- Email: [email protected]
MACROPHAGE-ELECTROPHORETIC-MOBILITY (M.E.M.) TEST FOR MALIGNANT DISEASE
627
the severity and duration of adverse circumstances. The long-term effects of such experiences depend to what extent the harmony of growth 11 is disturbed. My data on obese children show that overnutrition in the early months of life causes not only an increase in linear growth, but also an equivalent advance in skeletal maturation.12 Thus the height of obese children is normal for their bone-age, and the prognosis for height is also normal." Overnutrition later in childhood has no demonstrable effects on height The adipose organ is only or skeletal maturation.12 affected by overnutrition in the early period, but the increase in the number of cells in this event is much greater than the increase in height or skeletal maturity; thus the harmony is destroyed and these individuals will have a permanently increased complement of adipose cells. Since these findings do not apply to obesity dating from later childhood, it seems likely that the first year of life is a sensitive one for the formation of adipose cells. During such a period, cell replication is sensitive to external influences which may be beneficial (perhaps essential) or pathological. The period is also one of determination, because at its end the basic complement of cells is established and thereafter cell replication probably continues at a normal rate regardless of other circumstances. As far as the adipose organ is concerned, it seems that after the end of the sensitive period alterations in nutrition affect only cell size and not cell number. From studies in animals of experimental overnutrition 14 and undernutrition, 15 it is known that during certain periods it is more easy to impair growth and that, in general, the earlier the stress, the more This work has been confirmed severe the effects. 16 and extended by the demonstration of the importance of early nutritional experiences on the size, number, and rate of growth of adipose cells in rats.’-’,18 The effects of intrauterine malnutrition (L.F.D. children) in the present study demonstrate how adverse circumstances early in organogenesis have profound effects later, and how important timing is in growth and development. If G.H. is important for adipose-cell replication (as the present data indicate), an intermediate situation between controls and L.F.D. would be predicted for children with isolated G.H. deficiency. Such children grow normally in utero and are presumably born with a normal complement of cells. These cells do not receive a proper stimulus for replication in the sensitive period and so they lag behind the rest of development as measured by skeletal maturity, but not as much as they would have done if the stress had occurred earlier. Children who lack G.H. at a later age, however, would be expected to have passed the age when the adipose organ is
sensitive, and so harmony should not have been destroyed. Such children would be expected to have a number of adipose cells which does not differ from normal when the delay in growth induced by G.H. deficiency, and evidenced by skeletal retardation, is allowed for. The findings on the C.R. children confirm this expectation. I suggest that the adipose organ in man has a finite and well-defined sensitive period during which external
circumstances have long-term effects on cellular growth. For this organ the period of determination, when the basic complement of cells is established,
probably extends from thirty weeks of intrauterine growth through to the first nine months to a year of extrauterine growth when the skinfolds cease their rapid increase. Presumably such a model, with different timings, applies to all organs in man. I thank the Wellcome Trust and the Medical Research Council for financial assistance, and Prof. J. M. Tanner, whose unfailing advice and support have been of great help. REFERENCES 1. Winick, M. Med. Clins N. Am. 1970, 54, 1413. 2. Cheek, D. B., Hill, D. E., Cordano, A., Graham, G. G. Pediat. Res. 1970, 4, 135. 3. Brook, C. G. D. Archs Dis. Childh. 1971, 46, 182. 4. Tanner, J. M., Whitehouse, R. H. Br. med. J. 1962, i, 446. 5. Brook, C. G. D., Lloyd, J. K., Wolff, O. H. ibid. 1972, ii, 25. 6. Tanner, J. M., Thomson, A. M. Archs Dis. Childh. 1970, 45, 566. 7. Edwards, M. W., Hammond, W. H., Healy, M. J. R., Tanner, J. M., Whitehouse, R. H. Br. J. Nutr. 1955, 9, 133. 8. Durmin, J. V. G. A., Rahaman, M. M. ibid. 1967, 21, 681. 9. Tanner, J. M., Whitehouse, R. H., Healy, M. J. R. A New System for Estimating Skeletal Maturity from the Hand and Wrist. Centre Internationale de L’Enfance, Paris, 1962. 10. Hirsch, J., Gallian, E. J. Lipid Res. 1968, 9, 110. 11. Widdowson, E. M. Lancet, 1970, i, 901. 12. Brook, C. G. D. M.D. thesis, University of Cambridge, 1972. 13. Lloyd, J. K., Wolff, O. H., Whelen, W. S. Br. med. J. 1961, ii, 145. 14. Widdowson, E. M., McCance, R. A. Proc. R. Soc. 1960, B, 152, 188. 15. Widdowson, E. M., McCance, R. A. ibid. 1963, B, 158, 329. 16. Winick, M., Rosso, P. Pediat. Res. 1969, 3, 181. 17. Knittle, J. L., Hirsch, J. J. clin. Invest. 1968, 47, 2091. 18. Hirsch, J., Han, P. W. J. Lipid Res. 1969, 10, 77.
