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Clinical significance of cytogenetic findings in untreated patients with B-cell chronic lymphocytic leukemia
Clinical Significance of Cytogenetic Findings in Untreated Patients with B-Cell Chronic Lymphocytic Leukemia Naoki Sadamori, Tin Han, Jun Minowada, and Avery A. Sandberg
ABSTRACT: The chromosome constitutions of stimulated lymphocytes in 21 untreated cases with Bcell chronic lymphocytic leukemia (B-CLL) were examined. So-called polyclonal B-cell activators, i.e., pokeweed mitogen, Epstein-Barr virus, and lipopolysaccharide W from E. coil 055:B5, were used. Four out of 21 cases showed abnormal clones with trisomy 12 and were started an therapy shortly after the diagnosis and cytogenetic examination. On the other hand, most cases without abnormal clones had not received treatment for relatively long periods before and after cytogenetic examination. These findings may indicate that cytogenetic results can be utilized as a parameter for treatment and prognosis in B-CLL.
INTRODUCTION Some investigators [1-3] have reported that the most c o m m o n c h r o m o s o m e abnormality of l y m p h o c y t e s stimulated by polyclonal B-cell activators (PBA) in B-cell chronic l y m p h o c y t i c leukemia (B-CLL) was trisomy 12. However, the i n ci d en ce of CLL cases with abnormal c h r o m o s o m e s differs from laboratory to laboratory. Schr6der et al. [4] found an abnormal clone in 1 out of 31 (3.2%) untreated B-CLL cases and in 4 out of 35 (11.4%) treated cases, including those without any metaphases. Gahrton et al. [1] reported that 8 out of 11 (72.7%) untreated and treated cases of B-CLL showed abnormal clones. Morita et al. [2] fou'~d abnormal clones in 9 out of 27 (33.3%) cases, including those w i t h o u t any metaphases. The differences in the i n ci d e n c e of abnormal clones among the B-CLL cases r\eported to date may be related to such factors as stage of disease, interval betwee~ the diagnosis and cytogenetic examination and treatment background. We have already reported that 4 out of 9 (44.4%) untreated and n e w l y diagnosed B-CLL cases [3] and 15 out of 40 (37.5%) treated B-CLL cases showed abnormal clones [5]. In the present study, we attempted to clarify the i n ci d en ce of abnormal clones among untreated cases and the relationship between chromosomal abnormalities and some clinical parameters.
From the Roswell Park Memorial Institute, Buffalo. Address requests for reprints to Avery A. Sandberg, M.D., Roswell Park Memorial Institute, 666 Elm Street, Buffalo, N Y 14263. Received December 1, 1962; accepted March 3, 1983.
45 '~ 1984 by Elsevier Science Publishing Co., Inc. 52 Vanderbilt Ave., New York, NY 10017
Cancer Genetics and Cytogenetics 11, 45 51 (1984) 0165-4608/84/$03.00
46
N. Sadamori et al.
MATERIALS AND METHODS Lymphocytes were obtained from 21 untreated cases with clinically and imnlul~ologically typical B-CLL (14 males and 7 females], whose ages ranged from 49 to 88 years, the mean age being 63 years. The diagnoses were based on accepted clinical and laboratory criteria. Clinical, hematological, and immunological data on the cases are shown in Table 1. No patient in the present study had a history of a¢:cidental or therapentic irradiation or therapy with anticancer drugs. Nine cases leases 1-9) have been already reported hy us 131. The methodologies for E-rosette, EA-rosette, and EAC-rosette formation and surface membrane i m m u n o g l o b u l i n s (SmIg) of the separated lymphocytes have been reported by M i n o w a d a et al. 161. Lymphocytes were separated from 20 ml of heparinized peripheral blood or 2 ml of heparinized bone marrow material by centrifugation over a Ficoll-Hypaque gradient. Separated lymphocytes {1 × 10 '~ cells/roll were cultured for 3 days and harvested according to procedures described by us previously [7, 81. Usually 10-ml cultures were set up: three cultures with pokeweed mitogen (PWM), Epstein-Bari: virus (EBV), and lipopolysaccharide W from E. coli 055:B5 {LPS) were set up t()r each case. Karyotype analyses were performed by G- and Q-banding techniques I,~l. At least 20 optimally banded metaphases were analyzed in each case. An abnormal clone was defined as the presence of two cells with the same extra chromosome or structural rearrangement, or three cells with the same missing chromosome, according to the First International Workshop on Chromosomes in Leukemia [10 I. Treatment {chlorambucil and/or prednisone) was given to all B-CI,i, patients with stage [II-IV disease, as well as to stage [-II patients when their splenomegaly and/or l y m p h a d e n o p a t h y significantly increased. No treatment was usually necessary in patients with stage 0 disease. Case 3 with stage O, associated with monoclonal IgM gammopathy (3,500 mg/ml), was treated with chlorambucil because c)f the d e v e l o p m e n t of peculiar neurological symptoms suggestive of a hyperviscosity syndrome.
