Relationship between angiotensinogen, a l-protease inhibitor elastase complex, antithrombin III and C-reactive protein in septic ARDS U.l-lilgenfeldt1, W. KeUenllann2, G . Kienapfel !, and M. Jochum3
I Dcpartmem of Pharmacology, University of Hcidelbcrg, 1 Dcpartment of Anesthesjology, CHnic Großhadcm and 1 Dcpartmcnl of CJinical Chemislry and Biochemistry. City Clinic, University of Munieh, FRG
Rccci\'cd: May 23. 1989/Accepted in revised form: September 12, 1989
Summary. The time-course of plasma angiotensinogen (Ao), elastase-Cll-prolease inhibitor complex (Ew\PI), antithrombin 1II (AT lli) aod C-reactive protein (eR!» have been invest igated ofsix patients suffering from adult respiralory distrcss syndrome (AROS).
Thc total plasma Ao level (activc and inactive Ao) vancd in individuals but was increascd up to five-fold. An incrcasing amount of inactive Ao is found. From the beginning oe their stay in the intensive care unil up to five days haU of the patients displayed a positive eorrelation bctween the pla~ma eRP and Ao level. Thc CRP and Ao values were either nol or were negativeJy correlated with the AT 111 val ues.ln contrast plasma Ao and AT Iß levels in all patients were positively correlated during a particular period in the subsequent phase of the disease, where there was no or a negative corrclation with CRP. The two acule phase reaelants CRP and ELaIPI were only correlaled in two palients at the beginning ofthc disease.
The markcdly increased plasma level allhe beginning of the inflam matory diseasc indicates that Ao is an acutc phase reaclanl, and this is supported by the parallel changes in plasma CRP and Ao levels during the early days of ARDS. The relationship between the plasma levels of Ao and AT 1lI for more than fourteen days suggcsts similar regulation of these members oi thc serpin family after termination oitbe acutc-phasc.
Key words: angiolensinogen, antithrombin, acute-phase proteins, septie ARDS; elaslase-al-protease inhibitor eomplex, C-rcacüve-protein, renin-angiotensin system, adult respiratory distrcss syndrome, sepsis
Angiote nsinogen (Ao) is thc only known precursor of the vasoactive pepl ide hormone angiolcnsin. This plasma glycoprotein is synthesizcd and secrelcd by the liver. Various hormones are known to be involved in ils synthesis and secretion, inc1uding angiotensin 11 (Khayyal et al., 1973), oestrogens and glucocorticoids (Crane el al., 1971), and thyroid hormones (Dzau and Hcrrmann, 1982). Subseq uent to publication of the cDNA sequence of Ao by
Ohkubo and coworkers (1983) , a sequenccrelationship to the serpin (serine protease inhibitors) family, antithrombin 111 (AT 1lI), ovalbumin and cxpecially 10 arprotease inhibitor (alPt) was detccled by Dool ittle (1983). This was supported by the finding oi Tanaka cl al. (1984), showing a common Slructural organizalion ofthe Ao and a lPI genes.alPl interacts with lysosomal enzymes, inc1uding kallikrein, various cathepsins and especially elaSlase to form the elastase-al-protease inhibitor compIex (ELa1PI). A III is tbc most important inhibitor oflheclolting system (Travis and Salvescn, 1983). AT III and al PI are acute-phase reactants in various an imals (Koj and Regoecz.i, 1978; Kushner, 1982), whereas in humans A fII is not (Martfnez-Brot6ns CI a1., 1987) . C-reactive protein (CRP) is a further acute-phase reactant, whieh does not belang to the serpin family. The physiological role ofCRP is not yet comletely undcrstood (McCarty, 1982). Recenl data have shown that CRP-peptides are poten t imm unomodulators (Robey ct al., ]987). Wilhin hours fo llowi ng acule inflammation, thc syn thesis of these protcins is increased in the Iiver and their plasma concentration i5 elevatcd by as much as 2000-fold (Koj and Gordon, 1983; Morley und Kushncr, 1982).11 has recently been shown that administration of lipopolysaccharides (LPS) 10 rats resulls ina marked increase in a lP] and Ao mRNA in the liver (Kageyamaet a1. , 1983) and an increase in plasma Ao levels (Hilgenfeldt, 1988). These find ingsindicate that Ao is a member of the acute-phase reaclant dass in rats. To verify its role in inflammatory diseases in men, the plasma levels of Ao have been compared over a 4 weck period with those of EuIPI, AT III and CRP in patients suffering from septic ARDS (Madig, 1986).