MACROPHAGE-ELECTROPHORETICMOBILITY (M.E.M.) TEST FOR MALIGNANT DISEASE An
Independent Confirmation
J. A. V. PRITCHARD W. H. SUTHERLAND
J. L. MOORE C. A. F. JOSLIN
Tenovus Laboratories, Velindre
Hospital,
Whitchurch, Cardiff
Sum ary
The test
validity of a new in-vitro bloodfor cancer, which depends on the
sensitisation of the patient’s lymphocytes to a common antigen apparently present in human tumours, has been independently confirmed. Introduction
Field and Casparyreported that lymphocytes from patients with malignant disease can be stimulated by encephalitogenic factor2 (E.F.) (a basic protein derived from the human brain) to release a macrophage-slowing factor (M.S.F.) which reduces the electrophoretic mobility of guineapig macrophages. With few exceptions, the effect was not found with lymphocytes from non-malignant controls, and therefore the technique was presented as a possible in-vitro blood-test for cancer. The results could best be explained on the basis of a common antigen present in human tumours. These unexpected findings required confirmation, and we performed a similar investigation. Materials and Methods Approximately 15 ml. of venous blood was collected from healthy hospital staff and from patients with known
628 TABLE
I-TYPICAL
RECORD
OF
TIMINGS
FROM
TWO
SAMPLES
that sample. Typical timings from two samples, one " control " and one " malignant ", are shown in table I. The frequency of " wrong " pairs decreased with increasing experience of the technique and apparatus, until only about 1 cell in every 100 from control samples gave a " wrong " pair of timings. Results were expressed as a percentage change in mobility, 100 (T—c)/c, where T is the average migration time for macrophages in the test sample containing lymphocytes, macrophages, and antigen, and c is the average migration time for macrophages in the control sample containing lymphocytes and macrophages but no
antigen. Results
Between May, 1971, and July, 1972, 309 bloodsamples from patients and from healthy subjects were examined with the macrophage-electrophoretic-mobilTABLE II-PERCENTAGE SLOWING OF MACROPHAGES FROM
SUBJECTS
WITH MALIGNANT DISEASE AGAINST E.F. ANTIGEN
*
This represents
a
t This represents
a
2-7% decrease in mean migration time. 20-6% increase in mean migration time.
After defibrination, lymphocytes were harvested either by the methylcellulose technique described by Hughes and Caspary3 or by means of a ’FicolTriosil ’ technique which will be described fully elsewhere. The washed lymphocytes from either technique were Macrophages were resuspended in’Medium 199’. prepared from the peritoneal exudate of 400-600 g. Hartley albino guineapigs four to ten days after the injection of 20 ml. of sterile liquid paraffin into the peritoneum. The cells were washed and resuspended in medium 199 to a concentration of 10’ cells per ml. and then irradiated with a dose of 100-200 rad from a cxsium-137 source. The encephalitogenic factor (E.F.) was prepared from a human brain by the methods described by Caspary and Field.2,4 1-0 mg. of E.F. was dissolved in medium 199 so that the final concentration was 100 (kg. per 0-1 ml.
malignant disease.