RESULTS As shown in Table 2, out of the 21 cases, 17 had more than 20 metaphases: the 4 remaining cases did not contain any mitoses in any of the preparations. Four cases {cases 3, 4, 6 and 9} showed abnormal clones with trisomy 12 {Fig. 1). All these abnormal cases were examined shortly after diagnosis {1.2 months or less}. Three of these patients (cases 3, 4, and 6) were started on therapy 0.2, 1.0. and 9.2 months after diagnosis, respectively. On the other hand. the patients examined more than 5 months after diagnosis and who had not received any treatment (lid not show any abnormal clones: furthermore, most of them did not require any treatment for a long time after the cytogenetic examinations, as shown in Table 2. DISCUSSION Differences ill culture conditions and the PBA used by investigators [1, 4] may have contributed to the discrepancy about the incidence of B-CLL cases with abnormal chronlosomes. We have shown that 3-day cultures are optimal for routine cytogenetic studies in CLL. based on the data of the cell cv c|e of stimulated cells in BCLL using a sister chromatid exchange technique [7]: the most successful PBA are PWM, EBV, and LPS [8]. In the present study, the cells of 21 untreated B-CLL cases
Case
17, 18. 19. 20, 21,
F,B. R.McN. C.W. E.F. W.B.
16. J.G.
14. E.M. 15. C,W,
13. J.G.
E.O. W.A. J. McF. J.C. N.D.
7. J.G.
8. 9, 10. 11. 12.
58F 60F 68M 54M 74M 63M 73M 71M 76M 49M 74M 62F 61F 56M 63M 58F 70M 50M 56M 88F 83F
Age and sex
1. A.C, 2. F.M. 3. K.R. 4.].M. 5. J.B. 6. R.McC.
Hematological
Table 1
13.7 14.6 14.8 8.1 12.7 11.9 13.5 17.5 13.8 15.4 13.1 15.6 12.4 16.3 15.5 14.3 13.9 15.9 16.5 12,3 10.9
Hb (g/d|} 252 264 256 108 177 167 119 115 338 202 138 190 194 146 209 199 141 192 123 312 112
examination
32.0 176.0 26.5 16.6 64.7 10.3 21.3 20.0 55.6 16,0 45.5 16.4 49.3 33.9 25.4 42.0 21.9 12.5 9.8 33.1 6.1
W B C ( x 10a/ram :~)
data at the time of cytogenetic
P l a t e l e t s { x 10:~/mm a)
and immunological
89 77 64 83 92 66 83 73 80 65 92 78 6O 68 51 93 75 65 80 81 94
% of W B C 21 18 4 24 3 11 3 13 7 7 5 8 5 24 20 13 12 19 26 9 26
E
EA 39 22 28 12 15 5 15 3 50 28 5 56 26 14 31 34 not done 5 51 84 48
Lymphocytes
26 18 56 57 31
72
61 31 60 27 7 0 7 64 6 12 31 12 65 12 47
EAC
Kg Kg K~ KS~ K~
X~
K~ Xg X~g Xg XS~ Kg ~ ~ Kg K~g K~ K~ ~ ~ x~
Smlg
0.0 0.0 0.0 0.0 0.1 0.5 0.9 1.0 1.2 5.0 7.0 7.4 19.4 20.3 33.6 56.8 72.2 127.0 133.6 179.1 179.8
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
blood a nd m a r r o w blood blood blood blood a nd m a r r o w blood blood blood bl oo d blood marrow blood blood blood bl oo d blood blo od blood blood blood blood
Source
"Plus sign {+ ] indicates patient alive and still untreated.
