- Email: [email protected]
L-ASPARAGINASE AND IMMUNOGLOBULINS
1364 contains monosodium and disodium hydrogen phosphates in the reverse of the 4 to 1 ratio in the oral neutral phosphate used by the Australian workers. Both parties seem to have been unaware of this discrepancy, which may account for some of the disagreement. Also, the cation calcium was conspicuously ignored. Calcium-phosphate salts would be highly undesirable. ARTHUR G. SCHOCH.
BLOOD-GROUPS AND INFLUENZA
SIR, Three investigations in Britain have revealed varying susceptibility to influenza among people with different ABO blood-groups. McDonald and Zuckermanfound a relative excess of blood-group-0 patients among 2000 R.A.F. personnel treated in hospital for proved influenzaA2 infections as compared to the blood-group distribution in 47,000 R.A.F. recruits tested over the same period. Potter and Schildreported that, in 505 sera, hxmagglutination-inhibition (R.!.) antibody to A2/Singapore influenza was more prevalent in group-0 individuals than in people with other blood-groups, but the difference was statistically TABLE I-GROUPS STUDIED AND INFECTION-RATES IN SERA BEFORE,
*
Indicated by
a
DURING, AND
4-fold
or
AFTER DEFINED EXPERIENCE
greater rise in titre between lst and 2nd
greater rise in H.I. titre) before and at the end of defined experiences (table i). Analysis of observed and expected infection-rates according to ABO blood-groups revealed no significant differences by the X2 test for the four groups, taken individually or combined (table II). Antibody to influenza A2 virus was already present in many sera from three of the four groups on the first bleeding, but neither the prevalence-rates for antibody at this time nor the subsequent incidence-rates in those lacking antibody revealed significant differences in susceptibility according to ABO blood-type. Previous immunity played no role in the fourth group (Yale students) because they were observed during the course of the A2/Hong Kong epidemic to which they had not hitherto been exposed. or
Clinical data were available for the Yale students. These revealed no ABO blood-type differences when the attackrate for proved clinical illness was measured on the basis of the entire population at risk or when measured on the basis of the frequency of clinical illness among those infected. Similar results were obtained for influenza B and parainfluenza 1-3 infections, but the infection-rates were much lower. The details of this and other work on viral disease will be published later. W.H.O. Serum Reference Bank, Department of Epidemiology and Public Health, Yale University School of Medicine, New Haven, Connecticut 06510.
L-ASPARAGINASE AND IMMUNOGLOBULINS SIR,-Numerous reports have shown that L-asparaginase (Asn-ase) has a suppressant effect on cellular immunity,1-7 Furthermore, the reduction of antibody-forming cells and antibody titres in actively immunised animals treated with Asn-ase suggests that the enzyme also suppresses humoral immunity 8-13; on the other hand, in the spleens of rabbits injected with Asn-ase only the paracortical areas were reduced in size, whereas the germinal centres appeared
unchanged
samples. TABLE II-SUMMARY OF
A2-INFECTION
RATES
ACCORDING TO BLOOD-TYPE
ALFRED S. EVANS KATHLEEN SHEPARD VIRGINIA RICHARDS.
or
enlarged.’-’
It has been suggested that Asn-ase should be tried in autoimmune diseases.15 We wish to report briefly the behaviour of the main serum-immunoglobulins in children treated with Asn-ase. IgG, IgA, and IgM were measured
using Hyland’Immunoplates ’ (all sera were stored at - 20°C before testing). 8 children were investigated before, during, and after Asn-ase (’ Crasnitin ’) treatment. 6 of them (cases 1-6 in the accompanying table) had acute lymphocytic leukaemia and 2 (cases 7 and 8) had rheumatoid arthritis. Their ages ranged from 3 to 12 years. Cases 1-3 had not been treated Obs. = no. observed.
