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Comparison of euro collins®- and low potassium dextran (perfadex®) -solution in clinical lung preservation
The Journal of Heart and Lung Transplantation January 1999
40 Abstracts
14
Baekgroud: Myocardial ptewvatiott temaim a majcz pl in clinical bat uamph~tiat.hprdfoluthcpntceliond~mdcpadcntand~ vasomotorfunctiooasapedie(orolhrnsplmt~~aseascandloog-tam survival is d msja impmbnm. We inwstigakd the effecta d C&a schticm, a sndcndnheliem MW hatt pmsctvstial sdntion on left VcnaicuLr pctfawnce &pcmicataadindcpen&nt vrsomdafimctiatdca2hdgbbal isckmia in is&ted railKalts.Methodsr Isdatalkattswe$emamtsdata~Appamtusand pcdllsed with muwkd KJcbs-melt bdfes. Heat rate (Em), ldt vellhicnlst tkvdqml pgauc (LVP) ad eddiaaDdie pesam (LVEDP), max. positive (+dP/dt) and mgstive dP/dt (dmt). (-Iammy llow (CF) and c4umlmy flow msetvc (CFR) to .$z&wilmt dlzpdat (&adyhinin @sdy, l@vhink5 min) aId hdqndmt (Admositte (Ade; 10 pM/tnid4 mbl) stimdi mn dttambd bcfae and dtu ischemia. Cdsior adutiott (20 ml/kg) WOB perfused at a constant pmssurc d 50 mmHgviatkaorticmo~Resaltm After2hdiechcmiafollowcdbylhof trqerfusia~ ctmqmble m-snlts (ANOVA) between pm- nod postiscbcmic hcmodymnics wa obeewed. HR: prr: 297i17, post: 272*18; LVDP: prc: 82.1i10.5, pear 71.116.0; LVEJX pre: 19.7i5.6, pee’: 18.2i5.8; +dP/dtz pre: 207W61.post: 197fk218; dPl& -1474i.130. post: 144&171. p=n.8. In addition. pm sml postisctic CF (mUmin) did not dilTer attmng gmttps @rc: 135.k 1.5, post: 13.li.8. p=.74). In caatxmt, p~iscbmttic embthe&tma depaaclll ami ittdepcmknt CFR (% increase d CF versus bmdine in rcqcmsc to Bmdy or Adc) WBB sigaitidy impid (CFR-Bmdy: pe vs bweliw 105% p<.OOOl; post w toselinc: 39.5%. 1~07; pr YS post: p=.oo8; CFR-Adc pte “8 baseline: 122%, ~.0001; poet vs baseline: 42.2%. pz.05: prc “8 post: p=.ODZ). Concladoa: The-w data indicate that c%im solntim povidc.9 calls~al wim legad to left hemdym&s; lhnmver, EQoDlIy vasaaotor ftmctkm to endowimn ~I$ZZ snd idqxded sdmdi is signitiwdy impaired. This may lx an impmtantearlymedmniamcc&htlngtotkdcwlopmatdtmns@tcaraary sttety discssc. the majot Iiitiq factor for long tam sorvival &et catdiac kansplantsiion.
