut viableor necroticby metabolic,histochemical,andultrastructuralcriteria.Six open-chestdogs received left anterior descending coronary occlusion for 2 hr followed by 4 hrreperfusion. Myocardial blood flow was determined by microspheres and autologous 111Inlabeled leukocytes were injected intravenously. Fluorine-i 8-2-deoxyglucose, a tracer ofexogenousglucoseutilization,was injected3 hr after repertusion.Thedogswerekilled4 hrafter reperfusion.The riskand the necroticregionswere assessedfollowinginvivodyeinjectionandpostmortemtetrazoliumstaining.Myocardialsampleswereobtainedin theischemic but viable, necrotic and normal zones, and counted for 111lnand 18Factivity.Comparedto normal,leukocyteswere entrappedin necroticregions(111lnactMty: 207±73%)where glucoseuptakewas decreased(26 ±15%). A persistentglucoseuptake,markerofviability,was mainlyseen in riskregion(135 ±85%) where leukocytesaccumulationwasmoderate in comparison to normal zone (146 ±44%). Thus, the glucose uptake observed inviable tissue is mainly related to myocytes metabolism and not to leukocytes metabolism.

J Nucl Med 29:1826—1832,1988

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but viable tissue can be identified by a persistent orincreased FDG uptake (12,13).

The labeling of autologous canine ieukocytes withindium-i 11 (“In)oxine permits the quantification oftheir distribution in vivo during the inflammatory response. Experiments were designed in a canine modelofreperfused myocardial infarction in order to compare‘‘‘Inand ‘8Factivity in the same tissue samples char

acterized as either ischemic but viable or necrotic byhistochemical and uitrastructural criteria. The objectivewas to assess the extent ofleukocytes accumulation andto investigate whether they could significantly contribute to FDG uptake in reperfused myocardium.

METhODS

Animal PreparationSix mongrel dogs weighing 21 to 27 kg were studied after

an overnight fast. The dogs were anesthetized with sodiumpentobarbital (30 mg/kg), intubated and ventilated with roomair. Morphine sulfate (1 mg/kg intramuscularly) was given atinduction and throughout the remainder ofthe experiment tomaintain adequate anesthesia. After left thoracotomy, theheart was suspended in a pericardial cradle. Catheters, insertedinto the left atrium and descending aorta, were used forinjecting radioactive microspheres and recording arterial pressure. The dogs were subjected to an occlusion of the leftanterior descending coronary artery with an atraumatic vascular clamp.

Experimental ProtocolAll dogs were pretreated with lidocaine at the time of

coronary occlusion and prior to reflow. The experimentalprotocol is outlined in Figure 1. The left anterior descendingcoronary artery was occluded for 2 hr. Microspheres wereinjected into the left atrium at I 10 mm postocclusion. Tenminutes later, the occlusion was released. Autologous ‘‘‘Inlabeled leukocytes were injected intravenously either prior toocclusion (n = 3) or 2 hr after reperfusion. The reason for thedelay was it would allow for preparation of leukocytes in aclinical situation of reperfused infarction. Myocardial bloodflowwasdetermined 180mm after reperfusion.Thereafter,6to 8 mCi of FDG were injected intravenouslyand an arterialinput function was obtained by rapid arterial sampling. Theleft anterior descending artery was then occluded with a snare,blue dye injected into the left atrium as a marker for the

normal tissue, and the heart arrested with concentrated potassium chloride solution.

Afterdeath, the heartswere excised and the ventriclesweresectioned parallelto the atrioventriculargroove, forming fiveslices 1—1.5cm thick. Tissue specimens were immediatelyexcised with a scalpel for electron microscopy, from the centerand lateral portion of the area at risk and from the nonischemic region (stained by the dye). One endocardial sampleand one epicardialsampleweretaken from each of the threeregions. Thereafter, the basal surfaces of the heart slices andmarginsof the area at risk were traced onto acetate sheets.The slices were weighedand then placed in a solution oftriphenyl tetrazolium chloride(TTC) at 31C for 30 mm. AfterTTC staining, sampling sites previouslytaken for electronmicroscopy within the area at risk were identified as beingischemic (TTC+) or necrotic (TTC—).Endocardial and epicardialsampling for regional myocardialblood flow, for “Inand ‘8Factivity was made in the center of the infarct, inischemic but noninfarcted tissue, and in the nonischemicmuscle.Ischemicsampleswerecut in order to avoid contamination by nonischemic and necrotic tissue. Necrotic samplesweresubdividedinto homogeneous,confluentareasof necrosis and patchy necrosis. We called patchy necrosis islets ofunstained tissue intermixed with stained myocardium in suchway that homogeneous sampling would seem impossible.Normal endocardial (n = 40) and epicardial (n = 37) samplesweretaken foreach leftventricularring from remote tissueinthe center of the circumflex coronary artery distribution.Samples were weighed and counted in a scintillation wellcounterat appropriateenergywindows.

