204
Letters
to
the Editor
ALLOANTIBODIES REACTING WITH HUSBAND’S B-LYMPHOCYTES IN SERA FROM WOMEN WITH TROPHOBLASTIC NEOPLASIA
SIR,-Trophoblastic neoplasms can be considered as maligsemiallogeneic grafts. Alloimmunisation against the husband’s HLA antigens has been shown in patients with placental choriocarcinoma. Genes coding for HLA antigens are closely linked to genes coding for alloantigens restricted to B nant
ERYTHROCYTE SUPEROXIDE-DISMUTASE DEFICIENCY IN FANCONI’S ANÆMIA
SiR,-Characteristic symptoms of constitutional infantile
lymphocytes.2.3 We have determined whether sera from women
panmyelopathy (Fanconi’s anaemia) include short stature, microcephaly, hyperpigmentation of the skin, progressive bonemarrow insufficiency, multiple skeletal malformations, and kidney malformations; a high frequency of chromosomal aberrations (e.g., gaps, breaks, and fragments) is typical.’ This "spontaneous" chromosomal damage has been attributed to defective D.N.A. repair.2-5 However, an alternative possibility
with trophoblastic neoplasia contain alloantibodies to the B cells of the donor’s husband. 16 women were studied before any chemotherapy. In twostep microlymphocytotoxicity assays4 13 sera reacted with
should be considered.
Oxygen-metabolising cells generate the superoxide free radical, 02-, which, with its reaction products, hydroxyl radicals and singlet-excited oxygen, could provide a continuous background of chromosome-damaging agents to which all aerobically living cells are exposed.6 Cells are protected against these agents by superoxide dismutase, an enzyme which specifically quenches 0." A deficiency of this enzyme might lead to increased cellular concentrations of 02-, so that the cell’s D.N.A. repair capacity cannot cope and chromosome damage might ensue. Thus, a deficiency of superoxide dismutase might be an alternative explanation for the increased risk of chromosomal anomalies borne by the patient with Fanconi’s anaemia. Nordenson’s observation that Fanconi’s anaemia cells show less chromosomal aberrations when cultured in the presence of added superoxide dismutase and/or catalase points in this direction. We have obtained support for the above hypothesis by measuring erythrocyte superoxide dismutase in two unrelated patients with typical Fanconi’s anaemia. Patient A was a 19-yearold male with stunted growth, pancytopenia, cafe-au-lait spots, hypogenitalism, a horseshoe kidney, and a high incidence of chromosomal breaks in cultured lymphocytes. The second patient (B) was a 32-year-old female with short stature with microcephaly, skeletal malformations (thumbs absent), strabismus, pancytopenia, cafe-au-lait spots, deafness, renal anomalies ; lymphocytes, which grew poorly in culture, showed many chromosomal aberrations. Erythrocyte superoxide dismutase activity was determined by the xanthine-oxidase method of Beauchamp and Fridovich8 and by Mancini’s immunodiffusion technique. With both techniques erythrocyte superoxide dismutase was markedly reduced in the two patients. In patient A a reduction of 26% (activity) or 30% (immunodiffusion) was found; in patient B values were lowered by 39% and 33%, respectively. Since the standard deviation of the individual normal activity values is 8.2%, these reductions, more than 3 and almost 5 S.D.S below the normal mean, support the hypothesis that the enhanced spontaneous chromosomal damage in Fanconi’s anaemia could be due to decreased levels of cellular superoxide dismutase.
Institute of Human Genetics, Free University, 1011 Amsterdam, Netherlands
H. JOENJE A. W. ERIKSSON R. R. FRANTS F. ARWERT
Department of Hæmatology, University Hospital, Groningen
B. HOUWEN
1.
Schroeder,
T.