15
17
MYOCARDIAL HLA AND ICAMSTAINING PATTERN IN BIOPSIES FROM TRANSPLANTED HEARTS. THREE YEARS FOLLOW-UP R. Wojoicz, M. Zembala, M. Zakliczyliski, J. Nowski, M.M. Rozek. Silesiae Centre of Heart Disease, Zabrze, Poland and HSC at San Antonio, TX, USA The up-regulation of HLA-I (ABC), HLA-II (DR), and intercellular adhesion molecule-l (ICAM-I) in the myocardium after heart transplantation (OHT) has been implicated in organ rejection. The aim of this study was to investigate changes in HLA and ICAMexpression during three years after OHT. Accordingly, we studied multiple endomyocardial biopsy specimens collected t?om 11 randomly selected patients (pts) (all males, mean age 46.5 i- 11.6 years). OHT was performed to treat ischemic cardiomyopathy in 6 pts, (55%) and idiopathic dilated cardiomyopathy in 5 pts (45%). All patients were on a triple immuwsoppressive regimen (cyclosporine, azadtioprine, and prednisolone) up to one year. Biopsy specimens were collected at 1 week, 1 and 3 months, and 1, 2 and 3 years post OHT and evahmted for routine rejection signs. The presence of HLA and ICAM-I were assessed by immtmohistochemistry. The expression of tested antigens was studied on frozen sections using monoclonal antibodies and LSAB@+iKit/New Fuchsin Chromogen (all from DAK0 A/S). Immtmoreactivity (IR) was measured using a semiquantitative scoring system: 0 - no staining; 1 - focally distributed interstitial cells staining; 2 - diffise interstitial staining; 3 - diffuse interstitial and concomitant focal myocyte staining; 4 - diffuse interstitial and myocyte staining. IR > 3 was considered positive for all tested molecules. IR for all studied molecules was negative in 9 pts at 1 week and 1 month post OHT. In 2 pts, IR that was positive for at least one molecule had correlated to an acute rejection episode. At 3 months, IR was positive in 2 pts without histological signs of rejection. One year post OHT all studied molecules had increased expression in 10 pts (91%) and remained upregulated on biopsy specimens collected after 2 and 3 years post surgery. When comparing 1 year verses 7 days, the results were significantly different: HLA-DR (p=O.O04), HLA-ABC (p=O.OOl) and ICAM- (p=O.Ol). There was no correlation between IR and histological diagnosis of rejection. We also observed striking differences in staining pattern between early and late biopsies. The focally distributed and interstitial staining in the early period is replaced with the diffise staining, including sarcolemmal myocyte staining with no concomitant histologic signs of myocyte injury at the late (Z 1 year) period post OHT. This diffused, upregulated expression after 1, 2 and 3 years post OHT supports the hypothesis that the myocardium undergoes “immunological accommodation”.
16
TEMPORAL PATTERN OFT LYMPHOCYTE AND MACROPHACE INFILTRATION IN A NONIMMUNOSUPPRESSED MODEL OF CAV M.P. Fischbein, J.J. Yen, H. Laks, M.C. Fishbein, M.Espejo, K. Ebrahimi, B. Bonavida, A. Ardehali, University of California, Los Angeles Interpreting experimental studies of cardiac allografl vasculopathy (CAV) is confounded by the use of diverse immunosuppressive regimens. We have previously described a murine model of CAV without immunosuppression. In this study, we sought to determine the temporal pattern of cellular infiltrate and intimal proliferation. Methods: Hearts from Bl0.A mice were transplanted into BIO.BR recipients (MHC 1 incompatible). Allografts were harvested at days 0, I, 7, 14, 30, and 50 (n=6 each group). Isografts were analyzed at the same time points. CD4, CD8, macrophages, ICAM, VCAM, smooth muscle actin, MHC I, and MHC II expression
COMPARISON OF EURO COLLINS=AND LOW POTASSIUM DEXTRAN (PERFADE@) -SOLUTION IN CLINICAL LUNG PRESERVATION C. Miillert, H. Hoffmann’, H. Reichenspurner*, F. Kurz, H. Fiirst’ and Munich Lung Transplant Group Department of Surgery’ and Department of Cardiac Surgery* Ludwig Maximilians University, MarchioninistraBe 15, 81377 Munich, FRG In experimental lung transplantation, the superiority of low potassium dextrane (LPD, Perfadex”) over Euro Collins@ solution (EC) was demonstrated. A clinical study was performed to evaluate the effect of LPD in human lung preservation. Method: Patients undergoing single- (SLTx) or double- (DLTx) lung transplantation were investigated. Donor lungs were flushed with EC (n=48) or LPD (n=30, both 50 ml/kg), insufflated with 02 (FiO,=l,O) and stored in a hypothermic state. Duration of ischemia, ventilation and stay on ICU were analized. lschemia - reperfusion injury (If+) was classified in grade 0 (no IRI), grade 1 (mild interstitial fluid accumulation), grade 2 (ventilation > 3 days due to IRI). grade 3 (ventilation with NO, Prostactlandine: ECLA) and arade 4 (death due to IRI).