Exp. =no. expected on basis of attack-rate of 373/926 Z2= 1-60 (not significant at 5 % level).
different
or
40.3%.
for the 16-20 and the over-30 age-groups. awl. inoculated volunteers with various influenza strains and noted a higher infection-rate among those with blood-group 0; but, of the infected group, more with blood-group A developed symptoms. In addition, an American study of FM1 and Asian antibody in the sera of military recruits showed that the prevalence-rates were higher in blood-type-O and blood-type-B individuals than in those with blood-type A.4 We have analysed influenza-virus infection-rates according to blood-type prospectively in four groups of young adults. Sera were tested for infection (as shown by a 4-fold
Tyrrell
1. 2. 3. 4.
only
before. Cases 4-6 had been treated with other antileukæmic drugs, but all treatment was stopped 6-7 days before Asn-ase was begun. Cases 7 and 8 had been unresponsive to conventional rheumatoid-arthritis treatment, and their parents consented to the use of Asn-ase. 1.
et
McDonald, J. C., Zuckerman, A. J. Br. med. J. 1962, ii, 89. Potter, C. W., Schild, G. C. J. Immun. 1967, 98, 1320. Tyrrell, D. A. J., Sparrow, P., Beare, A. S. Nature, 1968, 220, 819. Cuadrado, R., Davenport, F. M. Bull. Wld Hlth Org. 1970, 42, 873.
2. 3. 4. 5. 6. 7.
8. 9. 10. 11. 12. 13. 14. 15.
Astaldi, G., Burgio, G. R., Krč, I., Genova, R., Astaldi, A., Jr. Lancet, 1969, i, 423. Weiner, M. S., Waithe, W. I., Hirschhorn, K. ibid. 1969, ii, 748. Schulten, H. K., Giraldo, G., Boyse, E. A., Oettgen, H. F. ibid. p. 644. Ohno, R., Hersch, E. M. Blood, 1970, 35, 250. Hobib, W. P. Naturwissenschaften, 1969, 56, 217. Bertelli, A., Donati, L., Trabucchi, E., Jr. Arch. ital. Pat. 1969, 11, 475. Burgio, G. R., Astaldi, A., Jr., Krc, I., Micu, D., Astaldi, G. Rec. Results Cancer Res. 1970, 33, 288. Schwartz, R. S. Nature, 1969, 224, 275. Muller-Berat, C. N. Acta path. microbiol. scand. 1969, 77, 750. Berenbaum, M. C. Nature, 1970, 225, 550. Chakrabarty, A. K., Friedman, H. Science, 1970, 167, 869. Madaus, W. P. Klin. Wschr. 1970, 48, 240. Emmerling, P., Finger, H. Dt. med. Wschr. 1970, 95, 1578. Astaldi, G., Micu, D., Astaldi, A., Jr., Burgio, G. R. Blut (in the press). Khan, A., Hill, J. M., Adachi, M. J. Immun. 1970, 256, 105.
1365 IMMUNOGLOBULIN LEVELS
*
BEFORE, DURING,
AND AFTER
ASN-ASE*
Patients 1, 2, 3, and 7 received penicillin every 15 days; patient 7 was given aspirin (80 mg./kg./day); case 2 received 2 blood-transfusions (150+ 150 ml. on the llth day and on the 22nd day); case 3 received 1 blood-transfusion (150 ml. on 10th day).
Different dosages the treatment well tolerated.
was
were
used, and in
cases
very short because the
drug
4 and 8 was not
The results obtained show that often the
immunoglobulin values increased. IgG and IgM increased by 25% or more in 5 out of 8 and in 6 out of 8 patients respectively; IgA increased by 25% or more in all. It was exceptional for immunoglobulin values to fall. We think it unlikely that the changes in immunoglobulin levels were caused by the collateral treatment (used only in cases 1, 2, 3, and 7). It seems more likely that Asn-ase, at the dosages employed, acted as an antigen. 16, 17 Possibly the enzyme also produces nonspecific immunological stimulation or favours release of immunoglobulins into the bloodstream. Pædiatric Clinic, University of Pavia, and Blood Research Foundation Centre,
Tortona, Italy.