15-20% unfractionated peripheral blood lymphocytes from the donor’s husband. 3 sera reacted with 80-100% cells and were shown to contain anti-HLA antibodies. The titres of these 16 sera ranged from 1 :64 to 1 :512. Similar results were obtained after extensive absorption of each serum with packed pooled human platelets (which carry HLA antigens, but not B-lymphocyte alloantigens). After absorption the 3 sera with anti-HLA antibodies killed only
blood lymphocytes.
Lymphocyte subpopulations enriched in or depleted of T were obtained by centrifugation of sheep-erythrocyte (E)rosette-forming cells through ’Ficoll-Hypaque’. Purified T cells (99% E-rosette-forming cells) failed to react with the patients’ sera but 50-70% purified B cells were killed. 3 sera were studied by indirect immunofluorescence. Alloantibodies reacting with lymphocytes from the donor’s husband could be stained at the cell surface with rhodamine-conjugated F(ab’)2 anti-Y fragments (kindly provided by Dr J. L. Preud’homme), and thus were shown to belong to the IgG class. The reactivity of each serum with B lymphocytes from a panel of normal subjects showed that the target antigens were cells
Preliminary
data are consistent restricted to some individuals. with an allelic distribution. These results strongly suggest that each of the 16 sera contained alloantibodies, clearly distinct from anti-HLA antibodies, directed against B lymphocytes from the donor’s husband. Alloantibodies to B lymphocytes are detectable in sera from multiparous women and in about 3% of pregnancy sera.s The antibodies may appear in a larger proportion of women with trophoblastic malignancies, since we detected them in all the 16 sera tested. 5 patients were primiparous, ruling out immunisation by previous pregnancies as a cause. The titre of these antibodies was unusually high, since in pregnancy sera it seldom exceeds 1:32. Though consistently present at the time of diagnosis, antibodies to B lymphocytes were not detectable in 6 out of 12 sera from patients treated (and apparently
cured) by chemotherapy. Laboratoire d’Immuno-hématologie et Service de Médecine "A", Institut Gustave-Roussy, 94800 Villejuif, France
THOMAS TURSZ MARYVONNE GUILLARD MARC LIPINSKI
J. L. AMIEL
CORONARY HEART-DISEASE AFTER TREATMENT OF HYPERTENSION
SiR,—Dr Berglund and his colleagues (Jan. 7, p. 1) have achieved
a small drop in the rate of myocardial infarction in middle-aged moderate hypertensives under treatment compared with controls. They believe that the reason for the failure of earlier attempts to protect rather severe hypertensives
M., Tilgen, D., Krüger, J., Vogel, F. Hum. Genet. 1976, 32,
257.
1.
Sasaki, M. S., Tonomura, A. Cancer Res. 1973, 33, 1829. 3. Poon, P. K., O’Brien, R. L., Parker, J. W. Nature, 1974, 250, 223. 4. Latt, S. A., Stetten, G., Juergens, L. A., Buchanan, G. R., Gerald, P. S. Proc. natn. Acad. Sci. U.S.A. 1975, 72, 4066. 5. Sasaki, M. S. Nature, 1975, 257, 501. 2.
6. Fridovich, I. Ann. Rev. Biochem. 1975, 44, 147. 7. Nordenson, I. Hereditas, 1977, 86, 147. 8. Beauchamp, C., Fridovich, I. Analyt. Biochem. 1971,
44, 276.
2.
3. 4. 5.
Mathé, G., Dausset, J., Hervet, E., Amiel, J. L., Colombani, J. J. nat. Cancer Inst. 1974, 33, 193. van Rood, J. J., van Leeuwen, A., Parlevliet, J., Termijtelen, A., Keuning, J. J. in Histocompatibility Testing 1975 (edited by F. Kissmeyer-Nielsen); p. 629. Copenhagen, 1976. Bodmer, W. F., Jones, E. A., Young, D., Goodfellow, P. N., Bodmer, J. G., Dick, H. M., Steel, C. M. ibid. p. 677. Terasaki, P. I., MacClelland, J. D. Nature, 1964, 206, 998. Wernet, P. Transpl. Rev. 1976, 30, 270.