Conclusion: Despite of a longer ischemic time, there was no IRI grade 4 and fewer grade 3 in the LPD group leading to a shorter ICU stay. Even in clinical lung preservation, the use of LPD (Perfade@) appears to be superior to Euro Collins’“’ solution, leading to a reduction of incidence and severity of IRI
16
involved in clfam&g the nHgnitndc of tbe immune rcaponw. we have employed the Lewis RT’+ Fisher 344 RT’ Hehotopic Gtdiac Allogmft Modd to $ttdy apopt&sinhaYtmjection. EothfmzeslMdpamflin~bsddedhistologi~spctrons horn cadac dlogmftn of animals m&iccd et 10,23,38,45 and 46 days (d) pattrrmsplaa~on (FT) reepective4y, wem hdied by the TUNEL-immmqmmxidase asssy and immmohistochsmistry using antibodies to FM tigand (Fuda) and B&2. Morphdogically, all cardiac dlogmfta exhibited rejection. 10-d PT cardiac sllogdts shmved mild to nwdemte rejsctioo cbamctaizod by local monotmclear irdlammstory cdl intiltmtes (MNC) without significsnt nccrotiring chqeu in the myocardinm. By contrast, 234, 38-d, 45-d and 464 PT cudiac alfogmRs exhibited, hweaaingly, duogee of severe njectiott typified by brisk, multifocal, and partly contlnmt MM!+ necrosis of cardiac myocytes, sod &lamma&ry nod fibmoblitemtivewcdar pathologies. Overlappii perivtinr (PV)and tmttsmtttal vpscditissodothe.Ulitis (TVE) nagioceotric patterns constitute P wrdimtom culminating in P fibmoblitemtive srtexiopathy. TUNEL locpliptian wm detected predomioantly in mhpopolatiom of pn8iocezlttic and iotemtitial MNC (lymphocytes and mooocytes). A lesser number of TlJNBL-pwitivc MNC wp8 dekcted in areas of eudothelilitis. Rare codothdial-like cells in TVE, and rate fibmblast-like roeaedsymsl cells in atw of iote-mtitial fibrosis exhibited TUNEL shining. There wss s paucity of TUNBL localimtion in cardiac myocytcs. Sobpopohtiotw of chiefly PV MNCs io cardiac allograh horn 23-6 PT omwrde, exhibited Fm L sod Bcl-2 immtmoreactivities. Our tinding indicate that qoptoeis is pmdomhotly a feature of the cells of the immtme system in cardii allogmfh rejechn, whereas necrosis wss the main prhdogic comIste of the. attendat myowdial iojwy. Supported in put by PO1 Al40161 and T32 AI07101 from NIH.
Elfocts d Celsiat Presenation Solution on Myocardid Perfmmtmce andCoronarY vasomotor Fmwiat afrcr 2 h of GLobal Ischemia io Isolated Rat Hearts S.M. WILDHlRT, N. CONRAD. S. KUEHh’AF’FEL, H. AKDEMIR, C. SCHUIZE, D.H. BOEHM. B. RJXHART Ikpmfmd d Cardiac Surecry, Ludwig-Maximiliens University. Mm&h, Germany
APOPTOSIS
IN RAT
CARDIAC
ASOCIMEDMTI’EcELLsOFTBE
ALLooRAFr
IMMUNE
Is
PREDoMl.NANrLY
RESPONSE.