Materials ::IOd methods
Patients
Plasma sampies from six patients (3 male, 3 female) with a mean age of 33 (18.5, SO) Y and a mean weight of67.7 kg, admitted 10 the Intensive Care Uni\, Klinikum Großhadern, University of Munich
Ao! (omol Alfml)
10
0.1 ..•.•
0.1
.. ' .... ....
.... .. '
-/ ... ~" . . '
r=0.869
Aod (nmol!ml)
,,' ..... /"
r=O.935
10
~·ig.l. Plasma angiotensmogen in eontrol group obtained by dircet (Aod) and indireet (Aoi) radioimmunoassay (centre group), n = 24. Mean (SO) 0.88 (0.24) nmol Aolml. Plasma angiotensinogen in pregmIßt women (right group), 11:: 12,6.2 (1.3) nmol Aofml. The solid liDe is the calculated corrclation betwccn the assays and the dotted line shows the theoretical l: I corrclation
were invcstigated. Qinical data are given in Table 1. Only patients kept in the Intensive Gare Unit for more than 10 days were eligible for thc study. These critical1y iU patients all suffered from bacterial sepsis and the adult respiratory distress syndrome (ARDS) (Modig, 1986).
All patients rcceived ant ibiotics, low dose heparin , diazeparn, morphine and pancuronium bromide. In addition, furosemide was administered ir urine flow was reduced. Epincphrine and dopamine were administered to five of the six patients (exocpt PI0) to maintain blood press ure and kidney function.
Crileria for exclusioll. Patients werc excluded in cases of negative blood cu[ture or no focus of severc infection. Patients wilh liver failurc cvideneed by history and laboratory tests, with ncoplasticdisease, with any chron ie illncss, or who rcceived glucocorticoids or oral contraceptives were also exc1uded.
Preparation of plasma. Up to four blood sam pies per day were obtained [rom indwelling venous catheters. Blood wa~ eollected in 5 ml pla,>tic tubes containing O.S ml trisodium citrate 0.13 M. Plasma was prcpared by eentrifugation ofcitrated blood at 1000 g for 10 min at room temperature and fr07,.en at - 70'C in plustic tubcs.
Assay of angiolensinogen. Ao was completc1y purified from plasma obtained from the Department of Gynaecology, University of Heidelberg by a five step procedure. Purity was verified by amino aeid analysis, end group sequence analysis and oomplete liberation of N-teTminal angiotensin I by renin. The protein had a specificactivity of 2J.Jl1g ANG I·mg - t protein. Asuming a moleeular wcight of 6O,0IXl Da for human angiotensinogen, the preparation has a punty of 98% (Kienapfel, 1987). Antibodies against human Aowere raised in rabbits by thc method of Vaitukaitis et al. (1971). Plasma Ao ooncentrations were determincd bya direct radioimmunoassay (RIA) perfonned according to lhe method described for rat Ao (Hilgenfeldt and Schott, 1987). Antiserum was uscd at 1 :2501Xl. The sensitivity of the assay was 1 fmol Ao. In addition, an indirect assay following enzymatic cleavage o[ Ang l with an exces.s of renin (Hilgenfc1dt and Haekcnthal, 1979), was also used. Angl was measurcd by R1A (Menard and Calt, 1972). Ang l antiserum was raiscd in rabbiL~ using glutaraldehydc-linked angioteusin I. The serum was analY-Led for cross-reaetivity with Ang ]\. The antibody titre for AngI and AngIl differed by S.Hr.
Assay 0/ elastase-a)-proleinase inhibitor complex (EICl.[ Pl). ELatPI complcx was measured using a commercial, solid phase, enzymelinked, sandwich immunoassay (E. Merck AG, Darmstadt, FRG), as described by Joehum ct al. (1983). The range was from 20 to 150 Ilg (l)P I-complex per Htre.
Assay of plasma anlithrombin (ATlU). Plasma AT UI concentration was determined wüh a commereial thrombin inhibition assay (S-2238, Deutsche Kabi-Vitrum, Munich) using p-nitroaniline as the cromogenic product.
Assay ofC-reaelive pro/ein (CRP) . The conceulration ofC-rcactive protein was mcasured by radial immunodiffusion using a commereial test system (Behring Werke AG, Marburg, FRG). Thc coeffieient of correlation was ca1culatcd by comparing different parameters of evcry plasma sampie of eaeh patient using thc computer program Microsoft Chan .