The size of myocardial infarcts and areas at risk weremeasured planimetrically. The fractions oftotal left ventricular area representingthe area at risk and the infarcted tissuewere multiplied by the weight of each left ventricularslice toobtain the percentage, by weight, of total left ventricle thatwas infarcted and at risk. Microspheres(15 ±l@zm)werelabeledwith “Srand @Nb.Myocardialblood flowwascalculated with the equation:

Qm = (Cm x R)/Cr

where Qm = myocardial blood flow (mI/mm); Cm = tissuecounts (counts/mm); R = reference arterial blood flow, andCr = counts in reference blood sample. To account for thedifferent scatter fractions and life times, all samples werecounted three times:on the day of the experiment,24 hr and4 wk later. A multichannel analyzer was used with the following windows:‘8F= 947 to 1097keV; ‘‘‘In:370 to 490 keY;

Sacrifice

In-IllLEUKO

n:34, FIGURE1

_J Expenmental protocol. LEUKO: leukocytes; LAD: left antetior descend

6 ing coronaryartery;FDG:18F-2-deoxyglucose.

0 2 4Hours

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Releaseocclusion

ISPHERES

4,

FDGSPHERES

LADocclusion

In-IllLEUKOI

n:3

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lschemicarea(TFC+)p ValuePatchyNecrosisp ValueNecrosis(TrC—)p

Valuenecrosisvs.

ISchemIC“1ln

activity(%)Injectionpriorto occlusion147 ±75

N.S.<0.02322±202

N.S.<0.02176 ±48

N.S.N.S.Injectionafterreperfusion145 ±39<0.05203 ±84N.S.255 ±97<0.03All146

±44<0.004254 ±347N.S.207 ±73<0.02No.ofdatapoints19712Endocardium569Epicardium1413

85Sr: 500 to 580 keY; 95Nb: 735 to 835 keY. A computerprogram was used to correct for activity overlap between theenergy windows. The rate of exogenous glucose utilizationwascalculatedapplyingthe FDG modeldescribedby Sokoloff(14). Fixed values for the four rate constants and a lumpedconstant of 0.67 were used to calculate the glucose metabolicrate in myocardial tissue (expressed in mg/min/lOOg)(15,16).

Samples obtained for electron microscopy were classifiedas showing either cell death or cell injury without definiteevidence of cell death. The criteria for injured cells weresubsarcolemmal blebs, separation and disruption of sarcomeres, and mitochondrial swelling. Criteria for cell death included amorphous matrix densities in mitochondria, disruption of the mitochondria, and marked clumping of nuclearchromatin along the nuclear membrane.

Preparation of Radiolabeled Leukocytes

Labeledleukocyteswerepreparedas follows.Venousblood(55 ml) was aspirated into a syringe containing 10 ml ACD.Forty fivemillilitersof the blood were mixed with 1.8 ml of2% methylcellulose in 0.9% sodium chloride and the red cellswere allowedto sediment for 45 mm. The remaining20 mlblood was centrifuged at 1,500 g for 10 mm to obtain cell freeplasma. The leukocyte-rich supernatant from the sedimentedblood was centrifuged at 80 g for 7 mm. The cell pellet wasthen resuspended in 4 ml of cell-free plasma. Granulocyteswere isolated by sedimentation on discontinuous densitygradient columns made up ofa mixture ofautologous plasmaand Percoll (17,18). Briefly, 50%, 60%, and 65% Percoll/plasma solutions were prepared and 2 ml of each were thenlayered into two test tubes, with 2 ml of the mixed leukocytesuspension on top of each. The gradients were spun at 200 gfor 15 mm and the pure granulocytes were recovered. Thecells were washed once in 0.9% sodium chloride and resuspended in 10 ml of 0.9% sodium chloride. After 15 mmincubation with [‘‘‘In]oxine,2 ml of cell-free plasma wereadded and the mixture was centrifugedat 80 g for 7 mm.After resuspensionin more plasma, the cellswere ready forinjection. The labeling efficiency was 91 ±5% and the meanactivity injected was 380 MCi.In vivo recovery of radioactivitywasassessedby calculatingthe percentofadministered radioactivity present in the circulation 30 mm after injection. Thepercent recovery of radiolabel was 35 ±6% (range 22—61%).Granulocytes were to some degree contaminated by red cells(14 ±2% of the final suspension and 4 ±1% of the injectedactivity) and by lymphocytes (10 ±2% ofthe cells in the final