Electrophoretic Measurements Samples containing 1.0 ml. of medium 199, 0-1 ml. of antigen in medium 199, 1-0 ml. of the lymphocyte preparation, and 1.0 ml. of the macrophage suspension were incubated at room temperature for at least ninety minutes before the electrophoretic mobility of the macrophages was measured in a Zeiss ’Cytopherometer’. A duplicate suspension without E.F. was prepared as a control from each sample. Samples were scrambled and measured in a random order, except when scrambling was precluded by other investigations proceeding on the same sample. Cells in focus in the stationary layer were timed over one division of the eyepiece graticule (16 m.) for both directions of current, and the pairs of times recorded if they did not differ by more than 10%. Results were tabulated in two columns, one containing all values close to the control time and the other containing times appreciably longer than the control. Measurements on a sample were stopped as soon as either column contained 10 pairs of timings, and these were averaged to give the mean migration time for
* Methylcellulose technique used (otherwise ficol-triosil). t Lymphocytes stored overnight at 4°C. t Lymphocytes stored for 72 hours at 4°C.
§ Measurements in modified WHS-designed apparatus. s.C.C. Squamous-cell carcinoma. B.C.C. = Basal-cell carcinoma. =
629 TABLE III-PERCENTAGE SLOWING OF MACROPHAGES FROM NORMAL CONTROLS AGAINST E.F. ANTIGEN
healthy hospital staff. We have now extended the investigation to include controls with various nonmalignant illnesses. We have not detected the sensitisation of laboratory staff to E.F., which was an important feature of the normal results quoted by Field and Caspary,l perhaps because we have not yet had sufficient exposure to E.F. For most of the measurements in tables 11 and III, between 1 x 106 and 5 x 106 lymphocytes per ml. were used. Except for the first 3 samples, these were prepared by the ficol-triosil technique, which always produced very good yields with minimum contamination by polymorphs and red blood-cells, and required only sixty minutes of preparation time per sample in contrast to the hundred and thirty minutes required by the methylcellulose technique used by Field and Caspary. The percentage slowing is certainly dependent on lymphocyte numbers, but above 106 lymphocytes per ml. the effect approaches a maximum. We are now attempting, already with some success, to simplify and improve the M.E.M. test as a clinical technique and an important tool in experimental cancer research.
t, § As in table
II.
Subject 54 had a history of sarcoidosis.
ity (M.E.M.) test. In the first 230 samples some slowing was observed in 70% of samples from subjects with malignant disease, but we did not immediately achieve the consistent results claimed by Field and Caspary. After a detailed re-evaluation of all steps in our experimental procedures, we now know that during the period that these 230 samples were tested several adverse factors were operating. These factors will be described elsewhere, together with a full account of our experimental and measurement techniques and an analysis of the frequency and possible significance of the wrong pairs of timings. After the removal of these adverse factors in May, 1972, 79 consecutive samples were measured, 52 from cases of known malignant disease (table 11) and 27 from non-malignant controls (table ill). The results from these 79 samples are shown in tables 11 and ill together with age, sex, and diagnosis. Samples from a few subjects were measured a second time; this is indicated by the figures in parentheses beside the subject number.
We thank our radiotherapist colleagues for their advice and discussion; especially Prof. E. J. Field and E. A. Caspary of the Medical Research Council Unit for Demyelinating Diseases, Newcastle upon Tyne; and the staff of the Tenovus Laboratory and the animal house. This project was financed by Tenovus, the South Wales and Monmouthshire Cancer Research Council, and the Department of Health and Social Security. Requests for reprints should be addressed to J. A. V. P. REFERENCES
1. 2. 3. 4. 5.