Rcsults
Contro! plasma Ao levels
Thc values obtained by the direct and indirect assays for Ao were compared by measuring tbe angiotensinogen conte nt in plasma irom 25 heall hy volunteers. The plasma Ao levels were 0.876 (0.278) nmol Ao·ml - J and 0.874 (0.237) nmol Ang I .m1 - 1 (mean (SO). This indicates that in hcalthy man the plasma Ao f1 uctuates within a narrow range. In addition plasma fTom 10 healthy pregnant women, with the pregnancy dependent elevation of Ao was also tested. The mean plasma Ao was6.19(1.25) nmol Ao·ml - 1 and 6.07 (1.6) nmol AngI .ml - 1• As shown in Fig. 1, the correlation coefficicnt betwecn thc direcl and indirect assays for Ao in both groups was dose to].
Patients
Ao, ELcxIPI , AT u r and CRP were analyzed in 209 different plasma sampies from 6 patients. As shown in Fig.2, plasma Ao followed an unique time course in each pa-
Tahle L a) Traumatic event (days prior admission; Day 0 = bcginrung of the sludy, Day of admission to the intensive care uni!; m ", male.f = female) b) Clinicaldiagnosis(n =6)
P9 a) 10 (f), Dear drowning b) Severe ARDS two days prior admission, Legionella
pneumophilia , sepsis syndrome on Days 8-IOof thc study with aeutc renal insuf[iciency, rhabdomyolysis and death on Day 27.
P 10 a) S (m) polytrauma. b) Pneumonia, sepsis (Slaph. al/reus, Ps. aerugil1ol"U),
ARDS. On Day 6 aeute renal insu(fieiency. Second infcction on Day 16.
P I7 a) 8h(m)Polytraumaandsevere posttraumatic ARDS.
b) At Day 10 pneumonia, at Day 18-20sepsis and acute renal insufficicncy.
P45 a) 3 (f), sepsis syndrome 6 h prior admission. b) Shock followed by severe ARDS, reeurrent
infeetions (pneumonia, urinary traet infections). On Day 3 aeute renal insufficiency.
P48 a) OCr), hysterectomy. b) postoperative sepsis (Cl. perjringens), severc
consumptive coagulopathy. Day 3 re-[aparotomy
PS3 a) 7 (m), polytrauma. b) Sepsis, scverc ARDS required artificial respi ration
for 24 days. Day 2 acute renal insufficieucy.
P9 '"
30
1
0, 10 15 20 15
j~ 0510152025
"0 .. '" n.,1/11 1
1.5
• 0.'
00 10 15 20 ~S 30 0
0 1 , , 10 12
" 0 .. 1/.1 Pt7 , .. n.al/I'
PS,
: A 1i-~\~ ..... t~u~r l~ o 5 10 15 20 ~ * ~
time (days)
• , 1
PO ElI I?1 U/.l
"'0
'00 '00
100
o !0-"1,--;--",-C';--;;10,OI1;--;I' time (aays)
.. nlHlllll
• , '"
ti g.2. Plasma Aolevels (nmoVml) oblaincd by the dircci (solid symbols) and indircci (open symbols) radioimmunoassay (nmol A IIml) in six patienlswith septic A RDS
ElalPI U/I\
600
500 <00 300 100 100
Os 15 15 0 O~----~----~~-"O o 10 20
PIO
1
OL---------~--__e o 10 20
.. IWIOl/.l
, 1
PI7
10 20 time (days)
30
ELalPI 11/.1
'" '00 <00
0.'
0 0
ElalPI Ujll
.. nllOl/.l
1000 , '" 600
1 <00
100
0
pO,
1 , , 10
PS,
1 6 B 10 12 tlle (days)
ElalPI 11/.1
6000
<000
2000
0
" fLalPJ
11/.1
'" 600
.00 100
l·ig. 3. Plasma Aoand ELa, PI levels in six patients suffering (rom sepsis. The Ao "alues in the dircct radioimmunoassay are consistent with those ofFig.2. (Au solid linc; ELa,PI open symbols)
" " UIII " '" Alill 11I01/al • ßIIOII.I • • '00
'" , ". " 80 " , " ..