suspension and 9 ±2% of the injected activity). No plateletswere left.

Data AnalysisMyocardial blood flow and tracer activity in ischemic and

necrotic samples were normalized by dividing these values bythe average activity in normal remote myocardium from thecorresponding left ventricular ring.

Results are expressed as absolute values or percent ofnormal and mean ±s.d. are given. The mean values of flowand tracer activity for each tissue type within each slice wereused as individual data points in the analysis of variance forall animals. This accounts for the differentnumber of samplesof each tissue type obtained in each animal. In Table 1,statistical analysis was performed by non parametric analysisof variance (Kruskal-Wallis test) with contrast (Mann-Whitney Wilcoxon test). In Table 2, a Wilcoxon signed rank

analysis was used to test the hypothesis that the median valuefor ischemic, patchy and necrotic tissue equals 100% of normal. A p value <0.05 was considered as significant.

RESULTS

Hemodynamic Data and Infarct SizeBy 110 mm postocclusion, at the time of the first

microsphere injection, mean heart rate was 135 ±18bpm, not different from control values (137 ±19 bpm)and mean aortic pressure was slightly decreased (1 14 ±32 mmHg), as compared to the preocclusion pressure(122 ±18 mmHg; not significant, N.S.).

Three hours after reperfusion, a small decrease inheart rate (123 ±32 bpm; N.S.) was observed and meanaortic pressure showed similar values than prior toocclusion (125 ±26 mmHg).

The risk/left ventricle area ratio was 22 ±3%. Therange of the infarct/risk region ratios was large. Onedog had no infarct at all; another dog had a 100%infarct; the other infarct/risk region ratios were 14, 22,26, and 28%. The data from all dogs were included inorder to study the entire spectrum of tissue injury.

Indium-ill Activity After ReperfusionThe normalized ‘‘‘Inactivities are given in Table 1.

The ‘‘‘Inactivity in the different tissue samples was not

TABLE 1Normalized 111lnActivity After Reperfusion

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lschemicarea(TIC +)Patchy necrosisNecrosis (TTC—)Myocardial

bloodflowDunngocclusion(%)38±l8@19@9t6@5tRange14—641

1—322—11Afterreperfusion(%)82 ±1986 ±4584 ±85Range48—11054—15216—206Metabolic

ratefor glucose(%)135 ±8535—2651

10±7518—18926

±15@4—69Range111ln

activity (%)146 ±44*254 ±347@207 ±73@Range94—200143—930126-324p

Values versus 100 % (normaltissue).a@ <@t

p < 0.001.

MBF(mI/min/100g)@

: ‘MR.Gk@

1.5

TABLE2Normalized Myocardial Blood Flow, Metabolic Rate for Glucose, and 111InActivity After Reperfusion

different between the dogs having received labeled leukocytes prior to occlusion (n = 40 data points) or afterreperfusion (n = 3 1 data points). For the subsequentresults presentation, the data have been pooled togetherbecause no differences in tracer concentrations wereobserved between the two groups ofdogs. As comparedto normal myocardium, ‘‘‘Inactivity was significantlyincreased in the reperfused zones, but with a gradientfrom the ischemic samples (146 ± 44%) to the patchy

(254 ±347%) and homogeneous necrosis (207 ±73%).The normalized ‘‘‘Inactivity was significantly differ

ent from 100% (normal area) in the ischemic samples(p < 0.01), mainly epicardial. Compared to normal andischemic tissue, the ‘‘‘Inactivity was significantly increased in necrotic samples, both for patchy (p < 0.004versus ischemic) and homogeneous necrosis (p < 0.02versus ischemic). On the other hand, there was nodifference between patchy and homogeneously necroticsamples, mainly endocardial.

