Adult T-cell leukemia cells expressing OKT 17 antigen in the activated state

The surface phenotype and the functional activities of leukemic cells from seven patients with adult T-cell leukemia (ATL) were studied using monoclonal antibodies OKT 3, 4 and 8, anti-Tac, and OKT 17. The latter defines the heterogeneity of the activated T4+ T cell subset. In all cases, ATL cells with the typical OKT3-T4+ T8- phenotype expressed OKT 17 antigen. In addition, in five out of the seven cases the fresh ATL cells possessed Tac antigen which is expressed on activated T cells in varying degree. After cultivation with PWM, most populations of ATL cells acquired Tac even in the cases expressing little antigen in uncultured preparations. However the PWM activated ATL cells did not lose OKT 17 antigen. Functional assays showed the suppressor activity of ATL cells on normal B cell differentiation in three out of six cases examined. These results suggest that ATL cells most probably arise from a particular subset characterized by OKT 17 antigen within the activated OKT4+ T cell subset.     

Virus associated adult T-cell leukemia (ATL) in Japan: clinical, histological and immunological studies

Virus associated adult T-cell leukemia/lymphoma (ATLL), which includes both adult T-cell leukemia (ATL) and its non-leukemic counterpart (NLATL) was studied clinically, histologically, and immunologically. The disease usually occurred in the sixth decade in both sexes equally. The patients had a rapid clinical course with frequent leukemic changes, lymphadenopathy, hepatomegaly, and occasional skin rash. Bone marrow involvement with mild infiltration and hypercalcemia were more frequent in ATL than in NLATL. Histologically the disease was categorized as malignant lymphoma, diffuse pleomorphic type with cerebriform nuclear giant cells. The lymphoma was characterized by diffuse proliferation of tumor cells with irregular nuclear configurations, varying in size and shape, and the presence of giant cells with highly convoluted cerebriform nuclei. The giant cells seemed to be a diagnostic marker. Immunologically, the tumor cells usually possessed the surface antigens recognized by OKT 3, 4, Leu 8 and anti-Tac antibodies, indicating that they were lymphomas of helper/inducer peripheral T-cells with the receptor for interleukin 2, but they demonstrated no helper/inducer functions. The patients often died of opportunistic infections due to T-cell dysfunction caused by the disease itself and strong chemotherapy.     

Unusual Morphological Features of Adult T-cell Leukemia Cells with Aberrant Immunophenotype

We describe 4 cases of adult T-cell leukemia (ATL) with unusual morphology and aberrant immunophenotype. All patients were Japanese and born in the Nagasaki district, an area endemic for HTLV-I. Peripheral blood and/or bone marrow films revealed bizarre giant cells with and without large nucleoli; the cells were 5 to 6 times the diameter of erythrocytes, resembling Hodgkin's cells. Some peripheral blood cells were morphologically similar to prototypic ATL cells, while many other cells in the bone marrow showed unusual morphology. Furthermore, leukemic cells had aberrant immunophenotypes such as the CD8-positive type in patients 1 and 2, the CD4^- ˙ CD8^- double-negative type in patient 3, and the CD5 antigen defect in patient 4. All patients had marked elevations of the serum calcium and LDH and organomegaly, while all had a short survival. Anti-HTLV-I antibodies and provirus DNA monoclonality were demonstrated in all patients.

The results suggested that the unusual morphology and aberrant ATL cell immunophenotype may be indicative of a high grade malignant behaviour of ATL.     

Immunoregulatory function and expression of OKT17 antigen of adult T-cell leukemia cells

Research was carried out on neoplastic T-cells taken from 10 cases of Japanese adult T-cell leukemia (ATL) in regard to their immunoregulatory activity on pokeweed mitogen (PWM)-driven immunoglobulin (Ig) production and their differentiation antigens. ATL cells were reactive with OKT3, OKT4 and OKT11 in most cases; however, the ATL cells from one case did not react with OKT11 nor formed E-rosettes. ATL cells of three cases were reactive with OKT8 as well as OKT4. In six cases, the ATL cells were reactive with OKT17 and coincidentally displayed suppressor activity on Ig production. The cells of the remaining four cases did not have such reactivity with OKT17 or suppressor activity. Moreover, the OKT17(+) ATL cells from one case sorted by FACS 440 demonstrated suppressor activity, while the OKT17(-) ATL cells did not. These results indicated that ATL cells with suppressor activity had OKT3(+), OKT4(+), OKT17(+) phenotype.     