Field, E. J., Caspary, E. A. Lancet, 1970, ii, 1337. Caspary, E. A., Field, E. J. Ann. N.Y. Acad. Sci. 1965, 122, 182. Hughes, D., Caspary, E. A. Int. Archs Allergy appl. Immun. 1970, 37, 506. Caspary, E. A., Field, E. J. Br. med. J. 1971, ii, 613. Caspary, E. A., Field, E. J. ibid. p. 143.
SECOND MALIGNANT CLONE UNDERLYING A BURKITT-TUMOUR
EXACERBATION
P. J. FIALKOW Departments of Medicine and Genetics, University of Washington, Seattle, Washington, G. KLEIN
Discussion
We confirmed the results of Field and Caspary 1,4 and our results underline the potential of the M.E.M. test for indicating the presence of malignant disease. Lymphocytes from patients with early or advanced disease were equally effective in producing M.S.F. Tables u and ill confirm the unexpectedly wide gap between malignant and normal samples reported by Field and Caspary. In our series, all samples from patients with known malignant conditions gave macrophage slowing greater than 13%, while all control subjects were below 3-5%, with one exception. A sample from subject 54, who was thought to be normal, gave an unusually high result in an initial and a repeat test. Further investigation revealed a history of sarcoidosis, a condition which Caspary and Field reported as giving a positive result in the .B!.LB1. test. The remainder of our controls were
U.S.A.
Department of Tumour Biology, Karolinska Institutet, Stockholm, Sweden P. CLIFFORD *
Department of Head and Neck Surgery, Kenyatta National Hospital, Nairobi, Kenya
6-year-old girl with Burkitt lymphoma had a chemotherapeutically induced complete " remission, followed by exacerbation of disease, first in the left parotid gland (a site close to one previously affected by tumour) and then in the left orbit (a previously uninvolved site). Her normal tissues were typed as A—B for glucose-6phosphate dehydrogenase (G.-6-P.D.). An ovarian tumour studied before therapy and a recurrentA
Summary
"
*
Present address:
Royal Marsden Hospital, London S.W.3.
the severity and duration of adverse circumstances. The long-term effects of such experiences depend to what extent the harmony of growth 11 is disturbed. My data on obese children show that overnutrition in the early months of life causes not only an increase in linear growth, but also an equivalent advance in skeletal maturation.12 Thus the height of obese children is normal for their bone-age, and the prognosis for height is also normal." Overnutrition later in childhood has no demonstrable effects on height The adipose organ is only or skeletal maturation.12 affected by overnutrition in the early period, but the increase in the number of cells in this event is much greater than the increase in height or skeletal maturity; thus the harmony is destroyed and these individuals will have a permanently increased complement of adipose cells. Since these findings do not apply to obesity dating from later childhood, it seems likely that the first year of life is a sensitive one for the formation of adipose cells. During such a period, cell replication is sensitive to external influences which may be beneficial (perhaps essential) or pathological. The period is also one of determination, because at its end the basic complement of cells is established and thereafter cell replication probably continues at a normal rate regardless of other circumstances. As far as the adipose organ is concerned, it seems that after the end of the sensitive period alterations in nutrition affect only cell size and not cell number. From studies in animals of experimental overnutrition 14 and undernutrition, 15 it is known that during certain periods it is more easy to impair growth and that, in general, the earlier the stress, the more This work has been confirmed severe the effects. 