• .. • " , 15 " • " " '" ATllI " '" Uill
l\JIo l/., • n.ollll • , 15. 1.' "" ,. "" ,
.. , \...., 90
• • 70 • " " • , • , , " "
" '" " '" Il101/111 UIII MO/Ci I ATlI I • • • 3
~r BO fig.4. Plasma Aoand AT II! levels in , "" six paticntssuffcring from sepsis. Thc ,
lOG Ao valucs in the dircci radio-
" immunoassay are consistcnt with those
" ofFig.2 (Ao solid liDe; AT IIl open
• .. 0 - 60 symbols) • " " 30 • , • , , " " " time l~aysJ Ulle Id~Y$1
" " CRP x, '" CRP n.,l/., Ig/dl fYlo/_ ' ~/dl
" • '" • " , " "
0 0 0 0 , 15 " 0 " '" " ". CRP .. '" CO'
MOI/II '9/dl oaol/il ~/dl , L5 "
'" ,
" .. , • 0 0 • • • " '" 30 0 ,
" " '" CRP .. '" CO'
nllDl/l' IlIJ/dl I\l10/11 IIg/dl
• '" " 3
B . ' ig. S. Plasma Ao and CRP levels in six , palienlssuffcring trom septic A RDS. ,
" TIle Ao valucs in lhe di rect rad io-immunoassay are consistent with those of Fig. 2 (Ao solid line; e RP open
0 0 0 0 symbols) 0 " " 30 0 ,
" t lle (days! U .. (days)
T:.ble 2. Corrclation c()(!fficicnt (r) of plasma Ao levels obtained by direct (d) and indirect (i) RI A and corrcsponding plasma ELa1 PI , ATII I and CRP in six patients suffcring rrom septic ARDS. Thc time·period after admission to thc hospital (days) and as thc isolated valucs (rl) dur· ing this pcriod are given. Values in parenthescs arc the correlatiOD cocfficicnts fo r limitcd time periods
Patient No 9 10
Aod·Aoi 0.51 0.96 (0.814) (0.758) 10.4891 {0.955)
Aod·ELo.1I'1 0.500 0.478 ( - 0.479) ( -0.737) 1- 0.6481 10.4381
Aod·ATIII 0.69 0.038 (0.468) ( -0.476) 10.8841 10.7281
Aod·CRJ> - 0.202 0.494 ( - 0.461) (0.778) 1- 0.2461 10.0451
AT III ·CRP - 0.303 0.488 ( - 0.483) ( -0.402) ! - 0.434) 10.323}
ELa,PJ.CRP - 0. 171 0.351 (0.685) (-0.531) {0.235] 10.405)
time a rte r adm ission (days) O-V 1.3-29 (0-186) (1.3- 5.3) {IO-VI {9 -291
" 37 46 (20) (9) 1171 1311
rient. Thc amount of Ao in all palients was incrcascd at SQmc time during Ihe period of observat ion. Jn addition, the values oblai ned by Ihe dircct and indirect measure· ment show marked di(fere nces. In contrast to the control data given in rig. 1, th e two assay systems did oot show a 1: 1 ratio in plasma from Ihese patients, although the values were st ill correlated (Table 2).
Within a cc rtain pc riod, thc values obtained with tbe direct assay were significant ly higherin P9, P 17, P45 and P 53. This indicales increased formation of Angl, leadiog to an iocreased plasma conccntration of des-AngI-Ao. Stimulation of the components of the renin-angiotensin system, e.g. Ao, ren in, Ang J and Ang n , in some of these patients has reeently been reported (HiJgenfc1dt ct al., 1987). The Ao values obtained by the direct assay werc compared with the Icvc lsofELa1PI, AT IJI and CRP. Plasma Ao and ELa1PI in a ll six patients are shown in Fig.3. A positive correlalion of Ao and ELa1PI was not observed in most patients. Only in P 10 was there nuctuation of the ELa1PI values around the Ao va lues, although they were not positively correlaled; in P 53 r = 0.734 was found wiLhin fi ve days after admission.