PRODUCTION OF PARATHYROID HORMONE-RELATED PROTEIN IN ADULT T-CELL LEUKEMIA CELLS

Human parathyroid hormone-related protein (PTHrP) mRNA was detected in peripheral leukemic cells obtained from adult T-cell leukemia (ATL) patients as well as in cultured human T-cell leukemia virus type I (HTLV-D-infected T-cell lines. In contrast, PTHrP mRNA was not detected in other types of leukemic cells. Using radioimmunoassay, immunoreactive PTHrP was also detected in the spent media of HTLV-I-infected T-cell lines. These results suggest that PTHrP plays an important role in developing the hyperealcemia frequently observed in ATL patients.     

IgG-κ-Type Plasmacytoma Secreting Salivary-Type Amylase in Adult T-Cell Leukemia

A IgG-κ-type plasmacytoma secreting salivary-type amylase ectopically is reported in a patient with smouldering adult T-cell leukemia(ATL). The patient had plasmacytomas in the distal region of the right femur, the proximal region of left tibia, and the left paranasal sinus. Both his serum and urine contained high levels of amylase. The presence of IgG-κ and S-type amylase in the plasmacytoma cells was confirmed immunocytochemically. In addition, he was also positive for the antibody against the human T-cell leukemia virus type I (HTLV-I), and had abnormal lymphocytes with convoluted nuclei(ATL cells) in the peripheral blood. The monoclonal integration of HTLV-I proviral DNA was demonstrated in the leukemic cells of the peripheral blood, but not in the plasmacytoma cells. Our case suggested that not only can HTLV-I infection play a role in the development of ATL, but may also induce a B-cell malignancy in an indirect manner, and even an ectopic amylase producing plasmacytoma.     

A case of CD4+/CD8− adult T-cell leukemia with good response to interferon-beta terminating as a CD4+/CD8+ adult T-cell lymphoma

The leukemic cells of adult T-cell leukemia (ATL) usually express the helper/inducer associated antigen reactive with anti-CD4 antibodies but not with anti-CD8. We present a 63-yr-old woman with ATL characterized by circulating leukemic cells with CD4+/CD8− phenotype, hepatosplenomegaly with no lymphadenopathy, and the presence of proviral DNA of human T-cell leukemia virus I in the leukemic cells. She was successfully treated with interferon beta and the remission lasted for 12 months. She then relapsed in the lymph nodes with minimal peripheral blood involvement. The neoplastic cells of the lymph node now co-expressed CD4 and CD8 antigens indicating that the change in clinical manifestation was accompanied by a phenotypic change of the leukemic cells.     

Childhood acute lymphoblastic leukemia with natural killer activity. Clinical and cellular features of three cases

Leukemic cells from 3 of 28 children with acute lymphoblastic leukemia (ALL) had natural killer (NK) activity against K562 and Molt-3, but not against Raji as determined by the 4-hour 51Cr release assay at a 40:1 effector:target ratio; the highest percent lysis against K562 in each patient ranged from 55.9% to 147.4% of the normal lymphocyte value and that against Molt-3 from 28.0% to 127.9%. Their leukemic cells were nonphagocytic, nonadherent, and negative for nonspecific esterase. Leukemic cells from two of them displayed similar morphologic and immunologic features. The cells had a round nucleus, and were E-receptors (E)+, Leu-5+, IgG-Fc+, Ia+, OKM1+ or OKM1-, T-cell antigens-, B markers-, and monocyte antigen-, indicating their NK cell origin. The other patient's cells were characterized by the irregularly shaped nucleus, and were E+, Leu-5+, IgG-Fc+, T+, OKT6+, Leu-2a+, and OKM1+. The presence of E which are identical to E of T-cells and a high density of T-cell antigens including OKT6 on the cells suggested their T-cell nature. Of the three patients, two without thymic enlargement had leukemic cells of NK cell origin, and the other one with the symptom leukemic cells of the T-cell lineage, the clinical feature probably reflecting the cell lineage of their leukemic cells.     

Cytogenetic studies on the adult T-cell leukemia in Japan

Cytogenetic studies were performed on 16 patients with ATL seen in Northern Kyushu island; nine were patients with acute type leukemia, one with crisis type and five with lymphoma type. The serum antibody for HTLV-1 (ATLA) was positive in all patients and the phenotype of ATL cells were ERFC+, OKT3+, OKT4+, OKT6-, OKT8-, OKT10+, OKla1+/- and Tac+. Abnormal findings of chromosomes were observed in 15 patients. Thirteen patients were in near diploid range. One patient was in triploid range and one patient was in tetraploid range. The polyploid karyotypes were found only in lymphoma type patients. Trisomy 3 and trisomy 7 were observed each in three patients with acute type of ATL. The most frequent abnormal rearrangement was observed in the long arm of chromosome 6 and the break occurred at band 6q15 and 6q21 each in four patients in this series.     