16 and extended by the demonstration of the importance of early nutritional experiences on the size, number, and rate of growth of adipose cells in rats.’-’,18 The effects of intrauterine malnutrition (L.F.D. children) in the present study demonstrate how adverse circumstances early in organogenesis have profound effects later, and how important timing is in growth and development. If G.H. is important for adipose-cell replication (as the present data indicate), an intermediate situation between controls and L.F.D. would be predicted for children with isolated G.H. deficiency. Such children grow normally in utero and are presumably born with a normal complement of cells. These cells do not receive a proper stimulus for replication in the sensitive period and so they lag behind the rest of development as measured by skeletal maturity, but not as much as they would have done if the stress had occurred earlier. Children who lack G.H. at a later age, however, would be expected to have passed the age when the adipose organ is
sensitive, and so harmony should not have been destroyed. Such children would be expected to have a number of adipose cells which does not differ from normal when the delay in growth induced by G.H. deficiency, and evidenced by skeletal retardation, is allowed for. The findings on the C.R. children confirm this expectation. I suggest that the adipose organ in man has a finite and well-defined sensitive period during which external
circumstances have long-term effects on cellular growth. For this organ the period of determination, when the basic complement of cells is established,
probably extends from thirty weeks of intrauterine growth through to the first nine months to a year of extrauterine growth when the skinfolds cease their rapid increase. Presumably such a model, with different timings, applies to all organs in man. I thank the Wellcome Trust and the Medical Research Council for financial assistance, and Prof. J. M. Tanner, whose unfailing advice and support have been of great help. REFERENCES 1. Winick, M. Med. Clins N. Am. 1970, 54, 1413. 2. Cheek, D. B., Hill, D. E., Cordano, A., Graham, G. G. Pediat. Res. 1970, 4, 135. 3. Brook, C. G. D. Archs Dis. Childh. 1971, 46, 182. 4. Tanner, J. M., Whitehouse, R. H. Br. med. J. 1962, i, 446. 5. Brook, C. G. D., Lloyd, J. K., Wolff, O. H. ibid. 1972, ii, 25. 6. Tanner, J. M., Thomson, A. M. Archs Dis. Childh. 1970, 45, 566. 7. Edwards, M. W., Hammond, W. H., Healy, M. J. R., Tanner, J. M., Whitehouse, R. H. Br. J. Nutr. 1955, 9, 133. 8. Durmin, J. V. G. A., Rahaman, M. M. ibid. 1967, 21, 681. 9. Tanner, J. M., Whitehouse, R. H., Healy, M. J. R. A New System for Estimating Skeletal Maturity from the Hand and Wrist. Centre Internationale de L’Enfance, Paris, 1962. 10. Hirsch, J., Gallian, E. J. Lipid Res. 1968, 9, 110. 11. Widdowson, E. M. Lancet, 1970, i, 901. 12. Brook, C. G. D. M.D. thesis, University of Cambridge, 1972. 13. Lloyd, J. K., Wolff, O. H., Whelen, W. S. Br. med. J. 1961, ii, 145. 14. Widdowson, E. M., McCance, R. A. Proc. R. Soc. 1960, B, 152, 188. 15. Widdowson, E. M., McCance, R. A. ibid. 1963, B, 158, 329. 16. Winick, M., Rosso, P. Pediat. Res. 1969, 3, 181. 17. Knittle, J. L., Hirsch, J. J. clin. Invest. 1968, 47, 2091. 18. Hirsch, J., Han, P. W. J. Lipid Res. 1969, 10, 77.