A. A@n’, C.D. Kats&c@, L.I. Gkkas’, B. Goldmu?, E.L. Oleazak’, C.D. Pht8owas’, Dept. Microbid. & I mmmd.‘, Dept. P&ml.‘, Fds ht. Cancer Rea. Br Mol. Bid.‘, Temple Univ. Sch. of Med., Dept. Pediattid, St. Christopher’s Hosp. Child, MCP/Hhwmam Sch. Med., Phihddphis, PA. infiltmtelardiacldlolpftsMdplayno T~ad-Ytd~ importantmleiobeut+ction. htbiscontext,qoptosisisoneoftbeme&nisms
40 Abstracts
14
Baekgroud: Myocardial ptewvatiott temaim a majcz pl in clinical bat uamph~tiat.hprdfoluthcpntceliond~mdcpadcntand~ vasomotorfunctiooasapedie(orolhrnsplmt~~aseascandloog-tam survival is d msja impmbnm. We inwstigakd the effecta d C&a schticm, a sndcndnheliem MW hatt pmsctvstial sdntion on left VcnaicuLr pctfawnce &pcmicataadindcpen&nt vrsomdafimctiatdca2hdgbbal isckmia in is&ted railKalts.Methodsr Isdatalkattswe$emamtsdata~Appamtusand pcdllsed with muwkd KJcbs-melt bdfes. Heat rate (Em), ldt vellhicnlst tkvdqml pgauc (LVP) ad eddiaaDdie pesam (LVEDP), max. positive (+dP/dt) and mgstive dP/dt (dmt). (-Iammy llow (CF) and c4umlmy flow msetvc (CFR) to .$z&wilmt dlzpdat (&adyhinin @sdy, l@vhink5 min) aId hdqndmt (Admositte (Ade; 10 pM/tnid4 mbl) stimdi mn dttambd bcfae and dtu ischemia. Cdsior adutiott (20 ml/kg) WOB perfused at a constant pmssurc d 50 mmHgviatkaorticmo~Resaltm After2hdiechcmiafollowcdbylhof trqerfusia~ ctmqmble m-snlts (ANOVA) between pm- nod postiscbcmic hcmodymnics wa obeewed. HR: prr: 297i17, post: 272*18; LVDP: prc: 82.1i10.5, pear 71.116.0; LVEJX pre: 19.7i5.6, pee’: 18.2i5.8; +dP/dtz pre: 207W61.post: 197fk218; dPl& -1474i.130. post: 144&171. p=n.8. In addition. pm sml postisctic CF (mUmin) did not dilTer attmng gmttps @rc: 135.k 1.5, post: 13.li.8. p=.74). In caatxmt, p~iscbmttic embthe&tma depaaclll ami ittdepcmknt CFR (% increase d CF versus bmdine in rcqcmsc to Bmdy or Adc) WBB sigaitidy impid (CFR-Bmdy: pe vs bweliw 105% p<.OOOl; post w toselinc: 39.5%. 1~07; pr YS post: p=.oo8; CFR-Adc pte “8 baseline: 122%, ~.0001; poet vs baseline: 42.2%. pz.05: prc “8 post: p=.ODZ). Concladoa: The-w data indicate that c%im solntim povidc.9 calls~al wim legad to left hemdym&s; lhnmver, EQoDlIy vasaaotor ftmctkm to endowimn ~I$ZZ snd idqxded sdmdi is signitiwdy impaired. This may lx an impmtantearlymedmniamcc&htlngtotkdcwlopmatdtmns@tcaraary sttety discssc. the majot Iiitiq factor for long tam sorvival &et catdiac kansplantsiion.