Tbc plasma levclsof Aoand AT llJ are shown in Fig.4. A positive corre lat ion of Ao and AT III was evident in plasma from three pa ti ents during Ihe enlire observation period . with r bctwecn 0.69 and 0.71 . The other three pa tients (P 10, P48 and P53) show a good correlation within a linliled period (Tab le 2, dala in parentheses) . alPT and AT III are mcmbers orl he same gelle family and so are rclated to angiotcnsioogcn. However, the acute phase prote in CRP does not belang to this group. The plasma lcvels
17 45 48 53
0.649 0.932 0.709 0.74 (0.967) (0.827) 0.876) 10.5781 ( - 0.5831 10.474)
- 0.580 - 0587 - 0.357 0.489 ( - 0.699) ( - 0.509) (0.734) ( - 0.1351 1- 0.791) 1-0.1231
0.709 0.711 - 0.39 - 0.351 (0.851) ( - 0.679) ( -0.636) 1- 0.0961 {0.8691 10.7151
- 0.294 0.213 0.531 0.579 (0.567) (0.789) (0.784) 10.2371 [0.1071 {0.1441
- 0.417 - 0.190 - 0.793 - 0.825 (0.354) ( - 0.617) ( - 0.706) [ - 0.7111 {- 0.794] {0.0701
0.083 - 0.300 0.233 0.736 ( - 0.233) (- 0.350) (0.634) {- 0.6671 {0.3071 {- 0.4991
0-34.2 0-23.7 0-11 0-13 (0-14) (0- 5) (0- 5) {14-241 15-111 15-131
35 49 16 26 (30) (10) (12) 1191 161 1141
of Ao and CRP arc shown in Fig.5. lt is interesting to note that in P 10, P48 and P S3 the Ao and CRP values were positively corrc1aled att he bcginning of Ihe illness. In the time inlerval in which plasma Ao and CRP were positively correlated, however, Ao aod ATllI showed a negative or no corre lation (see Table 1). A eorrclation between CRP and ELalPI was Icss common. In addition , th ere was 00 or almost a negative correlation between CRP and AT JIJ in plasma [rom all the paticnts.
Discussioll
The mean plasma Ao Icvel in healthy volunteers was 0.9 (0.3) llM. An increase in plasma Ao is observed in pregnancy. in wornen taki ng o ral contraceptives, and in hypertension (Gordon, 1983). The possibilit y that Ao may behave as an aeute-phase protein was first raised by Billg (1972), although he did not rind an increase in plasma Ao Icvels following turpentine injection in the rat. Thc idca that Ao may be an aeute-phase reactant was revivcd after the finding that Ao was gcnetically dosei)' related to (X1PI (Dooliltle, 1983 and Taoaka et al., 1984). The idea was supported by the find ing of as much as a five-fold increase in Ao mRNA in liver (Kageyama et al. 1983) and a threefold increase in plasma Ao levels (Hilgenfeldt, 1988) following lipopolysaccharide administration to rats. A1-thougb these datH are consistent with the hypothesis thai Ao is an acute-phase reaclant in hurnans, caulion is needed in inferring thaI da la Crom one spccics can be applied to another; for example, al Pl is not an acute·phase
reaetant in mice (Baumann el al., 1986), and CRP dem onstrates striking species variations in response 10 tissue injury (Pepys ct al., 1979).
Nielscn and Knudsen (1987) reeently reported a 70% inerease in plasma Ao levels in patients with aeute inflammatory disease, indieating that Ao may be an aeute-phase reaetanl in man. Sioec the regulation of plasma Ao levels is com plex, an increascd level on its own cannot be aceepted as proof of this hypothesis. Thc acute-phase response oceurs in the period immediately following injury and infeetion. Thercfore, thc prcsenl study was designed tofollow the time course of change in Ao levels in patients with bacterial sepsis. Tbe changes were compared with those in known aeute-phase reaetants.
Inereased plasma levels of Ao were found during the first 3 to 5 days of observation in five o[ the six patients. The correlation between Ao and the other aeUle-phase proteins in our study'was based on the direet RlA for Ao. This assay recognizes the total Ao eoneentration and ignores Ao eatabolism by plasma reoin. In addition, both proteins, intaet Ao and des-AngI-Ao, have an almOsl identieal plasma half life in vivo (Hilgenfeldt, 1988) and should be eliminated to a similar exlen t.
