- Email: [email protected]
OSCILLATION DURING THE IMMUNE RESPONSE
475
during this period. In summary, 3 out of 4 patients H.H.T. experienced the appearance, enlargement, or aggravation of bleeding of lesions during oral contraceptive therapy. The appearance of telangiectasias in H.H.T. at puberty, and the correlation of epistaxes with the menstrual cycle, give weight to a possible causal relationship between œstrogens and the aggravation of H.H.T. Paradoxically, oral cestrogens have been recommended to counteract epistaxes in this condition.1,2 We hope that our experience will stimulate the prospective study of additional patients to bled with
determine whether in fact women with H.H.T. represent a group genetically predisposed to increased morbidity from oral contraceptives. PETER T. ROWLEY Department of Medicine, Stanford University School of Medicine, JOHN KURNICK RICHARD CHEVILLE. Palo Alto, California.
MITOCHONDRIAL ASPARTATE TRANSAMINASE SIR,-It has recently become possible to determine the isoenzymes of aspartate transaminase (G.O.T.) by a sensitive semiquantitative method,3 and this technique has now been applied to a series of African patients, mostly from the wards of Mulago Hospital. As well as seeking out patients suffering from various individual diseases, we have obtained samples from a wide variety of cases-as nearly as possible a whole medical-ward population over a period of several days. It is known that serum levels of both isoenzymes, but predominantly of the cytoplasmic form, are raised in infective hepatitis 4; and following myocardial infarction it is the mitochondrial form which shows a greater relative increase. A striking feature of the present cases is that many show a considerable rise in the level of mitochondrial isoenzyme alone, or have only a slight change in the level of cytoplasmic isoenzyme. Of nine with grade D or E mitochondrial isoenzyme (over 15 I.U. per litre) and not more than 23 I.U. per litre total aspartate transaminase,6 three have cirrhosis of the liver, three have schistosomiasis, one has hepatitis, one has primary hepatoma, and one has had treatment for malignant melanoma but was admitted on this occasion with an inflammatory mass in the right iliac fossa which had almost resolved when the blood-sample was taken. All four confirmed cases of primary hepatoma (including the one noted above) and several suspected cases, show a considerable rise in the level of mitochondrial enzyme (grade c or above), with not more than 21 I.U. per litre total aspartate transaminase. So far, only two cases of endomyocardial fibrosis have been tested: both show distinct elevation of the mitochondrial enzyme. It has been reportedthat the total-aspartate-transaminase level is usually normal in this condition, but the method used6 is very insensitive to the mitochondrial isoenzyme.5 A large number of other cases show modest increases of the mitochondrial isoenzyme alone, or increases of both together, but we will not discuss these further here. We do not wish any premature conclusions to be drawnfor example, in cases of endomyocardial fibrosis it will be difficult to ascertain whether excess serum enzymes arise from the heart or the liver, and even more difficult to 1.
Koch, H. J., Escher, G. C., Lewis, J. S. J. Am. med. Ass. 1952, 149,
1376. 2. Harrison, D. F. Q. Jl Med. 1964, 33, 25. 3. Boyde, T. R. C. Z. klin. Chem. 1968, 6, 431. 4. Villa, L., Dioguardi, N., Agostoni, A. Klin. Wschr. 5. Boyde, T. R. C. Enzym. biol. clin. 1968, 9, 385.
1967, 45, 44.
6. Reitman, S., Frankel, S. Am. J. clin. Path. 1957, 28, 56. 7. Campbell, J., Somers, K. Br. med. J. 1960, i, 1540.
determine the mechanism of release. It seems clear, of mitochondrial enzymes in the serum will add a new dimension to clinical enzymology, by allowing an approach to the study of intracellular events through observations on the blood.
however, that study
Clinical Biochemistry Laboratory,
Mulago Hospital. Biochemistry Department Makerere University College, P.O. Box 7072, Kampala, Uganda.
