2~I
Clinica Chimica Acta, 58 (1975) 291--294 © Elsevier Scientific Publishing Company, Amsterdam -- Printed in The Netherlands
CCA 6831 SERUM LACTATE DEHYDROGENASE ISOZYMES IN HUMAN LUNG HOMOTRANSPLANTATION
SEVERIN M.G. RINGOIR Transplant Laboratory of the Nephrology Department, Medical Clinic, University Hospital, DePir,telaan, 135, B-9.000 Ghent (Belgium)
(Received August 18, 1974)
Summary Over a 10-month pt'riod, after a fight pulmonary graft in a 23 year old male, 90 serum LDH patterns were obtained; the anaerobic serum LDH-5 and LDH-4 fractions rose in acute rejection periods and during chronic rejection, together with total LDH activity. The determination of serum LDH isoenzymes may be useful to detect rejection in pulmonary transplantation.
Introduction
The assay of serum LDH isoenzymes may be of use in detecting rejection after transplantation of heart [3] or kidneys [4]. We report here the serum total LDH activity and LDH isoenzymes after a human lung homotransplantation with 10 months survival. Methods Total LDH activity was determined by the "backward reaction" method according to Warburg and Christian [8], where the decrease of extinction is analysed spectrophotometrically [2], to allow measurements at 37°C in a barbital buffer. The results are expressed in i~ ternational units per ml. LDH isoenzymes were studied by agar gel elect-ophorcsis on microscope slides according to Wieme [6,7]. Visualisation of the eiectrophoretic pattern was achieved usir.g tetrazolium blue with a modified sandwich technique .and the absorbance c f each fraction was measured using a Vitatron spectrophotometer with attached logarithmic recorder. Case report The patient was a 23 year old male with micronodular silicosis. He under-
292 TABLE I SERUM LDH ACTIVITY
IN
LUNG
HOMOTRANSPLANTATION;
LDH
ISOENZYMES
AND
TOTAL
LDH
Nov
Dec
.Jan
Feb
Mar
Apr
May
Jun
Jul
Aug
Sep
Nor- Confidence mal limits values + a n d - 2 o
21.1 22.1 19.9 18.1 18.5
24.5 29.2 21.8 15.6 6.7
22.6 22.2 20.4 19.5 14.9
20.4 26.1 21.2 20.4 11.6
17.6 24.3 22.6 21.2 14.0
18.5 25.2 22.6 19.8 13.7
18.9 2"i.8 23.5 18.8 10.8
20.3 28.7 21.0 17.0 12.6
16.8 26.7 25.6 19.2 11.4
17.8 21.6 20.6 20.1 19.3
15.9 21.9 21.9 20.6 19.7
43.6 41,1 12.1 3.2 0.0
2.8
1.9
3.4
"7
3.1
2.2
2.4
2.2
3.3
4.6
5.8
0.5
1968
1969
% LDH-1 LDH-2 LDIt-3 LDH-4 LDH-5 Total LDH activity (1.U./mi)
39--54 35----47 9---19 2--5 0--5
(0.3--
0.7)
went a right lung homotransplantation on the 14th of N o v e m b e r 1 9 6 8 and survived until the 10th of September 1969. He had an acute clinical rejection crisis on the 5th postoperative day, another one in January 1 9 6 9 and he finally aied having presented clinical s y m p t o m s o f chronic rejection during the last 2 months of his life [ 1 ]. Results In this 10-month period 90 determinations of serum total LDH activity and LDH isoenzymes were made. The mean results for each m o n t h are presented in Table I. Table II shows the LDH patterns we observed in t w o normal-looking specimens of human lung tissue obtained from t w o p~tients during operations for bronchial carcinoma. It is clear that the a m o u n t of anaerobic M units in lung tissue is much higher than in heart muscle tissue or renal cortex. Discussion The total serum LDH activity was never normal during the entire observa. T A B L E II LDI1 I S O E N Z Y M E
PAT'rERN
IN H U M A N L U N G T I S S U E
L I ) H i s o e n z y m e (%)
LDH-1 LDH-2 LDH-3 LDH-4 LDH-5
Spet:imen I
Specitnen H
6.7 19.6 25.9 24.7 25.1
4.3 20.3 37.9 27.5 9.9
293
tion time. The lowest figures were observed ~n December 1968, April and June 1969; in these periods the patient was doing well clinically. The total serum LDH leve! reached peak values in the immediate postoperative phase, and -with an acute rejection period at the 5th day. High values were seen in January, 1969, a month where an episode of dyspnoea occurred for which the immunosuppressive drug regimen had to be increased. In the last 3 months the total serum LDH activity rose from 3.3 over 4.6 to 5.8 I.U./ml together with a deteriorating clinical condition. The serum LDH-5 and LDH-4 increased during the entire 10-month course after transplantation. This increase is n o t only a percentual one but also an absolute one as the total LDH activity was very high. The aerobic LDH-1 and LDH-2 figures were very low during the same period. As the lung contains more anaerobic M units than aerobic H units P ralease of LDH from a rejected lung might provoke a serum increase of the ana,~robic LDH-5 and LDH-4 fractions. The serum LDH.5 was very high in November 1968, and August and September 1969; these periods correspond, respectively, to an acute clinical rejection episode and to increasing chronic rejection in the months before his death. Low or almost normal serum LDH-5 was present in December 1968, when the patient was doing well clinically. From December to January the serum LDH-5 almost doubled and in January we had to treat the patient for rejection again. Thus the evolution of pulmonary graft rejection in this patient seems to bear a good correlation with the serum LDH.5 values; this becomes evident when we mention the serum LDH-5 increase from 7% on the 4th postoperative day to 17% on the 5th postoperative day, where we had all the clinical symptoms of graft rejection with dyspnoea, fever, fall of pO: and good response on an adequate increase oi immunosuppressive therapy. The persistent high total serum LDH activity and increased anaerobic fractions over the entire follow-up period carried a bad prognosis for the graft and for the patient. We do not think a toxic effect of azathioprinc on the i.;ver could be responsible for the observed serum LDH isoenzyme changes [5] as there was never an increase of serum transaminases, and as there were no hepatitis lesions on autopsy. Conclusion
In this case of pulmonary transplantation in man, the assay of serum LDH isoenzymes and total serum LDH activity r, as been useful in diagnosis of acute rejection and in following chronic rejection, an increa~ of LDH-5 and LDH-4 together with a total LDH activity increase indicating rejection of the graft. Acknowledgement Supported by the Belgian Fonds Voor Wetenschappelijk Geneeskv-dig Onderzoek.