MACROPHAGE-ELECTROPHORETICMOBILITY (M.E.M.) TEST FOR MALIGNANT DISEASE An
Independent Confirmation
J. A. V. PRITCHARD W. H. SUTHERLAND
J. L. MOORE C. A. F. JOSLIN
Tenovus Laboratories, Velindre
Hospital,
Whitchurch, Cardiff
Sum ary
The test
validity of a new in-vitro bloodfor cancer, which depends on the
sensitisation of the patient’s lymphocytes to a common antigen apparently present in human tumours, has been independently confirmed. Introduction
Field and Casparyreported that lymphocytes from patients with malignant disease can be stimulated by encephalitogenic factor2 (E.F.) (a basic protein derived from the human brain) to release a macrophage-slowing factor (M.S.F.) which reduces the electrophoretic mobility of guineapig macrophages. With few exceptions, the effect was not found with lymphocytes from non-malignant controls, and therefore the technique was presented as a possible in-vitro blood-test for cancer. The results could best be explained on the basis of a common antigen present in human tumours. These unexpected findings required confirmation, and we performed a similar investigation. Materials and Methods Approximately 15 ml. of venous blood was collected from healthy hospital staff and from patients with known
628 TABLE
I-TYPICAL
RECORD
OF
TIMINGS
FROM
TWO
SAMPLES
that sample. Typical timings from two samples, one " control " and one " malignant ", are shown in table I. The frequency of " wrong " pairs decreased with increasing experience of the technique and apparatus, until only about 1 cell in every 100 from control samples gave a " wrong " pair of timings. Results were expressed as a percentage change in mobility, 100 (T—c)/c, where T is the average migration time for macrophages in the test sample containing lymphocytes, macrophages, and antigen, and c is the average migration time for macrophages in the control sample containing lymphocytes and macrophages but no
antigen. Results
Between May, 1971, and July, 1972, 309 bloodsamples from patients and from healthy subjects were examined with the macrophage-electrophoretic-mobilTABLE II-PERCENTAGE SLOWING OF MACROPHAGES FROM
SUBJECTS
WITH MALIGNANT DISEASE AGAINST E.F. ANTIGEN
*
This represents
a
t This represents
a
2-7% decrease in mean migration time. 20-6% increase in mean migration time.
After defibrination, lymphocytes were harvested either by the methylcellulose technique described by Hughes and Caspary3 or by means of a ’FicolTriosil ’ technique which will be described fully elsewhere. The washed lymphocytes from either technique were Macrophages were resuspended in’Medium 199’. prepared from the peritoneal exudate of 400-600 g. Hartley albino guineapigs four to ten days after the injection of 20 ml. of sterile liquid paraffin into the peritoneum. The cells were washed and resuspended in medium 199 to a concentration of 10’ cells per ml. and then irradiated with a dose of 100-200 rad from a cxsium-137 source. The encephalitogenic factor (E.F.) was prepared from a human brain by the methods described by Caspary and Field.2,4 1-0 mg. of E.F. was dissolved in medium 199 so that the final concentration was 100 (kg. per 0-1 ml.
malignant disease.
Electrophoretic Measurements Samples containing 1.0 ml. of medium 199, 0-1 ml. of antigen in medium 199, 1-0 ml. of the lymphocyte preparation, and 1.0 ml. of the macrophage suspension were incubated at room temperature for at least ninety minutes before the electrophoretic mobility of the macrophages was measured in a Zeiss ’Cytopherometer’. A duplicate suspension without E.F. was prepared as a control from each sample. Samples were scrambled and measured in a random order, except when scrambling was precluded by other investigations proceeding on the same sample. Cells in focus in the stationary layer were timed over one division of the eyepiece graticule (16 m.) for both directions of current, and the pairs of times recorded if they did not differ by more than 10%. Results were tabulated in two columns, one containing all values close to the control time and the other containing times appreciably longer than the control. Measurements on a sample were stopped as soon as either column contained 10 pairs of timings, and these were averaged to give the mean migration time for
* Methylcellulose technique used (otherwise ficol-triosil). t Lymphocytes stored overnight at 4°C. t Lymphocytes stored for 72 hours at 4°C.
§ Measurements in modified WHS-designed apparatus. s.C.C. Squamous-cell carcinoma. B.C.C. = Basal-cell carcinoma. =
629 TABLE III-PERCENTAGE SLOWING OF MACROPHAGES FROM NORMAL CONTROLS AGAINST E.F. ANTIGEN
healthy hospital staff. We have now extended the investigation to include controls with various nonmalignant illnesses. We have not detected the sensitisation of laboratory staff to E.F., which was an important feature of the normal results quoted by Field and Caspary,l perhaps because we have not yet had sufficient exposure to E.F. For most of the measurements in tables 11 and III, between 1 x 106 and 5 x 106 lymphocytes per ml. were used. Except for the first 3 samples, these were prepared by the ficol-triosil technique, which always produced very good yields with minimum contamination by polymorphs and red blood-cells, and required only sixty minutes of preparation time per sample in contrast to the hundred and thirty minutes required by the methylcellulose technique used by Field and Caspary. The percentage slowing is certainly dependent on lymphocyte numbers, but above 106 lymphocytes per ml. the effect approaches a maximum. We are now attempting, already with some success, to simplify and improve the M.E.M. test as a clinical technique and an important tool in experimental cancer research.
t, § As in table
II.