S. N. FARMER. A. M. NANJI T. R. C. BOYDE.
OSCILLATION DURING THE IMMUNE RESPONSE SIR,—The report by Dr. Dwyer and Dr. Mackay (Jan. 24, p. 164), that the number of antigen-binding lymphocytes in blood shows rapid fluctuation after antigenic stimulation, adds to the evidence that oscillation may occur during the immune responses. 1-3 The factors responsible for oscillation in the production of red-cells, granulocytes, and platelets have lately been reviewedand, in view of the close similarity between the production of differentiated blood-cells and antibody-producing cells, it may be questioned whether similar factors operate during the response to antigenic stimulation. Two principal mechanisms may give rise to oscillation in biological systems-negative feedback and interaction between populations. There is now much evidence that antibody controls its own production by some form of negative feedback.5 Britton and Moller1 have reported that the number of 19S-antibody-forming cells in mice showed oscillation following the administration of endotoxin. Since these workers were able to prevent a rise in the number of these cells by administering antibody beforehand, their observations provide strong evidence that negative feedback was responsible for the oscillation. The point raised by Dr. Dwyer and Dr. Mackay, that the fluctuations observed in their own study were too rapid to be accounted for by negative feedback, is not necessarily valid, since rapidity of oscillation in a feedback system may merely indicate that the time-delay in or time-characteristic of the system is brief. The possible role of interaction between populations is difficult to assess and quite speculative. In their classic early work Lotka and Volterra pointed out the oscillatory nature of the equilibrium which results from one type of interaction between populations-that between two animal species, one of which is the predator and one of which is the prey. This type of Lotka-Volterra equilibrium may be involved in the interaction between replicating and immune cells (the predators). Similar types of interaction, however, may also be involved in the competition for stem-cells or for an antigen between different antibody-producing cell-lines, or in the interaction between a stem-cell compartment and a more differentiated compartment. Oscillation is more than a phenomenon of visual interest. Its presence may point to the existence of hitherto unsuspected feedback loops or population interactions. Conversely, the ubiquity of feedback loops and population interactions makes it possible that many physiological
antigen (the prey)
Britton, S., Moller, G. J. Immun. 1968, 100, 1326. Radovich, J., Hemingsen, H., Talmage, D. W. ibid. 1969, 102, 288. Cornelius, E. A., Yunis, E. J., Martinez, C. Proc. Soc. exp. Biol. Med. 1969, 131, 680. 4. Morley, A. A., King-Smith, E. A., Stohlman, F. Jr. in Hemopoietic Cell Proliferation (edited by F. Stohlman, Jr.). New York (in the press). 5. Uhr, J. W., Moller, G. Adv. Immun. 1968, 8, 81. 6. Lotka, A. J. Elements of Physical Biology; p. 88 Baltimore, 1925. 7. Volterra, V. Biotheorenca, 1937, 3, 1.
1. 2. 3.
476 systems show temporal organisation. With respect to the immune response, many disorders in which disturbed immunity is involved or suspected to be involved are characterised by fluctuations in their course. Perhaps many of these fluctuations are due to the inherent rhythmicity of the immune system rather than to the action of unrecognised external factors. ALEC MORLEY St. Elizabeth’s Hospital, FREDERICK STOHLMAN, JR. Boston, Mass. 02135.
STEROID THERAPY FOR HÆMOPHILLA
SIR,—Mr. Giordano and Dr. Radivoyevitch (Jan. 31, p. 249) present data showing a seasonal variation in plasmacortisol levels in 15 patients with haemophilia. They suggest that haemophiliacs may benefit from steroid treatment. In 1967 we1 published a report of a controlled trial of treatment of hxmophilia with steroids in children and adults. We showed that the number of bleeding events was significantly reduced in children, but not in adults, by continuous administration of prednisolone (children 3 mg. daily; adults 7-5 mg. daily). However, the severity of the disorder did not seem to be reduced; for we measured the disability resulting from the disease by the number of days off sick and number of days in bed, and these were not altered. Our findings thus suggested that only the more trivial bleeding events were suppressed by this treatment, which could not, therefore, be recommended in view of the possible disadvantages of long-term steroid administration. Departments of Social Medicine and Hæmatology, St. Thomas’s Hospital A. E. BENNETT Medical School, G. I. C. INGRAM. London S.E.1.