T i m e from d i a g n o s i s to s a m p l i n g (months) 0 60 60 6O 0 60 60 60 60 23 20 25 23 0 34 20 39 20 0 21 25
to therapy
O (0) 0 {0) 33 (55) 19 (32) 0 (0) 4 (7) 0 (0) 0 (0) 29 (48) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0} 0 (0) 0 (0) 0 (0) 0 (0)
Abnormal metaphases (%)
and from sampling
Total n u m b e r of metaphases analyzed
C y t o g e n e t i c f i n d i n g s a n d t i m e f r o m d i a g n o s i s to s a m p l i n g
Case
T a b l e 2.
46,XY 47,XY. 47,XY, -47,XY. 46,XY 46,XY 47,XY, 46,XY 46,XY 46,XX 46,XX -46,XY 46,XX 46,XY 46,XY -46,XX 46,XX + 12
+ 12
+ 12 + 12
Karyotypes
11.9+ 5 .0 + 1.0 0.2 1.6 9,2 15.82.9 10.7 (dead) 18.6 + 16.6 + 1 5 .0 + 0.5 1 7 .9 + 1 8 .8 + 16.3 + 18.9± 8.4+ 16.9 16.2 + 18.9+
T i m e from s a m p l i n g to t h e r a p y a (months)
Figure 1
Karyotype of a lymphocyte stimulated by Epstein-Barr virus in case 3, showing an extra chromosome #12 (arrow).
50
N. Sadamori et al.
were examined under the above-mentioned conditions. Nevertheless, only four out of 21 (19%) cases showed an abnormal clone with trisomy 12. However, out of 9 cases where the patients were examined shortly after diagnosis, four {44.4%) showed abnormal clones. These four patients with abnormal ch r o m o so m es were started on therapy shortly after diagnosis or died shortly after cytogenetic examination and/or diagnosis. On the other hand, none of 12 cases (cases 10-21) where the patients were examined more than 5 months after diagnosis, w h i c h may indicate that they did not require treatment i mm ed i at el y after diagnosis, showed abnormal clones. Even though some exceptional cases, such as cases 8 and 13. where patients with normal karyotypes were started on therapy shortly after the cytogenetic examination, a correlation between the presence of an abnormal clone and the time from diagnosis to therapy seemed to exist. These facts suggest that the incidence of abnormal clones in untreated B-CLL cases may depend on their clinical status. On the other hand, 5 out of 41 treated B-CLL cases showed marker chromosomes, as reported by us [5]. These findings suggest the possibility that marker chromosomes in treated B-CLL cases [5] might be induced by c h e m o t h e r a p y and irradiation, or that the cells with marker chromosomes existed in low percentages before therapy but, possibly being resistant to the antileukemic drugs, were capable of proliferation after treatment. To resolve this question, the c h r o m o s o m e changes in the same patients before and after treatment should be compared. We found no correlation between the type of chromosome abnormalities and SmIg or between the value of EA- and EAC-rosettes and the percentage of cells with abnormal chromosomes in untreated B-CLL cases. A comparison of the relationship between the c h r o m o s o m e changes in B-CLL and the clinical and therapeutic aspects will be published and compared with those presented by other authors {11]. The authors wish to express their thanks to Mr. R. Holdsworth and Mrs. I. Kuttner for tedlnical assistance.