Comparison of Blood Flow, Glucose Uptake,and “InActivity

Representative examples of distribution of myocardial blood flow during occlusion, metabolic rate forglucose and ‘‘‘Inactivity are given in Figures 2 to 5.The first example shows an unrolled cross section ofendocardial (Fig. 2) and epicardial (Fig. 3) samples in adog with endocardial necrosis in four samples (TTC—)within the risk region. In the necrotic endocardial sampies (Fig. 2), metabolic rate for glucose was decreasedand ‘‘‘Inactivity was increased by threefold. In theepicardial samples, without necrosis within the area atrisk (Fig. 3), two of the samples had an increasedmetabolic rate for glucose, whereas ‘‘‘Inactivity wasonly slightly increased. The second example is from adog without any necrosis as shown by the positive TTCstain inside the risk region (Figs. 4 and 5). In this case,viable endocardial tissue shows a rate of exogenous

ACTIVITY PROFILE OF AN UNROLLED CROSS SECTION

1@ f

@-.. MBF

@—[email protected] In-Ill

In-Ill(x103 cos.wts)

@111

9

7

5

2.0

70

60

50

ENDO#6054030

20

10

5

FIGURE2Relation between regional myocardialbloodflow(MBF)duringcoronaryocclusion, metabolic rate for glucose(MR-Glc) and 111lnactivity along anunrolled left ventricular endocardialcross-section from a representativedog. The four necrotic samples areunstainedby TTC(TTC—)withinthearea at risk delineated by the arrows.The foursamples inthe blueregionrepresent normal tissue.

S TIC- —t

t NonBlue 1-3I Blue 1Blue

1829Volume29 •Number11 •November1988

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ACTIVITY PROFILE OF AN UNROLLED CROSS SECTION

MBF(mI/min/ bOg)

MRGlcIn-Ill

(x103 counts)

11

9

7

5

70

60

50

40

30

20

10

5-C

EPI ‘60S

1.5

FIGURE 3Relationbetween myocardialbloodflow (MBF), metabolicrate for glucose(MR-Glc) and ‘11Inactivity alongan unrolledleft ventricularepicardialcross-sectionfrom the samedog asin Figure 2. All the samples werestained by TTC.

1-3T Blue 1Non Blue

1.Oj-‘ Blue ‘1'

..--. MBF

.—@ MR@Glc--- In-Ill

glucose utilization that is two times above remote tissuewhile ‘‘‘Inactivity is only 150% of normal tissue.

Table 2 shows the mean normalized values ofmyocardial blood flow (during occlusion and after reperfusion), the metabolic rate for glucose and the “Inactivity in the same tissue samples characterized byhistochemistry and electron microscopy. During occlusion, the decrease in myocardial blood flow was moreimportant in necrotic regions (patchy necrosis: 19 ±9%; homogeneous necrosis: 6 ±5%) than in ischemicregions (38 ±18% of normal samples). After reperfusion, the mean normalized flow was similar to the flowin normal tissue, but the large standard deviations innecrotic areas indicate a heterogeneity of flow in thesesamples. The metabolic rate for glucose was 2.0 ±1.1mg/min/lOO g in the ischemic samples which is not

different from the values measured in normal samples(3.3 ±2.2 mg/min/iOO g), showing a large interanimalvariation. Therefore, we divided the metabolic rates inischemic samples by the average value measured in thenormal samples from the corresponding left ventricularring. This normalized ratio was 135 ±85%. Two of thesix dogs had a glucose utilization rate over 150% ofnormal in the ischemic area. In the homogeneouslynecrotic samples, the metabolic rate for glucose was 26±15%, i.e., 0.7 ±0.6 mg/min/lOO g. This is significantly different from normal tissue (p < 0.001), ischemic samples (p < 0.001) and patchy necrosis (p <0.03). Thus, if we compare normalized ‘‘‘Inactivityand metabolic rate for glucose, ‘‘‘Inactivity was mainlyincreased in the necrotic regions where glucose metabolism was decreased. In the ischemic but viable area,

ACTIVITY PROFILE OF AN UNROLLED CROSS SECTION

MBF(mI/min/lOOg)

In-Ill(x i0@ counts)

14

3.5

3

2.5

2

FIGURE4Relation between regional myocardialbloodflow(MBF), metabolicrateforglucose(MR-GIc)and“1lnactivityalong an unrolled left ventilcular endocardial cross-section from a representativedog without myocardialnecrosis in the risk (nonblue) region.Note that the 111Inactivity scale isdifferent than in Figures 2 and 3.