Letters
to
the Editor
ALLOANTIBODIES REACTING WITH HUSBAND’S B-LYMPHOCYTES IN SERA FROM WOMEN WITH TROPHOBLASTIC NEOPLASIA
SIR,-Trophoblastic neoplasms can be considered as maligsemiallogeneic grafts. Alloimmunisation against the husband’s HLA antigens has been shown in patients with placental choriocarcinoma. Genes coding for HLA antigens are closely linked to genes coding for alloantigens restricted to B nant
ERYTHROCYTE SUPEROXIDE-DISMUTASE DEFICIENCY IN FANCONI’S ANÆMIA
SiR,-Characteristic symptoms of constitutional infantile
lymphocytes.2.3 We have determined whether sera from women
panmyelopathy (Fanconi’s anaemia) include short stature, microcephaly, hyperpigmentation of the skin, progressive bonemarrow insufficiency, multiple skeletal malformations, and kidney malformations; a high frequency of chromosomal aberrations (e.g., gaps, breaks, and fragments) is typical.’ This "spontaneous" chromosomal damage has been attributed to defective D.N.A. repair.2-5 However, an alternative possibility
with trophoblastic neoplasia contain alloantibodies to the B cells of the donor’s husband. 16 women were studied before any chemotherapy. In twostep microlymphocytotoxicity assays4 13 sera reacted with
should be considered.
Oxygen-metabolising cells generate the superoxide free radical, 02-, which, with its reaction products, hydroxyl radicals and singlet-excited oxygen, could provide a continuous background of chromosome-damaging agents to which all aerobically living cells are exposed.6 Cells are protected against these agents by superoxide dismutase, an enzyme which specifically quenches 0." A deficiency of this enzyme might lead to increased cellular concentrations of 02-, so that the cell’s D.N.A. repair capacity cannot cope and chromosome damage might ensue. Thus, a deficiency of superoxide dismutase might be an alternative explanation for the increased risk of chromosomal anomalies borne by the patient with Fanconi’s anaemia. Nordenson’s observation that Fanconi’s anaemia cells show less chromosomal aberrations when cultured in the presence of added superoxide dismutase and/or catalase points in this direction. We have obtained support for the above hypothesis by measuring erythrocyte superoxide dismutase in two unrelated patients with typical Fanconi’s anaemia. Patient A was a 19-yearold male with stunted growth, pancytopenia, cafe-au-lait spots, hypogenitalism, a horseshoe kidney, and a high incidence of chromosomal breaks in cultured lymphocytes. The second patient (B) was a 32-year-old female with short stature with microcephaly, skeletal malformations (thumbs absent), strabismus, pancytopenia, cafe-au-lait spots, deafness, renal anomalies ; lymphocytes, which grew poorly in culture, showed many chromosomal aberrations. Erythrocyte superoxide dismutase activity was determined by the xanthine-oxidase method of Beauchamp and Fridovich8 and by Mancini’s immunodiffusion technique. With both techniques erythrocyte superoxide dismutase was markedly reduced in the two patients. In patient A a reduction of 26% (activity) or 30% (immunodiffusion) was found; in patient B values were lowered by 39% and 33%, respectively. Since the standard deviation of the individual normal activity values is 8.2%, these reductions, more than 3 and almost 5 S.D.S below the normal mean, support the hypothesis that the enhanced spontaneous chromosomal damage in Fanconi’s anaemia could be due to decreased levels of cellular superoxide dismutase.
Institute of Human Genetics, Free University, 1011 Amsterdam, Netherlands
H. JOENJE A. W. ERIKSSON R. R. FRANTS F. ARWERT
Department of Hæmatology, University Hospital, Groningen
B. HOUWEN
1.
Schroeder,
T.