MARBURG VIRUS the occurrence of fatal human cases of SIR,-In 1967 disease in Germany and Yugoslavia, resulting heemorrhagic from contact with African vervet monkeys, stimulated intensive investigations in laboratories in several countries including Germany, England, the United States, and South Africa, where infective material had been received. A new type of agent, now generally called the Marburg virus, was isolated; and its properties have been studied to some extent. It was not clear where the infection originated. The monkeys had been sent from Uganda by air, but had undergone an overnight stop at London in a room containing numerous other animals including birds and reptiles. British investigators2 failed to find complement-fixing antibody to Marburg virus in over 200 vervet-monkey sera, most of which came from animals caught in Uganda at about the same time as the monkeys which caused the disease in Europe. In January, 1969, however, a report3 from the United States averred that sera from a high proportion of primates born in Africa showed complement-fixing antibody to Marburg virus. This disturbing news led us to find out whether a similar situation existed in South Africa. Between March and June, 1969, we tested sera from 268 baboons (Papio ursinus) and 211 vervet monkeys (Cercopithecus œthiops pygerythrus), representing a wide geographical distribution in the country, for complement-fixing antibody. In each test we used our own live-virus antigen prepared from the liver of a laboratory infected monkey, in parallel with the &bgr;-propiolactone-inactivated guineapigBennett, A. E., Ingram, G. I. C. Lancet, 1967, i, 967. Simpson, D. I. H., Bowen, E. T. W., Bright, W. F. Lab. Anim. 1968, 2, 75. 3. Kalter, S. S., Ratner, J. J., Heberling, R. L. Proc. Soc. exp. Biol. Med. 1969, 130, 10.
1. 2.
liver antigen sent to us by the United States workers; while uninfected monkey and guineapig-liver antigens were used Care was taken to ensure that the as negative controls. testing technique was identical to that used in the United States. Our results confirmed that complement fixation occurred in a high proportion (61 %) of sera when the United States positive antigen was used; but with our own live-virus antigen only seven positive sera were found, and all of these also gave strong reactions with the negative liver antigens. Control tests with convalescent sera from four German patients, and with two hyperimmune guineapig sera sent to us from England, showed that our positive antigen possessed a higher activity against known Marburg antibody than did the United States positive antigen. These results were presented at the symposium on Marbury Virus Disease held at Marburg/Lahn, West Germany, in June, 1969. We concluded that the United States antigen probably contained a second component not specific for Marburg virus, which fixed complement in tests on simian sera. Our findings were supported by the fact that deliberate search made here since 1954 for simian viruses has not produced a single isolate of Marburg virus from wild monkeys or baboons in South Africa. We do not wish to imply that the virus cannot occur here, but we have no evidence of the extensive activity suggested by the use of the antigen prepared in the United States. Considering our results together with the negative serological findings of the British investigators, we are driven to the conclusion that the origin and distribution of the virus remain unknown. Work on this dangerous agent has been discontinued in nearly all of the few laboratories handling the live virus. Our own studies have been terminated because this Foundation’s policy does not permit the study of Marburg virus while vaccines are being prepared. Since we do not know the origin of the virus, we cannot predict where it may next be encountered; and it is regrettable that this important question is not receiving sufficient attention. Poliomyelitis Research Foundation, South African Institute M. STRICKLAND-CHOLMLEY for Medical Research, H. MALHERBE. P. O. Box 1038, Johannesburg.
SIMIAN VIRUS REFERENCE CENTRE use of nonhuman primates in biomedical research has created a need for a centre where people may seek help when faced with virological problems in these animals. A simian virus reference centre has been established at this laboratory for such a purpose. It is supported by the U.S. National Institutes of Health and by the World Health Organisation as a Collaborating Laboratory on Comparative Medicine: Simian Viruses, and has the following aims:
SIR The increasing
(1) development of a working repository for simian viruses; (2) provision of a source of reagents such as certified reference seed virus strains and specific antisera; (3) provision of consultation services, including serum survey data, on the existence of antibody to various viruses of human and simian origin in various genera and species of primates; (4) provision of diagnostic services, including the identification and characterisation of viruses for primate research-workers unable to identify isolates obtained from their primates (this would also include screening for human viruses); (5) provision of information and arrangement of exchanges of organisms between primate centres and other health organisations; (6) training of interested students in virological laboratory procedures associated with primate investigations. Further details are contained in WHO Chronicle, 1969, 112 or may be had from the undersigned.
23,
Division of Microbiology and Infectious Diseases, Southwest Foundation for Research and Education, P.O. Box 28147, San Antonio, Texas 78228.