30

20

10

Blue •‘t'Non blue t1}-1 Blue

MBF

MR@Gk

In-Ill

.—.

1830 Wijns,Melin,Lenerset al The Journalof NuclearMedicine

70

60

50ENDO.610

40

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5@

at NonBlue

ACTIVITY PROFILE OF AN UNROLLED CROSSSECTION

MBF(mI/min/lOOg)

70

where a normal or slight increase of metabolic rate wasfound, the increase of ‘‘‘Inactivity was only moderatecompared to the necrotic area.

DISCUSSION

The present study shows that polymorphonuclearleukocytes do accumulate in ischemic canine myocardium as early as 4 hr after reperfusion.

We used autologous ‘‘‘In-labeledleukocytes and direct tissue counting in a well counter for the quantification of the leukocytic infiltration. Initial studies withthis approach showed that maximal “Inuptake occurred in the subendocardium 72 hr after experimentalinfarction induced by permanent coronary occlusion(19-21). We confirm that reperfusionsignificantlyenhances the process of leukocytes accumulation (2,22). No difference was found when leukocytes wereinjected during occlusion or after reperfusion. Leukocyte entrapment within the necrotic area was more

important in the present study (ranging from 126 to930% of normal) than in the study by Engler et al.(77%—353%). This may be related to the fact that theydid not allow more than 5 mm of reperfusion beforedeath. In addition, we attempted to relate the entrapment ofleukocytes to the fate ofthe tissue at risk beingeither salvaged by reperfusion or eventually becomingnecrotic. Therefore, tissue sampling was guided by tetrazolium staining. This histochemical criterion wasconfirmed by electron microscopy showing no signs ofcell death in TTC stained myocardium (mainly epicardium). In this ischemic but viable tissue, ‘‘‘Inactivitywas only moderately increased to i46 ±44% of remotenormal myocardium. Conversely, in necrotic tissue(mainly endocardium) identified as patchy or homogeneous areas unstained by TTC, we observed a major

601-

Blue

In-Ill(x103 counts)

4

3.5

3

2.5

21@

50

40

EPI ‘610

4

330

20

102

FIGURE 5Relation between myocardial bloodflow (MBF), metabolicrate for glucose (MR-Glc)and 111Inactivity alongan unrolledleft ventricularepicardialcross-section from the same dog asin Figure 4.

but variable increase in ‘‘‘Inactivity ranging from 126%to 930% of normal. This wide range can be explainedby tissue heterogeneity. Areas of patchy necrosis obviously represent a mixture of tissue but even withinsamples designated as homogeneously necrotic by TTC,electron microscopy still revealed some cells withoutdefinite evidence of cellular death. The latter probablyrelates to the temporal heterogeneity ofischemic injury,not all cells suffering the same degree of injury at thesame time (23).

In this occlusion-reperfusion protocol, we comparedthe regional distribution of' ‘‘In-labeledleukocytes andFIX; uptake. This glucose analog has been used extensively with positron emission tomography to studymyocardial exogenous glucose utilization in vivo. Previous studies in a chronic dog model of reperfusedmyocardial infarction have indicated a shift in substratemetabolism toward preferential glucose utilization inischemic but viable myocardium (12,13). However, inorder to demonstrate that glucose uptake detected bypositron emission tomography reflects the metabolismof myocytes, we wanted to show that other cells are notdirectly involved in FDG uptake. Leukocytes were thetarget cells in this situation, because of their activeparticipation in the inflammatory response of acuteinfarction particularly after reperfusion and because oftheir dependence on glucose metabolism. The energyrequired to drive the contractile machinery of the neutrophils is supplied through hydrolysis of adenosinetriphosphate generated by anaerobic glycolysis. Also,the contact between leukocytes and chemotactic factorsresults in stimulation of the hexose monophosphateshunt and to a lesser extent, of the glycolytic pathway(24). Since FDG traces transmembranous exchange andphosphorylation ofglucose, increased flux through any

I Blue

MBFMR-Glc

In-Ill

1831Volume29 •Number11 •November1988

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of these two pathways from exogenous glucose wouldresult in increased FDG uptake. Supporting the hypothesis that leukocytes can accumulate FDG is the observation that the thoracotomy scar in chronic dogs occasionally shows significant FDG uptake on positronemission tomographic images (Wijns, Melin, et al: unpublished observations).