The Differences between Lymphoma and Leukemia Type of Adult T-cell Leukemia

The difference between lymphoma type and leukemia type of adult T-cell leukemia (ATL) were analysed with 102 Japanese patients all positive for human T-cell leukemia virus type I (HTLV-I) antibody. They were classified into three groups on findings at first medical examination: lymphoma type cases, leukemia type cases, and mixed type (leukemia type plus lymphadeno-pathy) cases. Lymphoma type patients had several or more enlarged lymph nodes the largest of which was greater than 1 cm in diameter and with practically no abnormal lymphocytes (ATL. cells), which are characteristic of ATL, in the peripheral blood. Leukemia type patients had 10% or more ATL cells in the peripheral blood and had no detectablle lymphadenopathy Lymphoma type patients often complained of detectable lymphadenopathy, while leukemia type patients complained frequently of general fatigue and skin eruption. Mixed type patients more frequently had signs and symptoms which were characteristic of both types: lymphadenopathy and 10% or more ATL cells in the peripheral blood. Mixed type: ATL had a poorer prognosis than either lymphoma type or leukemia type. The median survival time was 3 months for mixed type patients, 10.5 months for lymphoma type patients, and 13.5 months for leukemia type patients. Complications and causes of death have also been touched upon. Clinicians are thus advised to consider ATL patients separately according to their clinical manifestations.     

Serum Soluble Interleukin-2 Receptor Levels in Patients with Adult T-cell Leukemia and Human T-cell Leukemia/Lymphoma Virus Type-I Seropositive Healthy Carriers

Using an enzyme-linked immunosorbent assay (ELISA) technique, we measured the soluble interleukin 2 receptor (s-IL-2R) levels in the sera of patients with adult T-cell leukemia (ATL) in Japan. The s-IL-2R levels in the sera of the ATL patients were markedly higher (range 540-310, 400 U/ml, mean ±SD=62,800 ±81,000 U/ml, n = 42) than those in normal individuals (range 42-950 U/ml, mean ±SD=322 ±198 U/ml, n = 35, P<0.01). The patients with acute-type or lymnhoma-type ATL had high s-IL-2R levels (range 11,900-310,400 U/ml, mean ±SD= 110,340 ± 370 U/ml, n = 15; range 26,400-214,400 U/ml, mean ±SD=90,170 ±59,040 U/ml, n = 7, respectively). All of the patients with hypercalcemia (Ca>10 mg/dl) or elevated serum LDH levels (LDH > 500 IU/liter) also had s-IL-2R levels above 10,000 U/ml. The high s-IL-2R levels in the sera of ATL patients indicate abnormal IL-2 receptor production and its release from the leukemic cells in vivo . Thus, the serum s-IL-2R level may be a sensitive and useful marker to monitor the total amount of tumor cells in ATL, especially in the lymphoma type. We next examined the serum s-IL-2R levels in human T-cell leukemia/lymphoma virus type-I (HTLV-I) seropositive healthy carriers to investigate whether there might he abnormal IL-2 receptor expression in such individuals. However, there was no statistically significant difference between the S-IL-2R level of 71 HTLV-I seropositive healthy carriers (range 65-880 U/ml, mean±SD =394±212 U/ml) and that of 71 age- and sex-matched normal individuals (range 33-950 U/ml, mean ±SD=357 ±224 U/ml) who lived in Okinawa Prefecture.     

Clinicopathological Features of Adult T-Cell Leukemia with CD30 Antigen Expression

Recently, several cases of adult T-cell leukemia (ATL) with CD30 antigen have been reported, but its clinical significance remains unknown. Accordingly, we studied CD30 antigen expression in ATL cases and documented the clinicopathological characteristics of these cases.

Immunohistochemical and clinical characteristics were studied in 46 patients with malignant lymphoma or benign lesions of lymphoid tissue, who had antibodies against human T-cell leukemia virus type I (HTLV-I). Monoclonal integration of HTLV-I provirus was demonstrated in the tumor cells in 36 (ATL) of the 46 cases. CD30 antigen expression was evident in seven of these 36 cases (19.4%), however it was not seen in any of the ten cases lacking the integration of HTLV-I provirus. A comparison of ATL cases with and without CD30 antigen expression revealed significantly larger numbers of abnormal lymphocytes in the peripheral blood and lower serum calcium levels in ATL expressing CD30 antigen.     