S. S. KALTER.
during this period. In summary, 3 out of 4 patients H.H.T. experienced the appearance, enlargement, or aggravation of bleeding of lesions during oral contraceptive therapy. The appearance of telangiectasias in H.H.T. at puberty, and the correlation of epistaxes with the menstrual cycle, give weight to a possible causal relationship between œstrogens and the aggravation of H.H.T. Paradoxically, oral cestrogens have been recommended to counteract epistaxes in this condition.1,2 We hope that our experience will stimulate the prospective study of additional patients to bled with
determine whether in fact women with H.H.T. represent a group genetically predisposed to increased morbidity from oral contraceptives. PETER T. ROWLEY Department of Medicine, Stanford University School of Medicine, JOHN KURNICK RICHARD CHEVILLE. Palo Alto, California.
MITOCHONDRIAL ASPARTATE TRANSAMINASE SIR,-It has recently become possible to determine the isoenzymes of aspartate transaminase (G.O.T.) by a sensitive semiquantitative method,3 and this technique has now been applied to a series of African patients, mostly from the wards of Mulago Hospital. As well as seeking out patients suffering from various individual diseases, we have obtained samples from a wide variety of cases-as nearly as possible a whole medical-ward population over a period of several days. It is known that serum levels of both isoenzymes, but predominantly of the cytoplasmic form, are raised in infective hepatitis 4; and following myocardial infarction it is the mitochondrial form which shows a greater relative increase. A striking feature of the present cases is that many show a considerable rise in the level of mitochondrial isoenzyme alone, or have only a slight change in the level of cytoplasmic isoenzyme. Of nine with grade D or E mitochondrial isoenzyme (over 15 I.U. per litre) and not more than 23 I.U. per litre total aspartate transaminase,6 three have cirrhosis of the liver, three have schistosomiasis, one has hepatitis, one has primary hepatoma, and one has had treatment for malignant melanoma but was admitted on this occasion with an inflammatory mass in the right iliac fossa which had almost resolved when the blood-sample was taken. All four confirmed cases of primary hepatoma (including the one noted above) and several suspected cases, show a considerable rise in the level of mitochondrial enzyme (grade c or above), with not more than 21 I.U. per litre total aspartate transaminase. So far, only two cases of endomyocardial fibrosis have been tested: both show distinct elevation of the mitochondrial enzyme. It has been reportedthat the total-aspartate-transaminase level is usually normal in this condition, but the method used6 is very insensitive to the mitochondrial isoenzyme.5 A large number of other cases show modest increases of the mitochondrial isoenzyme alone, or increases of both together, but we will not discuss these further here. We do not wish any premature conclusions to be drawnfor example, in cases of endomyocardial fibrosis it will be difficult to ascertain whether excess serum enzymes arise from the heart or the liver, and even more difficult to 1.
Koch, H. J., Escher, G. C., Lewis, J. S. J. Am. med. Ass. 1952, 149,
1376. 2. Harrison, D. F. Q. Jl Med. 1964, 33, 25. 3. Boyde, T. R. C. Z. klin. Chem. 1968, 6, 431. 4. Villa, L., Dioguardi, N., Agostoni, A. Klin. Wschr. 5. Boyde, T. R. C. Enzym. biol. clin. 1968, 9, 385.
1967, 45, 44.
6. Reitman, S., Frankel, S. Am. J. clin. Path. 1957, 28, 56. 7. Campbell, J., Somers, K. Br. med. J. 1960, i, 1540.
determine the mechanism of release. It seems clear, of mitochondrial enzymes in the serum will add a new dimension to clinical enzymology, by allowing an approach to the study of intracellular events through observations on the blood.
however, that study
Clinical Biochemistry Laboratory,
Mulago Hospital. Biochemistry Department Makerere University College, P.O. Box 7072, Kampala, Uganda.