15-20% unfractionated peripheral blood lymphocytes from the donor’s husband. 3 sera reacted with 80-100% cells and were shown to contain anti-HLA antibodies. The titres of these 16 sera ranged from 1 :64 to 1 :512. Similar results were obtained after extensive absorption of each serum with packed pooled human platelets (which carry HLA antigens, but not B-lymphocyte alloantigens). After absorption the 3 sera with anti-HLA antibodies killed only
blood lymphocytes.
Lymphocyte subpopulations enriched in or depleted of T were obtained by centrifugation of sheep-erythrocyte (E)rosette-forming cells through ’Ficoll-Hypaque’. Purified T cells (99% E-rosette-forming cells) failed to react with the patients’ sera but 50-70% purified B cells were killed. 3 sera were studied by indirect immunofluorescence. Alloantibodies reacting with lymphocytes from the donor’s husband could be stained at the cell surface with rhodamine-conjugated F(ab’)2 anti-Y fragments (kindly provided by Dr J. L. Preud’homme), and thus were shown to belong to the IgG class. The reactivity of each serum with B lymphocytes from a panel of normal subjects showed that the target antigens were cells
Preliminary
data are consistent restricted to some individuals. with an allelic distribution. These results strongly suggest that each of the 16 sera contained alloantibodies, clearly distinct from anti-HLA antibodies, directed against B lymphocytes from the donor’s husband. Alloantibodies to B lymphocytes are detectable in sera from multiparous women and in about 3% of pregnancy sera.s The antibodies may appear in a larger proportion of women with trophoblastic malignancies, since we detected them in all the 16 sera tested. 5 patients were primiparous, ruling out immunisation by previous pregnancies as a cause. The titre of these antibodies was unusually high, since in pregnancy sera it seldom exceeds 1:32. Though consistently present at the time of diagnosis, antibodies to B lymphocytes were not detectable in 6 out of 12 sera from patients treated (and apparently
cured) by chemotherapy. Laboratoire d’Immuno-hématologie et Service de Médecine "A", Institut Gustave-Roussy, 94800 Villejuif, France
THOMAS TURSZ MARYVONNE GUILLARD MARC LIPINSKI
J. L. AMIEL
CORONARY HEART-DISEASE AFTER TREATMENT OF HYPERTENSION
SiR,—Dr Berglund and his colleagues (Jan. 7, p. 1) have achieved
a small drop in the rate of myocardial infarction in middle-aged moderate hypertensives under treatment compared with controls. They believe that the reason for the failure of earlier attempts to protect rather severe hypertensives
M., Tilgen, D., Krüger, J., Vogel, F. Hum. Genet. 1976, 32,
257.
1.
Sasaki, M. S., Tonomura, A. Cancer Res. 1973, 33, 1829. 3. Poon, P. K., O’Brien, R. L., Parker, J. W. Nature, 1974, 250, 223. 4. Latt, S. A., Stetten, G., Juergens, L. A., Buchanan, G. R., Gerald, P. S. Proc. natn. Acad. Sci. U.S.A. 1975, 72, 4066. 5. Sasaki, M. S. Nature, 1975, 257, 501. 2.
6. Fridovich, I. Ann. Rev. Biochem. 1975, 44, 147. 7. Nordenson, I. Hereditas, 1977, 86, 147. 8. Beauchamp, C., Fridovich, I. Analyt. Biochem. 1971,
44, 276.
2.
3. 4. 5.
Mathé, G., Dausset, J., Hervet, E., Amiel, J. L., Colombani, J. J. nat. Cancer Inst. 1974, 33, 193. van Rood, J. J., van Leeuwen, A., Parlevliet, J., Termijtelen, A., Keuning, J. J. in Histocompatibility Testing 1975 (edited by F. Kissmeyer-Nielsen); p. 629. Copenhagen, 1976. Bodmer, W. F., Jones, E. A., Young, D., Goodfellow, P. N., Bodmer, J. G., Dick, H. M., Steel, C. M. ibid. p. 677. Terasaki, P. I., MacClelland, J. D. Nature, 1964, 206, 998. Wernet, P. Transpl. Rev. 1976, 30, 270.