15
17
MYOCARDIAL HLA AND ICAMSTAINING PATTERN IN BIOPSIES FROM TRANSPLANTED HEARTS. THREE YEARS FOLLOW-UP R. Wojoicz, M. Zembala, M. Zakliczyliski, J. Nowski, M.M. Rozek. Silesiae Centre of Heart Disease, Zabrze, Poland and HSC at San Antonio, TX, USA The up-regulation of HLA-I (ABC), HLA-II (DR), and intercellular adhesion molecule-l (ICAM-I) in the myocardium after heart transplantation (OHT) has been implicated in organ rejection. The aim of this study was to investigate changes in HLA and ICAMexpression during three years after OHT. Accordingly, we studied multiple endomyocardial biopsy specimens collected t?om 11 randomly selected patients (pts) (all males, mean age 46.5 i- 11.6 years). OHT was performed to treat ischemic cardiomyopathy in 6 pts, (55%) and idiopathic dilated cardiomyopathy in 5 pts (45%). All patients were on a triple immuwsoppressive regimen (cyclosporine, azadtioprine, and prednisolone) up to one year. Biopsy specimens were collected at 1 week, 1 and 3 months, and 1, 2 and 3 years post OHT and evahmted for routine rejection signs. The presence of HLA and ICAM-I were assessed by immtmohistochemistry. The expression of tested antigens was studied on frozen sections using monoclonal antibodies and LSAB@+iKit/New Fuchsin Chromogen (all from DAK0 A/S). Immtmoreactivity (IR) was measured using a semiquantitative scoring system: 0 - no staining; 1 - focally distributed interstitial cells staining; 2 - diffise interstitial staining; 3 - diffuse interstitial and concomitant focal myocyte staining; 4 - diffuse interstitial and myocyte staining. IR > 3 was considered positive for all tested molecules. IR for all studied molecules was negative in 9 pts at 1 week and 1 month post OHT. In 2 pts, IR that was positive for at least one molecule had correlated to an acute rejection episode. At 3 months, IR was positive in 2 pts without histological signs of rejection. One year post OHT all studied molecules had increased expression in 10 pts (91%) and remained upregulated on biopsy specimens collected after 2 and 3 years post surgery. When comparing 1 year verses 7 days, the results were significantly different: HLA-DR (p=O.O04), HLA-ABC (p=O.OOl) and ICAM- (p=O.Ol). There was no correlation between IR and histological diagnosis of rejection. We also observed striking differences in staining pattern between early and late biopsies. The focally distributed and interstitial staining in the early period is replaced with the diffise staining, including sarcolemmal myocyte staining with no concomitant histologic signs of myocyte injury at the late (Z 1 year) period post OHT. This diffused, upregulated expression after 1, 2 and 3 years post OHT supports the hypothesis that the myocardium undergoes “immunological accommodation”.
16
TEMPORAL PATTERN OFT LYMPHOCYTE AND MACROPHACE INFILTRATION IN A NONIMMUNOSUPPRESSED MODEL OF CAV M.P. Fischbein, J.J. Yen, H. Laks, M.C. Fishbein, M.Espejo, K. Ebrahimi, B. Bonavida, A. Ardehali, University of California, Los Angeles Interpreting experimental studies of cardiac allografl vasculopathy (CAV) is confounded by the use of diverse immunosuppressive regimens. We have previously described a murine model of CAV without immunosuppression. In this study, we sought to determine the temporal pattern of cellular infiltrate and intimal proliferation. Methods: Hearts from Bl0.A mice were transplanted into BIO.BR recipients (MHC 1 incompatible). Allografts were harvested at days 0, I, 7, 14, 30, and 50 (n=6 each group). Isografts were analyzed at the same time points. CD4, CD8, macrophages, ICAM, VCAM, smooth muscle actin, MHC I, and MHC II expression
COMPARISON OF EURO COLLINS=AND LOW POTASSIUM DEXTRAN (PERFADE@) -SOLUTION IN CLINICAL LUNG PRESERVATION C. Miillert, H. Hoffmann’, H. Reichenspurner*, F. Kurz, H. Fiirst’ and Munich Lung Transplant Group Department of Surgery’ and Department of Cardiac Surgery* Ludwig Maximilians University, MarchioninistraBe 15, 81377 Munich, FRG In experimental lung transplantation, the superiority of low potassium dextrane (LPD, Perfadex”) over Euro Collins@ solution (EC) was demonstrated. A clinical study was performed to evaluate the effect of LPD in human lung preservation. Method: Patients undergoing single- (SLTx) or double- (DLTx) lung transplantation were investigated. Donor lungs were flushed with EC (n=48) or LPD (n=30, both 50 ml/kg), insufflated with 02 (FiO,=l,O) and stored in a hypothermic state. Duration of ischemia, ventilation and stay on ICU were analized. lschemia - reperfusion injury (If+) was classified in grade 0 (no IRI), grade 1 (mild interstitial fluid accumulation), grade 2 (ventilation > 3 days due to IRI). grade 3 (ventilation with NO, Prostactlandine: ECLA) and arade 4 (death due to IRI).