294
References 1 2 3 4 5 6 7 8
F. Derom, F, Barbier ~nd S. Ringoir, J. Thorae. Cardiovasc. Surg., 61 ( 1 9 7 1 ) 835 F. K u b o w i t z and P. Ott, Bioehem. Z.. 314 ( 1 9 4 3 ) 94 J.J. Nora. D.A. Cooley and D.J. Fernbach, New Engl. J. Med., 280 ( 1 9 6 9 ) 1079 S. Ringoir° R.J, Wieme and F. Derom, E x c e r p t a Medica Int. Congress series. No. 179, Proc. E.D.T.A. Dublin. 1968, p. 270 T,E. Starzl. T.L. Marchioro and K.A. Porter, j u : g e r y , ,58 (19C5) 131 R.J. Wieme, Ctin. Chim. Acta, 4 (1959) 317 R.J. Wieme, Aoargel Electrophoresis. T h e o r e t i c a l and Applied, Elsevier, A m s t e r d a m ° 1965 O. W a r b u ~ a id W. Christian, Biochem. Z., 186 (1963) 81
Clinica Chimica Acta, 58 (1975) 291--294 © Elsevier Scientific Publishing Company, Amsterdam -- Printed in The Netherlands
CCA 6831 SERUM LACTATE DEHYDROGENASE ISOZYMES IN HUMAN LUNG HOMOTRANSPLANTATION
SEVERIN M.G. RINGOIR Transplant Laboratory of the Nephrology Department, Medical Clinic, University Hospital, DePir,telaan, 135, B-9.000 Ghent (Belgium)
(Received August 18, 1974)
Summary Over a 10-month pt'riod, after a fight pulmonary graft in a 23 year old male, 90 serum LDH patterns were obtained; the anaerobic serum LDH-5 and LDH-4 fractions rose in acute rejection periods and during chronic rejection, together with total LDH activity. The determination of serum LDH isoenzymes may be useful to detect rejection in pulmonary transplantation.
Introduction
The assay of serum LDH isoenzymes may be of use in detecting rejection after transplantation of heart [3] or kidneys [4]. We report here the serum total LDH activity and LDH isoenzymes after a human lung homotransplantation with 10 months survival. Methods Total LDH activity was determined by the "backward reaction" method according to Warburg and Christian [8], where the decrease of extinction is analysed spectrophotometrically [2], to allow measurements at 37°C in a barbital buffer. The results are expressed in i~ ternational units per ml. LDH isoenzymes were studied by agar gel elect-ophorcsis on microscope slides according to Wieme [6,7]. Visualisation of the eiectrophoretic pattern was achieved usir.g tetrazolium blue with a modified sandwich technique .and the absorbance c f each fraction was measured using a Vitatron spectrophotometer with attached logarithmic recorder. Case report The patient was a 23 year old male with micronodular silicosis. He under-
292 TABLE I SERUM LDH ACTIVITY
IN
LUNG
HOMOTRANSPLANTATION;
LDH
ISOENZYMES
AND
TOTAL
LDH
Nov
Dec
.Jan
Feb
Mar
Apr
May
Jun
Jul
Aug
Sep
Nor- Confidence mal limits values + a n d - 2 o
21.1 22.1 19.9 18.1 18.5
24.5 29.2 21.8 15.6 6.7
22.6 22.2 20.4 19.5 14.9
20.4 26.1 21.2 20.4 11.6
17.6 24.3 22.6 21.2 14.0
18.5 25.2 22.6 19.8 13.7
18.9 2"i.8 23.5 18.8 10.8
20.3 28.7 21.0 17.0 12.6
16.8 26.7 25.6 19.2 11.4
17.8 21.6 20.6 20.1 19.3
15.9 21.9 21.9 20.6 19.7
43.6 41,1 12.1 3.2 0.0
2.8
1.9
3.4
"7
3.1
2.2
2.4
2.2
3.3
4.6
5.8
0.5
1968
1969
% LDH-1 LDH-2 LDIt-3 LDH-4 LDH-5 Total LDH activity (1.U./mi)
39--54 35----47 9---19 2--5 0--5
(0.3--
0.7)
went a right lung homotransplantation on the 14th of N o v e m b e r 1 9 6 8 and survived until the 10th of September 1969. He had an acute clinical rejection crisis on the 5th postoperative day, another one in January 1 9 6 9 and he finally aied having presented clinical s y m p t o m s o f chronic rejection during the last 2 months of his life [ 1 ]. Results In this 10-month period 90 determinations of serum total LDH activity and LDH isoenzymes were made. The mean results for each m o n t h are presented in Table I. Table II shows the LDH patterns we observed in t w o normal-looking specimens of human lung tissue obtained from t w o p~tients during operations for bronchial carcinoma. It is clear that the a m o u n t of anaerobic M units in lung tissue is much higher than in heart muscle tissue or renal cortex. Discussion The total serum LDH activity was never normal during the entire observa. T A B L E II LDI1 I S O E N Z Y M E