REFERENCES 1. Gahrton G, Robert KH, Friberg K, Zeck L. Bird AG (1980): Nonrandom chromosomal aberrations in chronic lymphocytic leukemia revealed by polyclonal B-cell-mitogen stimulation. Blood 56, 640-647. 2. Morita M, Minowada J, Sandberg AA {1981): Chromosomes and causation of hmnan can cer and leukemia. XIV. Chromosome patterns in stimulated lymphocytes of chronic: l~,mphocytic leukemia. Cancer Genet Cytogenet 3, 293-306. 3. Sadamori N, Han T, Minowada J, Matsui S, Sandberg AA (1982): Clonal chromosome changes in stimulated lymphocytes of untreated B-cell leukemias. Cancer Res (submitted). 4. Schroder J, Vuopio P, Autio K (1981): Chromosome changes in human chronic lymphocytic leukemia. Cancer Genet Cytogenet 4, 11-21. 5. Sadamori N, Han T. Minowada J, Sandberg AA (1982): Chromosome findings in treated patients with B-cell chronic lymphocytic leukemia. Cancer Genet Cytogenet (submittedl. 6. Minowada J, Janossy G, Greaves MF, Tsubota T, Srivastava BIS, Morikawa S, Tatsumi E (1978): The expression of antigen associated with acute lymphoblastic leukemia in human leukemia-lymphoma cell line. J Natl Cancer Inst 60, 1269-1277. 7. Sadainori N, Block AW, Han T, Sandberg AA (1982): Cell cycle analysis of stimulated lymphocytes of B-cell chronic lymphocytic leukemia. Cancer Genet Cytogenet (submittedl. 8. Sadamori N, Matsui S, Han T, Sandberg AA (1982); Comparative results with various polyclonal B-cell activators in anenploid chronic lymphocytic leukemia. Cancer (;enet Cytogenet (submitted).
U n t r e a t e d P a t i e n t s w i t h B-CLL
51
9. Sandberg AA (1980): The Chromosomes in Human Cancer and Leukemia. Elsevier/NorthHolland, New York. 10. First International Workshop on Chromosomes in Leukaemia (1978): Chromosomes in acute non-lymphocytic leukaemia. Br J Haematol 39, 311-316. 11. Rob6rt KH, Gahrton G, Friberg K, Zech L, Nilsson B (1982): Extra chromosome 12 and prognosis in chronic lymphocytic leukaemia. Scand J Haematol 28, 163-168.
ABSTRACT: The chromosome constitutions of stimulated lymphocytes in 21 untreated cases with Bcell chronic lymphocytic leukemia (B-CLL) were examined. So-called polyclonal B-cell activators, i.e., pokeweed mitogen, Epstein-Barr virus, and lipopolysaccharide W from E. coil 055:B5, were used. Four out of 21 cases showed abnormal clones with trisomy 12 and were started an therapy shortly after the diagnosis and cytogenetic examination. On the other hand, most cases without abnormal clones had not received treatment for relatively long periods before and after cytogenetic examination. These findings may indicate that cytogenetic results can be utilized as a parameter for treatment and prognosis in B-CLL.
INTRODUCTION Some investigators [1-3] have reported that the most c o m m o n c h r o m o s o m e abnormality of l y m p h o c y t e s stimulated by polyclonal B-cell activators (PBA) in B-cell chronic l y m p h o c y t i c leukemia (B-CLL) was trisomy 12. However, the i n ci d en ce of CLL cases with abnormal c h r o m o s o m e s differs from laboratory to laboratory. Schr6der et al. [4] found an abnormal clone in 1 out of 31 (3.2%) untreated B-CLL cases and in 4 out of 35 (11.4%) treated cases, including those without any metaphases. Gahrton et al. [1] reported that 8 out of 11 (72.7%) untreated and treated cases of B-CLL showed abnormal clones. Morita et al. [2] fou'~d abnormal clones in 9 out of 27 (33.3%) cases, including those w i t h o u t any metaphases. The differences in the i n ci d e n c e of abnormal clones among the B-CLL cases r\eported to date may be related to such factors as stage of disease, interval betwee~ the diagnosis and cytogenetic examination and treatment background. We have already reported that 4 out of 9 (44.4%) untreated and n e w l y diagnosed B-CLL cases [3] and 15 out of 40 (37.5%) treated B-CLL cases showed abnormal clones [5]. In the present study, we attempted to clarify the i n ci d en ce of abnormal clones among untreated cases and the relationship between chromosomal abnormalities and some clinical parameters.
