Chromosome 14q+ in Adult T-Cell Leukemia

Isao Miyoshi, Kanji Miyamoto, Mitsuo Sumida, Ryuji Nishihara, Mfinyuh Lai, Shizuo Yoshimoto, liro Sato, and Ikuro Kimura

ABSTRACT: Cytogenelic studies were performed on leukemic cells from two patients with adult T.cell leukemia. A 14q+ marker chromosome was Found in the peripheral blood leukocytes [ram pa- tient No. 1 and in a leukemic T-cell line (MT.I) derived From the peripheral blood o/patient No. 2. The 14q+ resulted from o t(12:14) in patient No. I and from a tfY:14) in patient No. 2 with o break point at 14q32 in each case. In addition, the leukemic cells from patient No. ! showed o t(1;7) and a 9q--, while the MT.I line had numerous structural abnormalities. Thus, it is clear that a 14q + tronslocation is not restricted to B-cell neoplasms but occurs in T.cell neoplasms as well.

INTRODUCTION

A 14q+ marker chromosome has been observed in a variety of lymphoid malignan- cies, such as Burkitt's and non-Burkitt's lymphoma [1-5], multiple myeloma (plasma-cell leukemia) [6,7], and acute lymphoblastic leukemia (ALL) of B-cell or Burkitt type [8-10]. These disorders are predominantly B-cell cancers, and the 14q+ chromosome has been infrequently associated with T-cell malignancies [11-15]. We report here two patients with adult T-cell leukemia (ATL), in whom this marker chromosome was found.

CASE RI~'ORT

Patient No. 1 (N.K.), a 3e-year-old man, was admitted in November 1978 with jaun- dice, generalized lymphadenopathy and hepatosplenomegely. The patient first noted lymph node swelling in April 1978 but sought no medical attention because of th~ lack of other symptoms. The WBC count was 74,000/mm 3 with 86% lymphocytes and the platelet count 59,000/m_m s. There was no anemia. The majority of the lympho- cytes were abnormal with indented or lobulated nuclei. The nucleoli were generally indistinct. A bone marrow aspirate contained 41% lymphocytes, more than half of which appeared abnormal. The Ficoll-Conray-separated fraction of peripheral blood leukocytes contained 82% E-rosette-forming T-cells (Fig. 1). A cervical lymph node biopsy revealed pleomorphic lymphoma with Reed-Sternber8 type giant cells (Fig. 2). Chest X-ray films showed no mediastinal mass. Treatment with L-asparaginase,

From the Depeurtment of Medicine and the Cancer Institute Division of Pathology, Okayama University Medical School, Okay~_m_R0 Japan.

Address requests for reprints to: Dr. Isao Miyoshi, Department of Medicine, Okayama Uni- versiiy Medical School, Okayama 700, Japan.

Received June 17, 1980; accepted November 10, 1980.

251 © Elsevier North Holland, Inc., 1981 52 Vanderbilt Ave., New York. NY 10017

Cancer Genetics and Cytogenetics 3, 251-259 (1981) 0165-4608181/032510952.50

252

Figure I Peripheral blood abnormal lymphocytes from patient No. 1 forming spontaneous E-rosettes. Note the nuclear indentation or lobulation. IVlay-Griinwald-Giemsa stain, x 1,000.

Figure 2 Lymph node b~opsy from patient No. 1 showing pleomorphic lymphoma with Reed-Sternberg type giant cells. Hematoxylin and eosin stain, ×250.

Chromosome 14q+ in Adult T-Cell Leukemia 253

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Fisqutre 3 Peripheral blood abnormal lymphocytcs from patient No. 2 showing indented or lobulated nuclei. May-Griinwald-Giemsa stain, Xl,000.

vincristine, and prednisolone induced a partial remission with clearing of the jaun- dice. In January 1979, another course of the same chemotherapy regimen was started. However, interstitial pneumonitis and generalized herpes zoster infection developed and the patient died on February 17, 1979. Autopsy revealed leukemic infiltration in the 13unph nodes, liver, spleen, lungs, and kidneys. The bone marrow was hypo- plastic.

Patient No. 2 (M.T.), a 69-year-old man, was admitted in May 1978 with malaise

Figure 4 Cells from the MT-1 line showinR nuclear irregularities. May-Griinwald-Giemsa stain, Xl,000.