Conclusion: Despite of a longer ischemic time, there was no IRI grade 4 and fewer grade 3 in the LPD group leading to a shorter ICU stay. Even in clinical lung preservation, the use of LPD (Perfade@) appears to be superior to Euro Collins’“’ solution, leading to a reduction of incidence and severity of IRI
16
involved in clfam&g the nHgnitndc of tbe immune rcaponw. we have employed the Lewis RT’+ Fisher 344 RT’ Hehotopic Gtdiac Allogmft Modd to $ttdy apopt&sinhaYtmjection. EothfmzeslMdpamflin~bsddedhistologi~spctrons horn cadac dlogmftn of animals m&iccd et 10,23,38,45 and 46 days (d) pattrrmsplaa~on (FT) reepective4y, wem hdied by the TUNEL-immmqmmxidase asssy and immmohistochsmistry using antibodies to FM tigand (Fuda) and B&2. Morphdogically, all cardiac dlogmfta exhibited rejection. 10-d PT cardiac sllogdts shmved mild to nwdemte rejsctioo cbamctaizod by local monotmclear irdlammstory cdl intiltmtes (MNC) without significsnt nccrotiring chqeu in the myocardinm. By contrast, 234, 38-d, 45-d and 464 PT cudiac alfogmRs exhibited, hweaaingly, duogee of severe njectiott typified by brisk, multifocal, and partly contlnmt MM!+ necrosis of cardiac myocytes, sod &lamma&ry nod fibmoblitemtivewcdar pathologies. Overlappii perivtinr (PV)and tmttsmtttal vpscditissodothe.Ulitis (TVE) nagioceotric patterns constitute P wrdimtom culminating in P fibmoblitemtive srtexiopathy. TUNEL locpliptian wm detected predomioantly in mhpopolatiom of pn8iocezlttic and iotemtitial MNC (lymphocytes and mooocytes). A lesser number of TlJNBL-pwitivc MNC wp8 dekcted in areas of eudothelilitis. Rare codothdial-like cells in TVE, and rate fibmblast-like roeaedsymsl cells in atw of iote-mtitial fibrosis exhibited TUNEL shining. There wss s paucity of TUNBL localimtion in cardiac myocytcs. Sobpopohtiotw of chiefly PV MNCs io cardiac allograh horn 23-6 PT omwrde, exhibited Fm L sod Bcl-2 immtmoreactivities. Our tinding indicate that qoptoeis is pmdomhotly a feature of the cells of the immtme system in cardii allogmfh rejechn, whereas necrosis wss the main prhdogic comIste of the. attendat myowdial iojwy. Supported in put by PO1 Al40161 and T32 AI07101 from NIH.
Elfocts d Celsiat Presenation Solution on Myocardid Perfmmtmce andCoronarY vasomotor Fmwiat afrcr 2 h of GLobal Ischemia io Isolated Rat Hearts S.M. WILDHlRT, N. CONRAD. S. KUEHh’AF’FEL, H. AKDEMIR, C. SCHUIZE, D.H. BOEHM. B. RJXHART Ikpmfmd d Cardiac Surecry, Ludwig-Maximiliens University. Mm&h, Germany
APOPTOSIS
IN RAT
CARDIAC
ASOCIMEDMTI’EcELLsOFTBE
ALLooRAFr
IMMUNE
Is
PREDoMl.NANrLY
RESPONSE.
A. A@n’, C.D. Kats&c@, L.I. Gkkas’, B. Goldmu?, E.L. Oleazak’, C.D. Pht8owas’, Dept. Microbid. & I mmmd.‘, Dept. P&ml.‘, Fds ht. Cancer Rea. Br Mol. Bid.‘, Temple Univ. Sch. of Med., Dept. Pediattid, St. Christopher’s Hosp. Child, MCP/Hhwmam Sch. Med., Phihddphis, PA. infiltmtelardiacldlolpftsMdplayno T~ad-Ytd~ importantmleiobeut+ction. htbiscontext,qoptosisisoneoftbeme&nisms