S. N. FARMER. A. M. NANJI T. R. C. BOYDE.
OSCILLATION DURING THE IMMUNE RESPONSE SIR,—The report by Dr. Dwyer and Dr. Mackay (Jan. 24, p. 164), that the number of antigen-binding lymphocytes in blood shows rapid fluctuation after antigenic stimulation, adds to the evidence that oscillation may occur during the immune responses. 1-3 The factors responsible for oscillation in the production of red-cells, granulocytes, and platelets have lately been reviewedand, in view of the close similarity between the production of differentiated blood-cells and antibody-producing cells, it may be questioned whether similar factors operate during the response to antigenic stimulation. Two principal mechanisms may give rise to oscillation in biological systems-negative feedback and interaction between populations. There is now much evidence that antibody controls its own production by some form of negative feedback.5 Britton and Moller1 have reported that the number of 19S-antibody-forming cells in mice showed oscillation following the administration of endotoxin. Since these workers were able to prevent a rise in the number of these cells by administering antibody beforehand, their observations provide strong evidence that negative feedback was responsible for the oscillation. The point raised by Dr. Dwyer and Dr. Mackay, that the fluctuations observed in their own study were too rapid to be accounted for by negative feedback, is not necessarily valid, since rapidity of oscillation in a feedback system may merely indicate that the time-delay in or time-characteristic of the system is brief. The possible role of interaction between populations is difficult to assess and quite speculative. In their classic early work Lotka and Volterra pointed out the oscillatory nature of the equilibrium which results from one type of interaction between populations-that between two animal species, one of which is the predator and one of which is the prey. This type of Lotka-Volterra equilibrium may be involved in the interaction between replicating and immune cells (the predators). Similar types of interaction, however, may also be involved in the competition for stem-cells or for an antigen between different antibody-producing cell-lines, or in the interaction between a stem-cell compartment and a more differentiated compartment. Oscillation is more than a phenomenon of visual interest. Its presence may point to the existence of hitherto unsuspected feedback loops or population interactions. Conversely, the ubiquity of feedback loops and population interactions makes it possible that many physiological
antigen (the prey)
Britton, S., Moller, G. J. Immun. 1968, 100, 1326. Radovich, J., Hemingsen, H., Talmage, D. W. ibid. 1969, 102, 288. Cornelius, E. A., Yunis, E. J., Martinez, C. Proc. Soc. exp. Biol. Med. 1969, 131, 680. 4. Morley, A. A., King-Smith, E. A., Stohlman, F. Jr. in Hemopoietic Cell Proliferation (edited by F. Stohlman, Jr.). New York (in the press). 5. Uhr, J. W., Moller, G. Adv. Immun. 1968, 8, 81. 6. Lotka, A. J. Elements of Physical Biology; p. 88 Baltimore, 1925. 7. Volterra, V. Biotheorenca, 1937, 3, 1.
1. 2. 3.
476 systems show temporal organisation. With respect to the immune response, many disorders in which disturbed immunity is involved or suspected to be involved are characterised by fluctuations in their course. Perhaps many of these fluctuations are due to the inherent rhythmicity of the immune system rather than to the action of unrecognised external factors. ALEC MORLEY St. Elizabeth’s Hospital, FREDERICK STOHLMAN, JR. Boston, Mass. 02135.
STEROID THERAPY FOR HÆMOPHILLA
SIR,—Mr. Giordano and Dr. Radivoyevitch (Jan. 31, p. 249) present data showing a seasonal variation in plasmacortisol levels in 15 patients with haemophilia. They suggest that haemophiliacs may benefit from steroid treatment. In 1967 we1 published a report of a controlled trial of treatment of hxmophilia with steroids in children and adults. We showed that the number of bleeding events was significantly reduced in children, but not in adults, by continuous administration of prednisolone (children 3 mg. daily; adults 7-5 mg. daily). However, the severity of the disorder did not seem to be reduced; for we measured the disability resulting from the disease by the number of days off sick and number of days in bed, and these were not altered. Our findings thus suggested that only the more trivial bleeding events were suppressed by this treatment, which could not, therefore, be recommended in view of the possible disadvantages of long-term steroid administration. Departments of Social Medicine and Hæmatology, St. Thomas’s Hospital A. E. BENNETT Medical School, G. I. C. INGRAM. London S.E.1.