254 I. Miyoshi et al.

and generalized lymphadenopathy. There was no hepatosplenomegaly. The skin was normal. The WBC count was 13,000/mm :~ with 21% lymphocytes and 41% abnormal lymphocytes (Fig. 3). The nuclei of the abnormal lymphocytes were, as in patient No. 1, irregularly shaped with indentation or lobulation. The chromatin was mostly con- densed with indistinct nucleoli. The hemoglobin was 12.9 g/dl and the platelet count 132,000/mm:". Surface marker studies showed 79% of the peripheral mononuclear cells to be E-rosette-forming T-cells. An inguinal lymph node biopsy specimen was interpreted as malignant lymphoma. Chest X-ray films were normal. The patient was treated with vincristine, cyclophosphamide, 6-mercaptopurine, and prednisolone with a partial regression of the lymph nodes. Despite continued chemotherapy, the lymph nodes began to enlarge within two months and the peripheral leukocytes gradually increased to 183,000/mm :~ with 88% abnormal lymphocytes. A bone mar- row sample aspirated 2 days before death contained 63.5% abnormal lymphocytes. The patient died with hypercalcemia 4 months after onset. Postmortem examination revealed leukemic infiltration in various organs including the lymph nodes, liver, spleen, lungs, kidneys, bone marrow, and gastrointestinal tract. A continuous culture l i ne of leukemic 'r-cells (MT- 11 was established from a peripheral blood sample taken 1 week before death. The MT-1 cells formed E-rosettes and exhibited nuclear irregu- larities {Fig. 4). The establishment and characteristics of this cell line have been re- ported [16,171.

CYTOGENETIC STUDIES

Chromosome analysis was performed on peripheral blood leukocytes obtained from patient No. 1 prior to chemotherapy. The leukocytes were separated on a Ficoll-Con- ray gradient and cultured for 24 hr in RPMI 1840 medium supplemented with 20% fetal calf serum. No phytohemagglutinin was added. Chromosomes of the MT-1 line derived from patient No. 2 were analyzed 140 days after initiation of the culture [3 weeks after establishment of the cell line). The cells from either source were incubat- ,d for 2 hr in the presence of Colcemid (0.5 ttg/ml), treated with 0.075 M KC! for 13 min at 37C. and fixed in a methyl alcohol :acetic acid {3:1) solution. Air-dried chro- mosomes were stained with c:onventional Giemsa and banded with quinacrine mus- tard {181 and trypsin-(;iemsa [191. Suitable metaphases were photographed and karyotyped.

RESULTS

leorty-eight of fifty cells from peripheral blood of patient No. 1 had 46 chromosomes. Of 15 (:ells analyzed with banding techniques, 3 were normal and 12 pseudodip- loid. All the abnormal (:ells had an identical karyotype: 46,XY,t(1;7){p36;q22), del(9}(q12),li12;14)(q13;q32) [Fig. 5).

The MT-1 line derived fro,a patient No. 2 had 45 chromosomes in 48 of 50 meta- phases. Banding analysis of 25 cells revealed numerous complex rearrangements and aberrations. A representative karyotype was: 45,-X, dup Yq,-13,-17, del(2)[q33],t{3; 131(q21;q34),int tlel[~){ql6 or q22),8q+,9p+,9q+,10q+,14p+.t{Y;14)[q12;q32).16q+. 18q+,+mar 1,÷mar 2 iFig, 61, There were minor variations among these cells but the. majority of th~ rearrangements, including the 14q+, were present in common in all the (:ells analyzed, The t(Y;14) arose from an inverted translocation of Yq to the e, nd of 14q, The duplicated segment on Yq appeared to be p l e r ~ q l 2 ::ql2--,ql 1. One of the two marker chromosomes probably contained the X chromosome. These results are summarized in Table 1.

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Chromosome 14q÷ in Adult T-Cell Leukemia 257

Table 1 Cytogenetic studies on two patients with ATL

Case number Age/sex Material

Chromosome number ' 44 45 46 4n Karyotype

1 36/M Peripheral blood

2 69fl~4 Cell l inp 2 48[25)

48{15) 2 46,XY.t{1:7}(p36:q22}.del{g}(q 12). t|12;14){q13;q32)

45,-X,dupYq,- 13,-17,del(2}{q33 }. t(3;13}{q21;q34),int del{6}{ql6 ur q22),Sq+.9p+.9q+. 10q+./4p4. t{Y: 14){qi 2:q32), 16q+.t 8q-~, +mar I. +mar 2

°Number in parentheses = number of banded cells examined.