Differences Between Long- and Short-Term Survivors with Lymphoma Type of Adult T-Cell Leukemia

The clinical features at time of diagnosis of long-term survivors with lymphoma type of adult T-cell leukemia (ATL) were compared with those of short-term survivors. We had 51 Japanese patients with lymphoma type of ATL from 1981 to 1989 who had human T-cell leukemia virus type I (HTLV-I) antibody and monoclonal integration of HTLV-,I proviral DNA in the malignant cells. Of the 51 patients, 7 survived for more than 3 years, and they were classified as long-term survivors. Twenty-four patients died within 1 year and they were classified as short-term survivors. Differences between these two groups were investigated with the clinical findings recorded at the time of diagnosis. Findings that proved significant were serum lactate dehydrogenase (LDH) levels, calcium, total protein levels and the presence of B symptoms. Patients with lymphoma type of ATL are expected to be long-term survivors if they have no hypercalcemia or B symptoms with only mildly elevated serum LDH and total protein levels.     

High expression of the longevity gene product SIRT1 and apoptosis induction by sirtinol in adult T-cell leukemia cells

Adult T-cell leukemia-lymphoma (ATL) is an aggressive peripheral T-cell neoplasm that develops after long-term infection with human T-cell leukemia virus (HTLV-1). SIRT1, a nicotinamide adenine dinucleotide(+) -dependent histone/protein deacetylase, plays a crucial role in various physiological processes, such as aging, metabolism, neurogenesis and apoptosis, owing to its ability to deacetylate numerous substrates, such as histone and NF-κB, which is implicated as an exacerbation factor in ATL. Here, we assessed how SIRT1 is regulated in primary ATL cells and leukemic cell lines. SIRT1 expression in ATL patients was significantly higher than that in healthy controls, especially in the acute type. Sirtinol, a SIRT1 inhibitor, induced significant growth inhibition or apoptosis in cells from ATL patients and leukemic cell lines, especially HTLV-1-related cell lines. Sirtinol-induced apoptosis was mediated by activation of the caspase family and degradation of SIRT1 in the nucleus. Furthermore, SIRT1 knockdown by SIRT1-specific small interfering RNA caused apoptosis via activation of caspase-3 and PARP in MT-2 cells, HTLV-1-related cell line. These results suggest that SIRT1 is a crucial antiapoptotic molecule in ATL cells and that SIRT1 inhibitors may be useful therapeutic agents for leukemia, especially in patients with ATL.     

Adenosine deaminase in plasma of patients with adult T-cell leukemia (ATL): correlation between enzyme activity and the ATL subtypes

Adenosine deaminase (ADA) was assayed in plasma from 14 patients with adult T-cell leukemia (ATL) (eight with acute ATL and six with smoldering or chronic ATL), 20 male family members (ten were anti-ATLA antibody positive and the other ten negative), and 25 normal individuals. ADA activity was uniformly higher in plasma from patients with ATL than normal controls. This enzyme activity significantly increased in acute ATL in comparison to smoldering or chronic ATL. In families of ATL patients, no statistical difference in ADA activity between the anti-ATLA antibody-positive group and -negative group could be discerned. The enzyme activity in a patient with acute ATL, after a bone-marrow transplant, rapidly increased as leukemic cells increased in peripheral blood. These findings indicate that the levels of ADA activities in plasma from ATL patients reflect the condition of this disease. Thus, measurement of this enzyme activity offers a further parameter to distinguish subtypes of ATL, and is of prognostic and therapeutic value.     

Coexistence of acute monoblastic leukemia and adult T-cell leukemia: possible association with HTLV-I infection in both cases?

A 64 year-old Japanese man who developed acute monoblastic leukemia during the course of adult T-cell leukemia/lymphoma (ATL) was studied. Leukemic cells in the peripheral blood and bone marrow were monoblasts positive for alpha-naphthol butyrate esterase (alpha-NBE) staining, CD11c and CD36 antigens, whereas tumor cells in the pleural effusion were ATL cells positive for CD2, CD4, CD25, CD29 and CD45RA antigens. These two malignant cells had different chromosomal abnormalities. Monoclonal integration of human T-cell leukemia virus type I (HTLV-I) proviral DNA and T-cell receptor C beta gene (TCR C beta) rearrangement were detected in the ATL cells, but not in the leukemic monoblasts. By polymerase chain reaction (PCR) in the peripheral blood mononuclear cells (CD11c+ 98%, CD2+ 4%, CD20+ 0%) not containing ATL cells, the presence of the gag region of HTLV-I was confirmed. These facts indicate that a double positive T cell (CD29+, CD45RA+) was possibly the target cell for HTLV-I infection and that HTLV-I was not directly related to the oncogenesis of the monocyte lineage in the present case, even if it did infect the monocytes. However, there is still an outside possibility that HTLV-I induced acute monoblastic leukemia indirectly.     