G. R. BURGIO R. VACCARO M. CLARA GASPARONI A. ASTALDI, JR.
C 57B 1 /6 mice were divided into three groups. Group 1 received intraperitoneal injections of 1000 I.U. L-asparaginase per kg. daily. After7 days’ treatment a skin allograft from a DBA/2 donor was applied, and L-asparaginase treatment was continued daily until the graft was rejected. Group 2 received a skin allograft from a DBA/2 donor. 1000 i.u. L-asparaginase per kg. was given intraperitoneally 24 hours after grafting and continued daily until graft rejection. Group 3 received normal saline solution (0-4 ml. per mouse) for 7 days. On day 8, a skin allograft from a DBA/2 donor was applied and daily saline treatment continued until the graft was rejected. There was only slight prolongation of graft survival when L-asparaginase was given before the application of the allograft. L-Asparaginase given after the application of an allograft showed no significant increase in survival over saline controls (see accompanying table). INFLUENCE OF L-ASPARAGINASE ON GRAFT
REJECTION
L-ASPARAGINASE AND ALLOGRAFT IMMUNITY
SIR The inhibitory effect of L-asparaginase on tumours is well established,18 and it has been shown that Escherichia coli L-asparaginase suppresses phytohæmagglutinin-induced blastogenesis.19-21 In addition, L-asparaginase considerably suppresses the humoral response 22-24 ; and in one reported case 24 injection of high doses slightly suppressed skin-graft rejection. We have investigated the effects of conventional doses of L-asparaginase on allograft survival in mice. 16. Khan, A., Hill, J. M. J. Lab. clin. Med. 1969, 73, 846. 17. Beard, M. E. J. Br. med. J. 1970, i, 191. 18. Old, L. J., Boyse, E. A., Campbell, H. A., Bradley, R. S., Fidler, J., Teller, J. D. Cancer, N. Y. 1967, 20, 1066. 19. Astaldi, G., Burgio, G. R., Krc, J., Genova, R., Astaldi, A. A. Lancet, 1969, i, 423. 20. McElwain, T. J., Hayward, S. K. ibid. p. 527. 21. Astaldi, G., Burgio, G. R., Biscotti, G., Astaldi, A., Ferfoglio, L. ibid. 1969, ii, 275. 22. Schwartz, R. S. Nature, 1969, 224, 275. 23. Chakrabarty, A. K., Friedman, H. Science, N.Y. 1970, 167, 869. 24. Nelson, S. D., Lee, M. B., Bridges, J. M. Transplantation, 1970, 9, 566.
All animals grafted on day 8. † Mean ± standard deviation. ‡As determined by Student’s test. § Not significant. a related investigation, the im--nunosuppressive effects L-asparaginase were evaluated in canine cardiac allografts. The hearts of mongrel dogs were reimplanted in the necks of 2 recipient dogs. Restoration of the circulation was accomplished by anastomoses between the innominate and subclavian arteries of the donor heart with the proximal and distal segments of the carotid arteries of the recipients.25 Daily intravenous injections of L-asparaginase (1000 i.u. per kg.) were begun 5 days before transplantation and continued until the heart ceased to contract (rejection).
In
of
25.
Tennenbaum, J. I., St. Pierre, R. L., Vasko, J. S. Archs Surg. 1969, 99, 753.
BLOOD-GROUPS AND INFLUENZA
SIR, Three investigations in Britain have revealed varying susceptibility to influenza among people with different ABO blood-groups. McDonald and Zuckermanfound a relative excess of blood-group-0 patients among 2000 R.A.F. personnel treated in hospital for proved influenzaA2 infections as compared to the blood-group distribution in 47,000 R.A.F. recruits tested over the same period. Potter and Schildreported that, in 505 sera, hxmagglutination-inhibition (R.!.) antibody to A2/Singapore influenza was more prevalent in group-0 individuals than in people with other blood-groups, but the difference was statistically TABLE I-GROUPS STUDIED AND INFECTION-RATES IN SERA BEFORE,
*
Indicated by
a
DURING, AND
4-fold
or
AFTER DEFINED EXPERIENCE
greater rise in titre between lst and 2nd
greater rise in H.I. titre) before and at the end of defined experiences (table i). Analysis of observed and expected infection-rates according to ABO blood-groups revealed no significant differences by the X2 test for the four groups, taken individually or combined (table II). Antibody to influenza A2 virus was already present in many sera from three of the four groups on the first bleeding, but neither the prevalence-rates for antibody at this time nor the subsequent incidence-rates in those lacking antibody revealed significant differences in susceptibility according to ABO blood-type. Previous immunity played no role in the fourth group (Yale students) because they were observed during the course of the A2/Hong Kong epidemic to which they had not hitherto been exposed. or
Clinical data were available for the Yale students. These revealed no ABO blood-type differences when the attackrate for proved clinical illness was measured on the basis of the entire population at risk or when measured on the basis of the frequency of clinical illness among those infected. Similar results were obtained for influenza B and parainfluenza 1-3 infections, but the infection-rates were much lower. The details of this and other work on viral disease will be published later. W.H.O. Serum Reference Bank, Department of Epidemiology and Public Health, Yale University School of Medicine, New Haven, Connecticut 06510.