DISCUSSION

In the present investigation, we have demonstrated a 14q+ in leukemic cells from two patients with ATL. The peripheral leukocytes from one patient carried a t(12:14} and other chromosome abnormalities. A leukemic T-cell line {MT-1) derived from the other patient had numerous structural abnormalities including a t(Y:14}. Because of the complex nature of the rearrangements, some of the abnormalities could not be fully identified. In each case, the break point in chromosome #14 was at 14q32. Since the chromosomes of leukemic cells from the donor of the MT-I line were not directly examined, it is difficult to be certain that the karyotype observed in the de- rived cell line truly represents what might have been present in vivo. The cytogenetic analysis was performed 140 days after initiation of the culture and it is possible that some of the abnormalities occurred under the in vitro conditions.

In Burkitt's lymphoma and Burkitt-type ALL, a 8q-;14q+ translocation has been consistently observed irrespective of African or non-African origin [ I - 4 , 10]. This marker chromosome is not related to the presence or absence of Epstein-Barr virus. Recently, the reciprocal nature of the t(8;14) has been determined by prophase-pro- metaphase chromosome analysis [20, 21]. In other lymphoid malignancies, mostly of the B-cell type, however, the donor chromosome involved in the genesis of a 14q4 is not consistent, but the recipient is chromosome #14. usually at 14q32 and rarely at 14q13 [22]. In contrast, 14q+ has been found infrequently in T-cell malignancies. It has been reported in two cases ofT-cell chronic lymphocytic leukemi~ (CLL) [I I ], one case of mycosis fungoides [12], and three cases of T-cell leukemia superimposed on ataxia telangiectasia [13-15]. In all these cases also, the morphology of the 14q+ was different from the typical t{8;14) of Burkitt-type lymphoma. The three ataxia telan- giectasia patients with T-cell leukemia exhibited a characteristic 14;14 translocation and in two of them the leukemic cells evolved from the pre-existing ataxia telangiec- tasia clone that had the same translocation [13, 15].

The morphology of the 14q + herein described in our two patients appears to be unusual with respect to the donor chromosomes. In various lymphoid neoplasms (other than Burkitt's lymphoma) with a 14q+, chromosomes #1, #8, #11, #14, and #18 are predisposed to act as donor chromosomes. Recently, an American Burkitt's lymphoma with two distinct karyotypes, a t(8;14) and a t(12;14), has been reported [4]. This is the only reported case in which a t(12;14), apparently similar to that ob- served in one of our patients, was found. A 14q+ involving a translocation from Yq. as identified in the MT-I line, has not been described. In a preliminary report, Ueshi- ma et al. [23] described a 14q+ in three patients with ATL. In their study, however, the origin of the 14q+ was not determined. Their finding as well as ours suggest that ATL may be a disease often associated with a 14q+ lransiocation.

ATL is a recently reported T-cell malignancy in Japan, and the clustering of the

2 5 8 I. Miyoshi et al.

patient's birthplaces in Kyushu was considered noteworthy [24]. The disease is char- acterized by onset in adulthood, subacute or chronic course, leukemic T-cells with irregularly shaped nuclei and lack of mediastinal masses. Sporadic cases of ATL have since been reported among people who were bern in other places and indeed both of our patients were born and lived in Shikoku, an island adjacent to Kyushu. It is of interest to note that the incidence of ATL is higher than that of B-cell CLL in Ja- pan, whereas the situation appears to be reversed in Europe and the United States. The clinical and cytopathologic features of ATL resemble in part those reported for T-cell CLL [25], small lymphocyte T-cell leukemia [26], malignant lymphoma of pe- ripheral T-lymphocyte origin [27], and T-cell malignant lymphoma [28]. We have presented immunologic evidence suggesting the peripheral T-cell origin of ATL cells [29]. The identity and classification of these T-cell neoplasms remain to be deter- mined.

This work was supported in part by a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan. The authors thank Dr. Janet D. Rowley for help with the karyotypic analysis and for reviewing this manuscript. They also thank Dr. Hirokuni Taguchi for providing a pe- ripheral blood sample of patient No. 2 and Drs. Shoichi Yamashita and Katsumi lwata for au- topsy findings.

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