In summary, this study shows that polymorphonuclear leukocytes accumulate in necrotic tissue as earlyas 4 hr after reperfusion of ischemic canine myocardium. A persistent glucose uptake was mainly seen inischemic but viable tissue, where leukocytes accumulation was moderate in comparison to normal andnecrotic tissue. Conversely, little residual FDG uptakewas noted in necrotic tissue, where leukocytes accumulation was massive. Thus, these data provide indirectevidence that preserved FDG uptake is likely to reflectthe metabolism of viable myocytes. Direct evidenceconfirming these findings and further supporting theuse of these tracers in clinical imaging would be provided by microautoradiographic studies.

ACKNOWLEDGMENTS

This work was supported by Grants from the “Fondsde IaRecherche Scientifique Médicale―3-4540-85 and 3-4541-87.

Electron micrographs were prepared by M. Stevens fromthe Department of Pathology. The authors thank D. Ockrymovicz-Bemelmans and E. Willems for expert technical assistance as well as Henri Van Mechelen for his expert technicalassistance, and D. Vangeebergen for typing the manuscriptwith great care. The authors appreciate the generous supportof the Damman Foundation.

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3. Engler RL, Dahlgren MD, Morris DD, et al. Role ofleukocytes in response to acute myocardial ischemiaand reflowindogs.Am J Physiol1986;25l:H3l4-.H322.

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circulation. FedProc 1987; 46:2397—2401.8. Sacks T, Moldow FM, Craddock PR, et al. Oxygen

radicals mediate endothelial cell damage by complement stimulated granulocytes. J C/in Invest 1978;61:1161—1167.

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12. Schwaiger M, Schelbert HR. Ellison D, et al. Sustainedregional abnormalities in cardiac metabolism aftertransient ischemia in the chronic dog model. J AmCoilCardiol1985;6:336—347.

13. Sochor H, Schwaiger M, Schelbert HR, et al. Relationship between 11-201, Tc-99m (Sn) pyrophosphate andF-i8-2-deoxyglucose uptake in ischemically injureddogmyocardium.Am HeartJ 1987;114:1066—1077.

14. Sokoloff L, Reivich M, Kennedy C, et al. The (‘4C)deoxyglucose method for the measurement of localcerebral glucose utilization: theory, procedure, andnormal values in the conscious and anesthetized albino rat. JNeurochem 1977; 28:897—916.

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16. Krivokapich J, Huang S-C, 5dm CE, et al. Fluorodeoxyglucose rate constants, lumped constant, andglucose metabolic rate in rabbit heart. Am J Physiol1987; 252:H777—H787.

17. Thakur ML, Segal AW, Welch LL, et al. Indium-i 11-labeled cellular blood components: mechanism oflabeling and intracellular localization in human neutrophils. JNuclMed 1977; 18:1022—1026.

18. Peters AM, Saverymuttu SH, Reavy HJ, et al. Imagingof inflammation with indium-i 11 tropolonate labeledleukocytes. JNuclMed 1983; 24:39—44.

19. Weiss ES, Ahmed SA, Thakur ML, et al. Imaging ofthe inflammatory response in ischemic canine myocardium with ‘‘‘Indium-labeledleukocytes. Am J Cardiol1977;40:195—199.

20. Thakur ML, Gottschalk A, Zaret BL. Imaging experimental myocardial infarction with indium-i I 1-labeled autologous leukocytes: effectsof infarct age andredidual regional myocardial blood flow. Circulation1979; 60:297—305.

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1832 Wijns,Melin,Lenersetal The Journalof NuclearMedicine

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1988;29:1826-1832.J Nucl Med.   Christian Michel, Anne Bol, Annie Robert, Hubert Pouleur, André Charlier and Christian BeckersWilliam Wijns, Jacques A. Melin, Norbert Leners, Augustin Ferrant, André Keyeux, Jacques Rahier, Michel Cogneau,  UptakeMyocardium: Relation with Tissue Viability Assessed by Fluorine- 18-2-Deoxy glucose Accumulation of Polymorphonuclears Leukocytes in Reperfused Ischemic Canine

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