PAT'rERN
IN H U M A N L U N G T I S S U E
L I ) H i s o e n z y m e (%)
LDH-1 LDH-2 LDH-3 LDH-4 LDH-5
Spet:imen I
Specitnen H
6.7 19.6 25.9 24.7 25.1
4.3 20.3 37.9 27.5 9.9
293
tion time. The lowest figures were observed ~n December 1968, April and June 1969; in these periods the patient was doing well clinically. The total serum LDH leve! reached peak values in the immediate postoperative phase, and -with an acute rejection period at the 5th day. High values were seen in January, 1969, a month where an episode of dyspnoea occurred for which the immunosuppressive drug regimen had to be increased. In the last 3 months the total serum LDH activity rose from 3.3 over 4.6 to 5.8 I.U./ml together with a deteriorating clinical condition. The serum LDH-5 and LDH-4 increased during the entire 10-month course after transplantation. This increase is n o t only a percentual one but also an absolute one as the total LDH activity was very high. The aerobic LDH-1 and LDH-2 figures were very low during the same period. As the lung contains more anaerobic M units than aerobic H units P ralease of LDH from a rejected lung might provoke a serum increase of the ana,~robic LDH-5 and LDH-4 fractions. The serum LDH.5 was very high in November 1968, and August and September 1969; these periods correspond, respectively, to an acute clinical rejection episode and to increasing chronic rejection in the months before his death. Low or almost normal serum LDH-5 was present in December 1968, when the patient was doing well clinically. From December to January the serum LDH-5 almost doubled and in January we had to treat the patient for rejection again. Thus the evolution of pulmonary graft rejection in this patient seems to bear a good correlation with the serum LDH.5 values; this becomes evident when we mention the serum LDH-5 increase from 7% on the 4th postoperative day to 17% on the 5th postoperative day, where we had all the clinical symptoms of graft rejection with dyspnoea, fever, fall of pO: and good response on an adequate increase oi immunosuppressive therapy. The persistent high total serum LDH activity and increased anaerobic fractions over the entire follow-up period carried a bad prognosis for the graft and for the patient. We do not think a toxic effect of azathioprinc on the i.;ver could be responsible for the observed serum LDH isoenzyme changes [5] as there was never an increase of serum transaminases, and as there were no hepatitis lesions on autopsy. Conclusion
In this case of pulmonary transplantation in man, the assay of serum LDH isoenzymes and total serum LDH activity r, as been useful in diagnosis of acute rejection and in following chronic rejection, an increa~ of LDH-5 and LDH-4 together with a total LDH activity increase indicating rejection of the graft. Acknowledgement Supported by the Belgian Fonds Voor Wetenschappelijk Geneeskv-dig Onderzoek.
294
References 1 2 3 4 5 6 7 8
F. Derom, F, Barbier ~nd S. Ringoir, J. Thorae. Cardiovasc. Surg., 61 ( 1 9 7 1 ) 835 F. K u b o w i t z and P. Ott, Bioehem. Z.. 314 ( 1 9 4 3 ) 94 J.J. Nora. D.A. Cooley and D.J. Fernbach, New Engl. J. Med., 280 ( 1 9 6 9 ) 1079 S. Ringoir° R.J, Wieme and F. Derom, E x c e r p t a Medica Int. Congress series. No. 179, Proc. E.D.T.A. Dublin. 1968, p. 270 T,E. Starzl. T.L. Marchioro and K.A. Porter, j u : g e r y , ,58 (19C5) 131 R.J. Wieme, Ctin. Chim. Acta, 4 (1959) 317 R.J. Wieme, Aoargel Electrophoresis. T h e o r e t i c a l and Applied, Elsevier, A m s t e r d a m ° 1965 O. W a r b u ~ a id W. Christian, Biochem. Z., 186 (1963) 81