From the Roswell Park Memorial Institute, Buffalo. Address requests for reprints to Avery A. Sandberg, M.D., Roswell Park Memorial Institute, 666 Elm Street, Buffalo, N Y 14263. Received December 1, 1962; accepted March 3, 1983.
45 '~ 1984 by Elsevier Science Publishing Co., Inc. 52 Vanderbilt Ave., New York, NY 10017
Cancer Genetics and Cytogenetics 11, 45 51 (1984) 0165-4608/84/$03.00
46
N. Sadamori et al.
MATERIALS AND METHODS Lymphocytes were obtained from 21 untreated cases with clinically and imnlul~ologically typical B-CLL (14 males and 7 females], whose ages ranged from 49 to 88 years, the mean age being 63 years. The diagnoses were based on accepted clinical and laboratory criteria. Clinical, hematological, and immunological data on the cases are shown in Table 1. No patient in the present study had a history of a¢:cidental or therapentic irradiation or therapy with anticancer drugs. Nine cases leases 1-9) have been already reported hy us 131. The methodologies for E-rosette, EA-rosette, and EAC-rosette formation and surface membrane i m m u n o g l o b u l i n s (SmIg) of the separated lymphocytes have been reported by M i n o w a d a et al. 161. Lymphocytes were separated from 20 ml of heparinized peripheral blood or 2 ml of heparinized bone marrow material by centrifugation over a Ficoll-Hypaque gradient. Separated lymphocytes {1 × 10 '~ cells/roll were cultured for 3 days and harvested according to procedures described by us previously [7, 81. Usually 10-ml cultures were set up: three cultures with pokeweed mitogen (PWM), Epstein-Bari: virus (EBV), and lipopolysaccharide W from E. coli 055:B5 {LPS) were set up t()r each case. Karyotype analyses were performed by G- and Q-banding techniques I,~l. At least 20 optimally banded metaphases were analyzed in each case. An abnormal clone was defined as the presence of two cells with the same extra chromosome or structural rearrangement, or three cells with the same missing chromosome, according to the First International Workshop on Chromosomes in Leukemia [10 I. Treatment {chlorambucil and/or prednisone) was given to all B-CI,i, patients with stage [II-IV disease, as well as to stage [-II patients when their splenomegaly and/or l y m p h a d e n o p a t h y significantly increased. No treatment was usually necessary in patients with stage 0 disease. Case 3 with stage O, associated with monoclonal IgM gammopathy (3,500 mg/ml), was treated with chlorambucil because c)f the d e v e l o p m e n t of peculiar neurological symptoms suggestive of a hyperviscosity syndrome.
RESULTS As shown in Table 2, out of the 21 cases, 17 had more than 20 metaphases: the 4 remaining cases did not contain any mitoses in any of the preparations. Four cases {cases 3, 4, 6 and 9} showed abnormal clones with trisomy 12 {Fig. 1). All these abnormal cases were examined shortly after diagnosis {1.2 months or less}. Three of these patients (cases 3, 4, and 6) were started on therapy 0.2, 1.0. and 9.2 months after diagnosis, respectively. On the other hand. the patients examined more than 5 months after diagnosis and who had not received any treatment (lid not show any abnormal clones: furthermore, most of them did not require any treatment for a long time after the cytogenetic examinations, as shown in Table 2. DISCUSSION Differences ill culture conditions and the PBA used by investigators [1, 4] may have contributed to the discrepancy about the incidence of B-CLL cases with abnormal chronlosomes. We have shown that 3-day cultures are optimal for routine cytogenetic studies in CLL. based on the data of the cell cv c|e of stimulated cells in BCLL using a sister chromatid exchange technique [7]: the most successful PBA are PWM, EBV, and LPS [8]. In the present study, the cells of 21 untreated B-CLL cases
Case
17, 18. 19. 20, 21,
F,B. R.McN. C.W. E.F. W.B.