L-ASPARAGINASE AND IMMUNOGLOBULINS SIR,-Numerous reports have shown that L-asparaginase (Asn-ase) has a suppressant effect on cellular immunity,1-7 Furthermore, the reduction of antibody-forming cells and antibody titres in actively immunised animals treated with Asn-ase suggests that the enzyme also suppresses humoral immunity 8-13; on the other hand, in the spleens of rabbits injected with Asn-ase only the paracortical areas were reduced in size, whereas the germinal centres appeared
unchanged
samples. TABLE II-SUMMARY OF
A2-INFECTION
RATES
ACCORDING TO BLOOD-TYPE
ALFRED S. EVANS KATHLEEN SHEPARD VIRGINIA RICHARDS.
or
enlarged.’-’
It has been suggested that Asn-ase should be tried in autoimmune diseases.15 We wish to report briefly the behaviour of the main serum-immunoglobulins in children treated with Asn-ase. IgG, IgA, and IgM were measured
using Hyland’Immunoplates ’ (all sera were stored at - 20°C before testing). 8 children were investigated before, during, and after Asn-ase (’ Crasnitin ’) treatment. 6 of them (cases 1-6 in the accompanying table) had acute lymphocytic leukaemia and 2 (cases 7 and 8) had rheumatoid arthritis. Their ages ranged from 3 to 12 years. Cases 1-3 had not been treated Obs. = no. observed.
Exp. =no. expected on basis of attack-rate of 373/926 Z2= 1-60 (not significant at 5 % level).
different
or
40.3%.
for the 16-20 and the over-30 age-groups. awl. inoculated volunteers with various influenza strains and noted a higher infection-rate among those with blood-group 0; but, of the infected group, more with blood-group A developed symptoms. In addition, an American study of FM1 and Asian antibody in the sera of military recruits showed that the prevalence-rates were higher in blood-type-O and blood-type-B individuals than in those with blood-type A.4 We have analysed influenza-virus infection-rates according to blood-type prospectively in four groups of young adults. Sera were tested for infection (as shown by a 4-fold
Tyrrell
1. 2. 3. 4.
only
before. Cases 4-6 had been treated with other antileukæmic drugs, but all treatment was stopped 6-7 days before Asn-ase was begun. Cases 7 and 8 had been unresponsive to conventional rheumatoid-arthritis treatment, and their parents consented to the use of Asn-ase. 1.
et
McDonald, J. C., Zuckerman, A. J. Br. med. J. 1962, ii, 89. Potter, C. W., Schild, G. C. J. Immun. 1967, 98, 1320. Tyrrell, D. A. J., Sparrow, P., Beare, A. S. Nature, 1968, 220, 819. Cuadrado, R., Davenport, F. M. Bull. Wld Hlth Org. 1970, 42, 873.
2. 3. 4. 5. 6. 7.
8. 9. 10. 11. 12. 13. 14. 15.
Astaldi, G., Burgio, G. R., Krč, I., Genova, R., Astaldi, A., Jr. Lancet, 1969, i, 423. Weiner, M. S., Waithe, W. I., Hirschhorn, K. ibid. 1969, ii, 748. Schulten, H. K., Giraldo, G., Boyse, E. A., Oettgen, H. F. ibid. p. 644. Ohno, R., Hersch, E. M. Blood, 1970, 35, 250. Hobib, W. P. Naturwissenschaften, 1969, 56, 217. Bertelli, A., Donati, L., Trabucchi, E., Jr. Arch. ital. Pat. 1969, 11, 475. Burgio, G. R., Astaldi, A., Jr., Krc, I., Micu, D., Astaldi, G. Rec. Results Cancer Res. 1970, 33, 288. Schwartz, R. S. Nature, 1969, 224, 275. Muller-Berat, C. N. Acta path. microbiol. scand. 1969, 77, 750. Berenbaum, M. C. Nature, 1970, 225, 550. Chakrabarty, A. K., Friedman, H. Science, 1970, 167, 869. Madaus, W. P. Klin. Wschr. 1970, 48, 240. Emmerling, P., Finger, H. Dt. med. Wschr. 1970, 95, 1578. Astaldi, G., Micu, D., Astaldi, A., Jr., Burgio, G. R. Blut (in the press). Khan, A., Hill, J. M., Adachi, M. J. Immun. 1970, 256, 105.