MARBURG VIRUS the occurrence of fatal human cases of SIR,-In 1967 disease in Germany and Yugoslavia, resulting heemorrhagic from contact with African vervet monkeys, stimulated intensive investigations in laboratories in several countries including Germany, England, the United States, and South Africa, where infective material had been received. A new type of agent, now generally called the Marburg virus, was isolated; and its properties have been studied to some extent. It was not clear where the infection originated. The monkeys had been sent from Uganda by air, but had undergone an overnight stop at London in a room containing numerous other animals including birds and reptiles. British investigators2 failed to find complement-fixing antibody to Marburg virus in over 200 vervet-monkey sera, most of which came from animals caught in Uganda at about the same time as the monkeys which caused the disease in Europe. In January, 1969, however, a report3 from the United States averred that sera from a high proportion of primates born in Africa showed complement-fixing antibody to Marburg virus. This disturbing news led us to find out whether a similar situation existed in South Africa. Between March and June, 1969, we tested sera from 268 baboons (Papio ursinus) and 211 vervet monkeys (Cercopithecus œthiops pygerythrus), representing a wide geographical distribution in the country, for complement-fixing antibody. In each test we used our own live-virus antigen prepared from the liver of a laboratory infected monkey, in parallel with the &bgr;-propiolactone-inactivated guineapigBennett, A. E., Ingram, G. I. C. Lancet, 1967, i, 967. Simpson, D. I. H., Bowen, E. T. W., Bright, W. F. Lab. Anim. 1968, 2, 75. 3. Kalter, S. S., Ratner, J. J., Heberling, R. L. Proc. Soc. exp. Biol. Med. 1969, 130, 10.
1. 2.
liver antigen sent to us by the United States workers; while uninfected monkey and guineapig-liver antigens were used Care was taken to ensure that the as negative controls. testing technique was identical to that used in the United States. Our results confirmed that complement fixation occurred in a high proportion (61 %) of sera when the United States positive antigen was used; but with our own live-virus antigen only seven positive sera were found, and all of these also gave strong reactions with the negative liver antigens. Control tests with convalescent sera from four German patients, and with two hyperimmune guineapig sera sent to us from England, showed that our positive antigen possessed a higher activity against known Marburg antibody than did the United States positive antigen. These results were presented at the symposium on Marbury Virus Disease held at Marburg/Lahn, West Germany, in June, 1969. We concluded that the United States antigen probably contained a second component not specific for Marburg virus, which fixed complement in tests on simian sera. Our findings were supported by the fact that deliberate search made here since 1954 for simian viruses has not produced a single isolate of Marburg virus from wild monkeys or baboons in South Africa. We do not wish to imply that the virus cannot occur here, but we have no evidence of the extensive activity suggested by the use of the antigen prepared in the United States. Considering our results together with the negative serological findings of the British investigators, we are driven to the conclusion that the origin and distribution of the virus remain unknown. Work on this dangerous agent has been discontinued in nearly all of the few laboratories handling the live virus. Our own studies have been terminated because this Foundation’s policy does not permit the study of Marburg virus while vaccines are being prepared. Since we do not know the origin of the virus, we cannot predict where it may next be encountered; and it is regrettable that this important question is not receiving sufficient attention. Poliomyelitis Research Foundation, South African Institute M. STRICKLAND-CHOLMLEY for Medical Research, H. MALHERBE. P. O. Box 1038, Johannesburg.
SIMIAN VIRUS REFERENCE CENTRE use of nonhuman primates in biomedical research has created a need for a centre where people may seek help when faced with virological problems in these animals. A simian virus reference centre has been established at this laboratory for such a purpose. It is supported by the U.S. National Institutes of Health and by the World Health Organisation as a Collaborating Laboratory on Comparative Medicine: Simian Viruses, and has the following aims:
SIR The increasing
(1) development of a working repository for simian viruses; (2) provision of a source of reagents such as certified reference seed virus strains and specific antisera; (3) provision of consultation services, including serum survey data, on the existence of antibody to various viruses of human and simian origin in various genera and species of primates; (4) provision of diagnostic services, including the identification and characterisation of viruses for primate research-workers unable to identify isolates obtained from their primates (this would also include screening for human viruses); (5) provision of information and arrangement of exchanges of organisms between primate centres and other health organisations; (6) training of interested students in virological laboratory procedures associated with primate investigations. Further details are contained in WHO Chronicle, 1969, 112 or may be had from the undersigned.
23,
Division of Microbiology and Infectious Diseases, Southwest Foundation for Research and Education, P.O. Box 28147, San Antonio, Texas 78228.
S. S. KALTER.