880 various circulatory diseases fails to support the hypothesis that smoking causes these diseases. If these various findings and arguments are valid we have .to conclude that the risk of death from various circulatory diseases is raised by the use of oral contraceptives-at least among those women who are genetically predisposed to such disorders-but that smoking does not increase the risk. cause
General Infirmary, Leeds LS1 3EX
P. R.
J. BURCH
COMMON CLONAL ORIGIN OF LYMPHOPLASMACYTIC PROLIFERATION AND IMMUNOBLASTIC LYMPHOMA IN INTESTINAL &agr;-CHAIN DISEASE
SIR,-The development of immunoblastic lymphoma in the intestinal form of a-chain disease (a-C.D.) has been morphologically confirmed.’-6 There is also evidence for a common clonal origin for the lymphoplasmacytic proliferation and immunoblastic lymphoma associated with it in two cases of a-C.D.7.8 We have searched for intracellular immunoglobulin in tissue sections from both the benign-appearing lymphoplasmacytic lesions and the immunoblastic lymphomas in three cases with &agr;—C.D., using the peroxidase/antiperoxidase reaction,9,10 as follows : (1) Sections deparaffinised and hydrated with water. (2) Endogenous peroxidase activity blocked with fresh 3% solution of hydrogen peroxide in "tris"-saline buffer for 30 min. (3) Non-specific background staining reduced with normal nonimmune non-conjugated goat serum diluted 1/10 for 10 min. Excess serum removed but not washed off before next stage. (4) Sections treated with monospecific rabbit antisera for 30 min in the following dilutions: 1/40 for anti a, y, and µ, and 1/100 for anti x and &lgr;. All antisera were obtained from Dakopatts A/S Denmark, 1.
Rambaud, J. C., Bognel, C., Prost, A., Bernier, J. J., LeQuintrec, Y., Lambling, A., Danon, F., Hurez, D., Seligmann, M. Digestion, 1968, 1, 321. 2. Rappaport, H., Ramot, B., Hulu, N., Park, J. K. Cancer, 1972, 29, 1502. 3. Rappaport, H. in Maligne Lymphome und Monoklonale Gammophathien (edited by H. Löffler); p. 377. Munich, 1976. 4. Tabbane, S., Tabbane, F., Cammoun, M., Mourall, N. Cancer, 1976, 38, 1989. 5.
Galian, A., Lecestre, M.-J., Scotto, J., Bognel, C., Matuchansky, C., Rambaud, J.-C. ibid. 1977, 39, 2081. 6. Rappaport, H., Pangalis, G. A., Nathwani, B. N. Cold Spring Harbor Symp.
(in the press). 7. Brouet, J. C., Mason, D. Y., Danon, F.,
Preud’homme, J. L., Seligmann, M., Reyes, F., Navab, F., Galian, A., Rene, E., Rambaud, J. C. Lancet, 1977, i, 861. 8. Ramot, B., Levanon, M., Hahn, Y., Lahat, N., Moroz, C. Clin. exp. Immun. 1977, 27, 440. 9. Sternberger, L. A., Hardy, P. H., Jr., Cuculis, J. J., Meyer, H. G. J. Histochem. Cytochem. 1970, 18, 315. 10. Taylor, C. R. Eur. J. Cancer, 1976, 12, 61.
Fig. 2-Immunoblastic lymphoma
in x-C.D.:
immunoperoai-
dase stain.
Many tumour cells positive for a-chains (x 270).
through the Accurate Chemical and Scientific Corporation, Hicksville, N.Y.
(5) Unlabelled
swine anti-rabbit
serum
protein diluted 1/20 for 30
min.
(6) Rabbit peroxidase/antiperoxidase complex diluted 1/100 for 30 min.
(7) Diaminobenzidine reaction
4 min. Wash in tris-saline for 30
min.