Subject 54 had a history of sarcoidosis.
ity (M.E.M.) test. In the first 230 samples some slowing was observed in 70% of samples from subjects with malignant disease, but we did not immediately achieve the consistent results claimed by Field and Caspary. After a detailed re-evaluation of all steps in our experimental procedures, we now know that during the period that these 230 samples were tested several adverse factors were operating. These factors will be described elsewhere, together with a full account of our experimental and measurement techniques and an analysis of the frequency and possible significance of the wrong pairs of timings. After the removal of these adverse factors in May, 1972, 79 consecutive samples were measured, 52 from cases of known malignant disease (table 11) and 27 from non-malignant controls (table ill). The results from these 79 samples are shown in tables 11 and ill together with age, sex, and diagnosis. Samples from a few subjects were measured a second time; this is indicated by the figures in parentheses beside the subject number.
We thank our radiotherapist colleagues for their advice and discussion; especially Prof. E. J. Field and E. A. Caspary of the Medical Research Council Unit for Demyelinating Diseases, Newcastle upon Tyne; and the staff of the Tenovus Laboratory and the animal house. This project was financed by Tenovus, the South Wales and Monmouthshire Cancer Research Council, and the Department of Health and Social Security. Requests for reprints should be addressed to J. A. V. P. REFERENCES
1. 2. 3. 4. 5.
Field, E. J., Caspary, E. A. Lancet, 1970, ii, 1337. Caspary, E. A., Field, E. J. Ann. N.Y. Acad. Sci. 1965, 122, 182. Hughes, D., Caspary, E. A. Int. Archs Allergy appl. Immun. 1970, 37, 506. Caspary, E. A., Field, E. J. Br. med. J. 1971, ii, 613. Caspary, E. A., Field, E. J. ibid. p. 143.
SECOND MALIGNANT CLONE UNDERLYING A BURKITT-TUMOUR
EXACERBATION
P. J. FIALKOW Departments of Medicine and Genetics, University of Washington, Seattle, Washington, G. KLEIN
Discussion
We confirmed the results of Field and Caspary 1,4 and our results underline the potential of the M.E.M. test for indicating the presence of malignant disease. Lymphocytes from patients with early or advanced disease were equally effective in producing M.S.F. Tables u and ill confirm the unexpectedly wide gap between malignant and normal samples reported by Field and Caspary. In our series, all samples from patients with known malignant conditions gave macrophage slowing greater than 13%, while all control subjects were below 3-5%, with one exception. A sample from subject 54, who was thought to be normal, gave an unusually high result in an initial and a repeat test. Further investigation revealed a history of sarcoidosis, a condition which Caspary and Field reported as giving a positive result in the .B!.LB1. test. The remainder of our controls were
U.S.A.
Department of Tumour Biology, Karolinska Institutet, Stockholm, Sweden P. CLIFFORD *
Department of Head and Neck Surgery, Kenyatta National Hospital, Nairobi, Kenya
6-year-old girl with Burkitt lymphoma had a chemotherapeutically induced complete " remission, followed by exacerbation of disease, first in the left parotid gland (a site close to one previously affected by tumour) and then in the left orbit (a previously uninvolved site). Her normal tissues were typed as A—B for glucose-6phosphate dehydrogenase (G.-6-P.D.). An ovarian tumour studied before therapy and a recurrentA
Summary
"
*
Present address:
Royal Marsden Hospital, London S.W.3.