The individual course of the disease in eaeh patient makes it difficult 10 find a eommon prineiple. 11 is impossible at present to model such eomplexeurvesof different plasma parameters, so it is only possible to eompare different parameters !rom eaeh plasma sampie and to fit them in a linear eorrelation . A relationship between Ao and alPI was seen in only one patient during five days after admission (P 53; r = 0.73). However, thc assay for Ct.IPI only measured the ELCt.IPI-eomplex and not the total amount of aIPI. Thcrefore , a eorrelation between Ao and alPI would only have been dcteeled if the amount o[ the EwlPI-eomplex and of free a lPI were linked. A far stronger relationship was found betwcen Ao and AT 111, with a signifieant eorrelation during lhe entire observation period in tillee patients (P 9, P 17, P45).ln those patients Da eorrelatiOD of Ao and CRPwas found. The other three patienls, PIO, P48, P53, displayed a good corre· lation between Ao and CRP duriog the initial part of the study (r = 0.78, 0.79, 0.78). OuTing this peTiod bolb parameters were negatively eorrelated wilh ATlll (Ao-ATTII: r~~0.48, ~ 0.68 , and ~ O.64 ; CRP-ATIIl: r = -0040, - 0.62, -0.71). In lhe subsequent period Ao and ATIII displayed a good correlation (r = 0.73, 0.87, 0.72) without being related to CR P.
Tbe good correlation of the plasma levels of Ao and CRP during the first sevcral days of hospitalization suggests that Ao is an aeute-phase reaelan! in man. That this correlation was not fouod in all six patients was mainly due to the fact that data from Ihe initial aeute-phase could only be obtained from one patient (P48). On the other hand, plasma Ao and ATJJI levels showed a farcloscr relationship, although ATIJI is not an aeute-phase protein in man (Martinez-Brot6ns el al., 1987). However this eorrelation is observed during a time period when Ao and eRP are nOI eorrclated. This indicates that in the laner period the aeutc-phase has alrcady been terminated. A eorrelation in the plasma levels of gene tieally related proteins like Ao, alP I and Arm, indicates not only a similar
synlhetie response to stimuli, but in addition it points 10
similar rates of catabolism and elimination, despite tbeir molecular and funetional differenees. There isevidence of inereased Ao eatabolism in acute inflammation, since there is a deerease in the in vivo half-life of o ne form oe Ao in rats following LPS Slimulation. Deereased renal elimination, whieh might be eaused by deereased renal blood flow, has also becn diseussed (Hilgenfeldt, 1988).
AI present a common stimulus for aeute-phase proteins cannot be defined. Yery reeently it has been shown that the synthesis of various aeute-phase proteins is medialed by cytokines such as interleukin-l (iL-l), interIcukin-6 (IL-6) and tumor neerosis [aetor-o. (TNF-a), by a direct action on hepatoeytes (Ramadori et al., 1988). However while the seeretion of a r maeroglobulin is mar· kedly aeeeierated by TL-6, the seere li on ofeystcine proteinase inhibitor and alPI is unaffeeted (Andus Cl al., 1988). On the othcr band, a maerophage derived faetor is able 10 stimulale the synthesis and seeretion both of ATIIT and o.lPI (Hoffmann ct al., 1986). Thc biosynlhcsis ofthe mOSI prominent aeute-phase protein CRP by tbc liver is prc· sumably mediated by inlerleukin-l (Kaplan, 1982; Syin cl al., 1986 and Goldberger et al., 1987). Yery reeently it has been shown that the aeute-phase response of Ao is mediated by interleukin-6 in rat hepatoma eeUs (ltoh et al., 1989), bul this requires the presence of glucocortieoids, suggesting that the laller are essential for Ihe stimulatory effeet.
The role of angiotcnsinogen in inflammation cannot yet be defined. It appeared (rom a previousstudy (Hilgenfeldt el al. , 1987) that thc renin-angiolensinsyslem did not playa role in thc maintenanee of blood pressurc in such patients. Howevcr, angiolensin may be importaot in the immune response during intlammation (Fcrnandez-Cas+ tello el al., 1987).
In eonc1usion, up 10 a five-fold inereasc in plasma Ao levels has been found in paticnts sufferi ng from scptie ARDS. The positive eorrelat ion bctween Ao and CRP during the first days after admittanee to the intensive eare unh indicates that Ao is an acute-phase reactant in man. There was also a good eorrelation between plasma Aoand AT lllieveis during the later stage of disease, suggesting similarities in the synthesis and elimination kinetics of these proteins. Further studies are required 10 define the funetional role of Ao in inflammatory diseascs.
Acknowledgemem. The work was supporlcd by a grant from the Deutsche Forschungsgemeinschaft , Bonn -Bad Godesberg, Fed. Rep. Germany.
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Prof. Dr. U. Hilgenfeldt Pharmakologisches Institut Universität Heidelberg Im Nenenhcimer Feld 366 0 -6900 Heidelberg, FRG