(8) Sections counterstained with hæmatoxylin (4 min), eosin (1 s), dehydrated, cleared in xylol and mounted in synthetic medium. The reactions were done in 50 mmol/1 "tris" buffer (pH 7.6) diluted 1/10 with saline. "Tris"-saline was also used for diluting the sera and the washings for 30 min after stages 2, 4, 5, and 6. In all cases a high percentage of plasma cells and plasmacytoid lymphocytes of the morphologically benign-appearing cellular proliferation were specifically and strongly positive for a-chains (fig. 1), while adjacent sections were entirely negative for &ggr;, µ, x and X chains. A similar positivity for a-chains was observed in the neoplastic cells of the immunoblastic lymphomas (fig. 2), while antisera to Y, µ, x and a chains in adjacent sections gave negative results. Our findings further support the concept of the common clonal origin of the lymphoplasmacytic proliferation and the immunoblastic lymphoma in intestinal cx-C.D. In addition, the technique described could be an excellent diagnostic tool for the recognition of a-c.D. in biopsy material, particularly since the identification of the abnormal a chain in serum is sometimes difficult. Department of Anatomic Pathology, City of Hope National Medical Center, Duarte, California 91010, U.S.A.
G. A. PANGALIS H. RAPPAPORT
TRISOMY CLUSTER IN NEW YORK
SIR,—Dr
Warburton and her colleagues (July 23, p. 201) for the cluster of trisomic karyotypes in conceptions that had taken place in the winter of 1976 in New York. Perhaps examination of this finding was restricted totime and place-rather than-widened to the universal phenomenon of seasonality of birth (or of conception) of patients with chromosomal aberrations, congenital malformations, or psychopathology. The months of last menstrual period (L.M.P.) associated with a high rate of trisomy conceptions were also those with a high rate of conceptions leading to spontaneous abortion (i.e., without chromosomal aberrations). In the summer of 1966 an apparent surge in the birth-rate of New York was linked by the lay Press throughout the world with the electricity blackout nine months before. However, Udryl demonstrated that there was no significant difference could find
Fig. 1—&agr;-C.D.: immunoperoxidase stain. Many cells positive for x-chains (arrows) (x270).
1.
no cause
Udry, J. R. Demography, 1970, 7, 325.
General Infirmary, Leeds LS1 3EX
P. R.
J. BURCH
COMMON CLONAL ORIGIN OF LYMPHOPLASMACYTIC PROLIFERATION AND IMMUNOBLASTIC LYMPHOMA IN INTESTINAL &agr;-CHAIN DISEASE
SIR,-The development of immunoblastic lymphoma in the intestinal form of a-chain disease (a-C.D.) has been morphologically confirmed.’-6 There is also evidence for a common clonal origin for the lymphoplasmacytic proliferation and immunoblastic lymphoma associated with it in two cases of a-C.D.7.8 We have searched for intracellular immunoglobulin in tissue sections from both the benign-appearing lymphoplasmacytic lesions and the immunoblastic lymphomas in three cases with &agr;—C.D., using the peroxidase/antiperoxidase reaction,9,10 as follows : (1) Sections deparaffinised and hydrated with water. (2) Endogenous peroxidase activity blocked with fresh 3% solution of hydrogen peroxide in "tris"-saline buffer for 30 min. (3) Non-specific background staining reduced with normal nonimmune non-conjugated goat serum diluted 1/10 for 10 min. Excess serum removed but not washed off before next stage. (4) Sections treated with monospecific rabbit antisera for 30 min in the following dilutions: 1/40 for anti a, y, and µ, and 1/100 for anti x and &lgr;. All antisera were obtained from Dakopatts A/S Denmark, 1.
Rambaud, J. C., Bognel, C., Prost, A., Bernier, J. J., LeQuintrec, Y., Lambling, A., Danon, F., Hurez, D., Seligmann, M. Digestion, 1968, 1, 321. 2. Rappaport, H., Ramot, B., Hulu, N., Park, J. K. Cancer, 1972, 29, 1502. 3. Rappaport, H. in Maligne Lymphome und Monoklonale Gammophathien (edited by H. Löffler); p. 377. Munich, 1976. 4. Tabbane, S., Tabbane, F., Cammoun, M., Mourall, N. Cancer, 1976, 38, 1989. 5.