16. J.G.
14. E.M. 15. C,W,
13. J.G.
E.O. W.A. J. McF. J.C. N.D.
7. J.G.
8. 9, 10. 11. 12.
58F 60F 68M 54M 74M 63M 73M 71M 76M 49M 74M 62F 61F 56M 63M 58F 70M 50M 56M 88F 83F
Age and sex
1. A.C, 2. F.M. 3. K.R. 4.].M. 5. J.B. 6. R.McC.
Hematological
Table 1
13.7 14.6 14.8 8.1 12.7 11.9 13.5 17.5 13.8 15.4 13.1 15.6 12.4 16.3 15.5 14.3 13.9 15.9 16.5 12,3 10.9
Hb (g/d|} 252 264 256 108 177 167 119 115 338 202 138 190 194 146 209 199 141 192 123 312 112
examination
32.0 176.0 26.5 16.6 64.7 10.3 21.3 20.0 55.6 16,0 45.5 16.4 49.3 33.9 25.4 42.0 21.9 12.5 9.8 33.1 6.1
W B C ( x 10a/ram :~)
data at the time of cytogenetic
P l a t e l e t s { x 10:~/mm a)
and immunological
89 77 64 83 92 66 83 73 80 65 92 78 6O 68 51 93 75 65 80 81 94
% of W B C 21 18 4 24 3 11 3 13 7 7 5 8 5 24 20 13 12 19 26 9 26
E
EA 39 22 28 12 15 5 15 3 50 28 5 56 26 14 31 34 not done 5 51 84 48
Lymphocytes
26 18 56 57 31
72
61 31 60 27 7 0 7 64 6 12 31 12 65 12 47
EAC
Kg Kg K~ KS~ K~
X~
K~ Xg X~g Xg XS~ Kg ~ ~ Kg K~g K~ K~ ~ ~ x~
Smlg
0.0 0.0 0.0 0.0 0.1 0.5 0.9 1.0 1.2 5.0 7.0 7.4 19.4 20.3 33.6 56.8 72.2 127.0 133.6 179.1 179.8
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
blood a nd m a r r o w blood blood blood blood a nd m a r r o w blood blood blood bl oo d blood marrow blood blood blood bl oo d blood blo od blood blood blood blood
Source
"Plus sign {+ ] indicates patient alive and still untreated.
T i m e from d i a g n o s i s to s a m p l i n g (months) 0 60 60 6O 0 60 60 60 60 23 20 25 23 0 34 20 39 20 0 21 25
to therapy
O (0) 0 {0) 33 (55) 19 (32) 0 (0) 4 (7) 0 (0) 0 (0) 29 (48) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0} 0 (0) 0 (0) 0 (0) 0 (0)
Abnormal metaphases (%)
and from sampling
Total n u m b e r of metaphases analyzed
C y t o g e n e t i c f i n d i n g s a n d t i m e f r o m d i a g n o s i s to s a m p l i n g
Case
T a b l e 2.
46,XY 47,XY. 47,XY, -47,XY. 46,XY 46,XY 47,XY, 46,XY 46,XY 46,XX 46,XX -46,XY 46,XX 46,XY 46,XY -46,XX 46,XX + 12
+ 12
+ 12 + 12
Karyotypes
11.9+ 5 .0 + 1.0 0.2 1.6 9,2 15.82.9 10.7 (dead) 18.6 + 16.6 + 1 5 .0 + 0.5 1 7 .9 + 1 8 .8 + 16.3 + 18.9± 8.4+ 16.9 16.2 + 18.9+
T i m e from s a m p l i n g to t h e r a p y a (months)
Figure 1
Karyotype of a lymphocyte stimulated by Epstein-Barr virus in case 3, showing an extra chromosome #12 (arrow).