1365 IMMUNOGLOBULIN LEVELS
*
BEFORE, DURING,
AND AFTER
ASN-ASE*
Patients 1, 2, 3, and 7 received penicillin every 15 days; patient 7 was given aspirin (80 mg./kg./day); case 2 received 2 blood-transfusions (150+ 150 ml. on the llth day and on the 22nd day); case 3 received 1 blood-transfusion (150 ml. on 10th day).
Different dosages the treatment well tolerated.
was
were
used, and in
cases
very short because the
drug
4 and 8 was not
The results obtained show that often the
immunoglobulin values increased. IgG and IgM increased by 25% or more in 5 out of 8 and in 6 out of 8 patients respectively; IgA increased by 25% or more in all. It was exceptional for immunoglobulin values to fall. We think it unlikely that the changes in immunoglobulin levels were caused by the collateral treatment (used only in cases 1, 2, 3, and 7). It seems more likely that Asn-ase, at the dosages employed, acted as an antigen. 16, 17 Possibly the enzyme also produces nonspecific immunological stimulation or favours release of immunoglobulins into the bloodstream. Pædiatric Clinic, University of Pavia, and Blood Research Foundation Centre,
Tortona, Italy.
G. R. BURGIO R. VACCARO M. CLARA GASPARONI A. ASTALDI, JR.
C 57B 1 /6 mice were divided into three groups. Group 1 received intraperitoneal injections of 1000 I.U. L-asparaginase per kg. daily. After7 days’ treatment a skin allograft from a DBA/2 donor was applied, and L-asparaginase treatment was continued daily until the graft was rejected. Group 2 received a skin allograft from a DBA/2 donor. 1000 i.u. L-asparaginase per kg. was given intraperitoneally 24 hours after grafting and continued daily until graft rejection. Group 3 received normal saline solution (0-4 ml. per mouse) for 7 days. On day 8, a skin allograft from a DBA/2 donor was applied and daily saline treatment continued until the graft was rejected. There was only slight prolongation of graft survival when L-asparaginase was given before the application of the allograft. L-Asparaginase given after the application of an allograft showed no significant increase in survival over saline controls (see accompanying table). INFLUENCE OF L-ASPARAGINASE ON GRAFT
REJECTION
L-ASPARAGINASE AND ALLOGRAFT IMMUNITY
SIR The inhibitory effect of L-asparaginase on tumours is well established,18 and it has been shown that Escherichia coli L-asparaginase suppresses phytohæmagglutinin-induced blastogenesis.19-21 In addition, L-asparaginase considerably suppresses the humoral response 22-24 ; and in one reported case 24 injection of high doses slightly suppressed skin-graft rejection. We have investigated the effects of conventional doses of L-asparaginase on allograft survival in mice. 16. Khan, A., Hill, J. M. J. Lab. clin. Med. 1969, 73, 846. 17. Beard, M. E. J. Br. med. J. 1970, i, 191. 18. Old, L. J., Boyse, E. A., Campbell, H. A., Bradley, R. S., Fidler, J., Teller, J. D. Cancer, N. Y. 1967, 20, 1066. 19. Astaldi, G., Burgio, G. R., Krc, J., Genova, R., Astaldi, A. A. Lancet, 1969, i, 423. 20. McElwain, T. J., Hayward, S. K. ibid. p. 527. 21. Astaldi, G., Burgio, G. R., Biscotti, G., Astaldi, A., Ferfoglio, L. ibid. 1969, ii, 275. 22. Schwartz, R. S. Nature, 1969, 224, 275. 23. Chakrabarty, A. K., Friedman, H. Science, N.Y. 1970, 167, 869. 24. Nelson, S. D., Lee, M. B., Bridges, J. M. Transplantation, 1970, 9, 566.
All animals grafted on day 8. † Mean ± standard deviation. ‡As determined by Student’s test. § Not significant. a related investigation, the im--nunosuppressive effects L-asparaginase were evaluated in canine cardiac allografts. The hearts of mongrel dogs were reimplanted in the necks of 2 recipient dogs. Restoration of the circulation was accomplished by anastomoses between the innominate and subclavian arteries of the donor heart with the proximal and distal segments of the carotid arteries of the recipients.25 Daily intravenous injections of L-asparaginase (1000 i.u. per kg.) were begun 5 days before transplantation and continued until the heart ceased to contract (rejection).
In
of
25.
Tennenbaum, J. I., St. Pierre, R. L., Vasko, J. S. Archs Surg. 1969, 99, 753.