Galian, A., Lecestre, M.-J., Scotto, J., Bognel, C., Matuchansky, C., Rambaud, J.-C. ibid. 1977, 39, 2081. 6. Rappaport, H., Pangalis, G. A., Nathwani, B. N. Cold Spring Harbor Symp.
(in the press). 7. Brouet, J. C., Mason, D. Y., Danon, F.,
Preud’homme, J. L., Seligmann, M., Reyes, F., Navab, F., Galian, A., Rene, E., Rambaud, J. C. Lancet, 1977, i, 861. 8. Ramot, B., Levanon, M., Hahn, Y., Lahat, N., Moroz, C. Clin. exp. Immun. 1977, 27, 440. 9. Sternberger, L. A., Hardy, P. H., Jr., Cuculis, J. J., Meyer, H. G. J. Histochem. Cytochem. 1970, 18, 315. 10. Taylor, C. R. Eur. J. Cancer, 1976, 12, 61.
Fig. 2-Immunoblastic lymphoma
in x-C.D.:
immunoperoai-
dase stain.
Many tumour cells positive for a-chains (x 270).
through the Accurate Chemical and Scientific Corporation, Hicksville, N.Y.
(5) Unlabelled
swine anti-rabbit
serum
protein diluted 1/20 for 30
min.
(6) Rabbit peroxidase/antiperoxidase complex diluted 1/100 for 30 min.
(7) Diaminobenzidine reaction
4 min. Wash in tris-saline for 30
min.
(8) Sections counterstained with hæmatoxylin (4 min), eosin (1 s), dehydrated, cleared in xylol and mounted in synthetic medium. The reactions were done in 50 mmol/1 "tris" buffer (pH 7.6) diluted 1/10 with saline. "Tris"-saline was also used for diluting the sera and the washings for 30 min after stages 2, 4, 5, and 6. In all cases a high percentage of plasma cells and plasmacytoid lymphocytes of the morphologically benign-appearing cellular proliferation were specifically and strongly positive for a-chains (fig. 1), while adjacent sections were entirely negative for &ggr;, µ, x and X chains. A similar positivity for a-chains was observed in the neoplastic cells of the immunoblastic lymphomas (fig. 2), while antisera to Y, µ, x and a chains in adjacent sections gave negative results. Our findings further support the concept of the common clonal origin of the lymphoplasmacytic proliferation and the immunoblastic lymphoma in intestinal cx-C.D. In addition, the technique described could be an excellent diagnostic tool for the recognition of a-c.D. in biopsy material, particularly since the identification of the abnormal a chain in serum is sometimes difficult. Department of Anatomic Pathology, City of Hope National Medical Center, Duarte, California 91010, U.S.A.
G. A. PANGALIS H. RAPPAPORT
TRISOMY CLUSTER IN NEW YORK
SIR,—Dr
Warburton and her colleagues (July 23, p. 201) for the cluster of trisomic karyotypes in conceptions that had taken place in the winter of 1976 in New York. Perhaps examination of this finding was restricted totime and place-rather than-widened to the universal phenomenon of seasonality of birth (or of conception) of patients with chromosomal aberrations, congenital malformations, or psychopathology. The months of last menstrual period (L.M.P.) associated with a high rate of trisomy conceptions were also those with a high rate of conceptions leading to spontaneous abortion (i.e., without chromosomal aberrations). In the summer of 1966 an apparent surge in the birth-rate of New York was linked by the lay Press throughout the world with the electricity blackout nine months before. However, Udryl demonstrated that there was no significant difference could find
Fig. 1—&agr;-C.D.: immunoperoxidase stain. Many cells positive for x-chains (arrows) (x270).
1.
no cause
Udry, J. R. Demography, 1970, 7, 325.