50
N. Sadamori et al.
were examined under the above-mentioned conditions. Nevertheless, only four out of 21 (19%) cases showed an abnormal clone with trisomy 12. However, out of 9 cases where the patients were examined shortly after diagnosis, four {44.4%) showed abnormal clones. These four patients with abnormal ch r o m o so m es were started on therapy shortly after diagnosis or died shortly after cytogenetic examination and/or diagnosis. On the other hand, none of 12 cases (cases 10-21) where the patients were examined more than 5 months after diagnosis, w h i c h may indicate that they did not require treatment i mm ed i at el y after diagnosis, showed abnormal clones. Even though some exceptional cases, such as cases 8 and 13. where patients with normal karyotypes were started on therapy shortly after the cytogenetic examination, a correlation between the presence of an abnormal clone and the time from diagnosis to therapy seemed to exist. These facts suggest that the incidence of abnormal clones in untreated B-CLL cases may depend on their clinical status. On the other hand, 5 out of 41 treated B-CLL cases showed marker chromosomes, as reported by us [5]. These findings suggest the possibility that marker chromosomes in treated B-CLL cases [5] might be induced by c h e m o t h e r a p y and irradiation, or that the cells with marker chromosomes existed in low percentages before therapy but, possibly being resistant to the antileukemic drugs, were capable of proliferation after treatment. To resolve this question, the c h r o m o s o m e changes in the same patients before and after treatment should be compared. We found no correlation between the type of chromosome abnormalities and SmIg or between the value of EA- and EAC-rosettes and the percentage of cells with abnormal chromosomes in untreated B-CLL cases. A comparison of the relationship between the c h r o m o s o m e changes in B-CLL and the clinical and therapeutic aspects will be published and compared with those presented by other authors {11]. The authors wish to express their thanks to Mr. R. Holdsworth and Mrs. I. Kuttner for tedlnical assistance.
REFERENCES 1. Gahrton G, Robert KH, Friberg K, Zeck L. Bird AG (1980): Nonrandom chromosomal aberrations in chronic lymphocytic leukemia revealed by polyclonal B-cell-mitogen stimulation. Blood 56, 640-647. 2. Morita M, Minowada J, Sandberg AA {1981): Chromosomes and causation of hmnan can cer and leukemia. XIV. Chromosome patterns in stimulated lymphocytes of chronic: l~,mphocytic leukemia. Cancer Genet Cytogenet 3, 293-306. 3. Sadamori N, Han T, Minowada J, Matsui S, Sandberg AA (1982): Clonal chromosome changes in stimulated lymphocytes of untreated B-cell leukemias. Cancer Res (submitted). 4. Schroder J, Vuopio P, Autio K (1981): Chromosome changes in human chronic lymphocytic leukemia. Cancer Genet Cytogenet 4, 11-21. 5. Sadamori N, Han T. Minowada J, Sandberg AA (1982): Chromosome findings in treated patients with B-cell chronic lymphocytic leukemia. Cancer Genet Cytogenet (submittedl. 6. Minowada J, Janossy G, Greaves MF, Tsubota T, Srivastava BIS, Morikawa S, Tatsumi E (1978): The expression of antigen associated with acute lymphoblastic leukemia in human leukemia-lymphoma cell line. J Natl Cancer Inst 60, 1269-1277. 7. Sadainori N, Block AW, Han T, Sandberg AA (1982): Cell cycle analysis of stimulated lymphocytes of B-cell chronic lymphocytic leukemia. Cancer Genet Cytogenet (submittedl. 8. Sadamori N, Matsui S, Han T, Sandberg AA (1982); Comparative results with various polyclonal B-cell activators in anenploid chronic lymphocytic leukemia. Cancer (;enet Cytogenet (submitted).
U n t r e a t e d P a t i e n t s w i t h B-CLL
51
9. Sandberg AA (1980): The Chromosomes in Human Cancer and Leukemia. Elsevier/NorthHolland, New York. 10. First International Workshop on Chromosomes in Leukaemia (1978): Chromosomes in acute non-lymphocytic leukaemia. Br J Haematol 39, 311-316. 11. Rob6rt KH, Gahrton G, Friberg K, Zech L, Nilsson B (1982): Extra chromosome 12 and prognosis in chronic lymphocytic leukaemia. Scand J Haematol 28, 163-168.