Comparative study of cutaneous T-cell lymphoma and adult T-cell leukemia/lymphoma. Clinical, histopathologic, and immunohistochemical analyses

An important disease entity distinct from cutaneous T-cell lymphoma (CTCL) in Japan is adult T-cell leukemia/lymphoma (ATL), which usually shows the same phenotype as CTCL, i.e., a helper/inducer T-cell phenotype (CD4+CD8-), and usually involves the skin. Clinically, both CTCL and ATL are heterogeneous in nature. In this study, we demonstrated differences between CTCL and ATL in terms of clinical and immunopathologic cell surface features. In patients with ATL, the predominant clinical findings were peripheral lymph node involvement, skin lesions, hepatosplenomegaly, leukemic manifestations, and an aggressive course. In patients with CTCL, by contrast, only skin lesions predominated at the onset of the disease and a relatively good prognosis was demonstrated. Phenotypic heterogeneity of ATL in the skin, i.e., CD4-CD8-, CD4+CD8-, and CD4-CD8+, was demonstrated. Expression of Leu8, CD7 (Leu9), and CD45RA (2H4) was high in both the skin-infiltrating ATL cells and peripheral blood and lymph node ATL cells compared with that in the skin-infiltrating CTCL cells. Expression of CD25 (IL-2R), CD71 (OKT9), HLA-DR, and HLA-DQ was higher in the skin-infiltrating ATL cells than in CTCL cells. Expression of CD29 (4B4) was high, and that of CD45RA (2H4) was low in both the skin-infiltrating ATL and CTCL cells compared with the peripheral blood and lymph node ATL cells. Expression of CD45RO (UCHL-1) was not significantly high in the skin-infiltrating CTCL cells compared with that in ATL cells. The most significant phenotypic difference between ATL cells and CTCL cells was the expression of Leu8 (lymph node homing receptor), CD7 and CD25 antigens on the cell surface, and the main phenotypic difference between skin-infiltrating ATL and CTCL cells and peripheral blood and lymph node ATL cells was the expression of CD29 and CD45RA. These findings confirm that the difference in antigen expression on the cell surface might reflect the clinical features of ATL and CTCL, and suggest that the predominant phenotype of peripheral blood and lymph node ATL cells is that of naive, relatively immature or activated T-cells, and that CTCL cells are previously activated (memory) T-cells. In other words, CTCL cells do not share the same origin as ATL cells. These observations support the concept that ATL is a disease distinct from CTCL.     

Novel Interleukin-2 Dependent T-Cell Line Derived from Adult T-Cell Leukemia Not Associated with Human T-Cell Leukemia Virus Type I

A novel interleukin-2 (IL-2)-dependent T-cell line, WHN2, was established from a patient with adult T-cell leukemia (ATL) not associated with human T-cell leukemia virus type I (HTLV-I). Neither the original leukemic cells nor the WHN2 cells showed proviral integration in their cellular DNAs by Southern blot analysis. The surface phenotype showed that both the original leukemic cells and the WHN2 cells had a common phenotype of ATL, i.e., positive for CD2, CD4, human leukocyte antigen DR (HLA-DR) and CD25, but negative for CD8, a characteristic of helper/inducer T-cells. Most of the chromosomal abnormalities of the original leukemic cells were maintained in the WHN2 cell line. Furthermore, Southern blot analysis of the T-cell receptor β -chain gene rearrangement revealed that the original leukemic cells and WHN2 cell line had identical patterns, suggesting that the WHN2 cell line was truly derived from the original leukemic cells. Dose-dependent growth on IL-2 was demonstrated, and at the maximal stimulation, the number of cells doubled within three days. This IL-2-dependent growth was inhibited by the simultaneous existence of anti-IL-2 receptor a and β chain antibodies. These results indicate that the character of the WHN2 cell line is similar to that of the cell lines derived from ATL associated with HTLV-I. Thus, the HTLV-I-negative ATL cell line, WHN2, should be useful in the comparative study of the pathogenesis of ATL associated with or without HTLV-I.