Kiyoshi Takatsuki

A new G-CSF-supported combination chemotherapy, LSG15, for adult T-cell leukaemia-lymphoma : Japan Clinical Oncology Group Study 9303

This phase II trial was performed to evaluate the efficacy of a new granulocyte colony‐stimulating factor (G‐CSF)‐supported multi‐agent chemotherapy protocol, LSG15, for aggressive adult T‐cell leukaemia‐lymphoma (ATL). Ninety‐six previously untreated patients with aggressive ATL were enrolled and grouped as: acute type (58), lymphoma type (28) and unfavourable chronic type (10). Therapy consisted of seven cycles of VCAP (vincristine, cyclophosphamide, doxorubicin and prednisone), AMP (doxorubicin, ranimustine and prednisone) and VECP (vindesine, etoposide, carboplatin and prednisone). G‐CSF was administered during the intervals between chemotherapy until neutrophil reconstitution was achieved. Eighty‐one per cent of the 93 eligible patients responded [95% confidence interval (CI), 71·1–88·1%], with 33 patients obtaining complete response (35·5%) and 42 obtaining partial response (45·2%). The median survival time (MST) after registration was 13 months and the median follow‐up duration of the 20 surviving patients was 4·2 years (range 2·8–5·6). Overall survival at 2 years was estimated to be 31·3% (95% CI, 22·0–40·5%). Grade 4 haematological toxicity of neutropenia and thrombocytopenia were observed in 65·3% and 52·6% of the patients respectively, but grade 4 non‐haematological toxicity was observed in only one patient. LSG15 is feasible with mild non‐haematological toxicity and improved the clinical outcome of ATL patients. MST and overall survival at 2 years were superior to those obtained by our previous trials.

Effect of adult T-cell leukemia cells on pokeweed mitogen-induced normal B-cell differentiation

Abstract Leukemic T cells from Japanese patients with adult T-cell leukemia were studied for their effects on pokeweed mitogen-induced peripheral blood lymphocyte differentiation into immunoglobulin-producing cells. Allogeneic peripheral blood lymphocytes, normal T cells, nonlymphoid leukemia cells, lymphoid leukemia cells other than adult T-cell leukemia cells, and long-term T-cell lines did not suppress normal B-cell differentiation in coculture experiments. Leukemic T cells from three of six patients with adult T-cell leukemia showed a marked suppressive effect. Supernatant fluids obtained from a short-term (48-hr) culture of leukemic T cells from one of these three patients also showed a marked suppressive effect on B-cell differentiation at a final dilution of 1 5 or less when added at the beginning of the culture (on Day 0) or on Day 1. Supernatant fluids, however, did not manifest suppressor activity when added on Day 3 or later, or when heated at 56°C for 30 min before addition.

Uveitis Associated With Human T-cell Lymphotropic Virus Type I

Seroepidemiologic, clinical, and virologic studies were performed to determine whether human T-cell lymphotropic virus type I was closely associated with uveitis in two hospitals. One hospital was in an endemic area of the virus (Miyakonojo, Miyazaki) and the other hospital was in a less endemic area (Kurume). In the endemic area, the seroprevalence of the virus in patients with uveitis without defined causes (35.4%, 62 of 175 patients) was significantly higher than that in patients with nonuveitic ocular diseases (16.1%, 42 of 261 patients), or in patients with uveitis with defined causes (10.3%, eight of 78 patients). The seroprevalence in younger patients (20 to 49 years of age) with uveitis without defined causes in the area was 44.8% (30 of 67 patients), whereas it was only 9.3% (ten of 107 patients) in the other two groups. A similar observation was recorded even in the less endemic area (Kurume). Because the seroprevalence of the virus in the general population is known to be low in younger patients and to increase with age, these findings were interpreted to indicate that the association of human T-cell lymphotropic virus type I with uveitis was significant. Most patients, particularly those aged 20 through 49 years, had an intermediate uveitis characterized by a moderate inflammation in the vitreous body accompanied by an iritis and retinal vasculitis. The ocular symptoms in the patients differed from those of other types of uveitis common in Japan (Behçets disease, Vogt-Koyanagi-Haradas disease, and toxoplasmosis, for example).(ABSTRACT TRUNCATED AT 250 WORDS)

HTLV-I seroprevalence in patients with malignancy

Since many malignancies often occur in patients with smoldering type adult T‐cell leukemia (ATL) (5 of 18 cases in this report), the relationship between HTLV‐I (human T‐cell leukemia virus type I) infection, which is closely associated with ATL, with other malignancies in an HTLV‐I endemic area was examined. Among the 394 patients with malignancies and who had not had blood cell transfusions, 61 (15.5%) tested positive for HTLV‐I antibody. The prevalence was significantly higher in males older than age 40 years and females of all ages compared to age‐and sex‐matched healthy individuals. The overall seroprevalence (26.1%) in 291 patients with malignancies and who had had blood cell transfusions was higher than that of those who had not had blood transfusions. There was no significant correlation between the site of malignancy and antibody prevalence. These results suggest the possibility that development of malignancy may contribute to expression of latent HTLV‐I infection and that HTLV‐I infection may contribute to the risk of other malignancies.

Familial adult T-cell leukemia.

Two siblings who developed adult T‐cell leukemia (ATL) are presented. The patient and 7 of 26 healthy family members examined had the serum antibodies against ATL‐associated antigens (ATLA). This family study shows that two main routes of transmission of human T‐cell leukemia virus (HTLV) may be involved: one is the route from parents to children and the other is horizontal transmission among spouses, especially from husband to wife; the anti‐ATLA‐positive family is considered to be a high‐risk group for ATL.

Synthesis of a novel cytokine and its gene (LD78) expressions in hematopoietic fresh tumor cells and cell lines

The gene coding for the protein LD78 was isolated from stimulated human tonsillar lymphocytes by differential hybridization. The gene product consisted of 92 amino acids with characteristics of cytokines. LD78 gene transcripts were detected in eight of eight fresh samples of cells from patients with acute nonlymphocytic leukemia (ANLL) by Northern blot analysis. ANLL cells with monocytic features gave the strongest bands. RNA transcripts were found in two of three samples of cells from patients with adult T cell leukemia (ATL), eight of nine samples from patients with acute lymphocytic leukemia (ALL) of B cell lineage, and one of the three samples from patients with T cell ALL. KG-1, HL-60, HUT 102, MT-2, and MJ cell lines expressed the LD78 gene constitutively. The LD78 protein was detected in culture supernatants and cell lysates of HUT 102, MT-2, MJ, and fresh ATL cells by Western blot analysis. This protein was not found in culture supernatants or cell lysates of monocytic leukemia cells and HL-60 cells, although LD78 transcripts were found in those cells. The discrepancy between gene and protein expression might be explained by the stability of the mRNA. Thus, the protein may be involved in the neoplastic transformation of hematopoietic cells.

Autocrine growth by two cytokines, interleukin-6 and tumor necrosis factor alpha, in the myeloma cell line KHM-1A

The effects of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) on the cell growth of the myeloma cell line KHM-1A were studied. TNF-alpha strongly induced KHM-1A proliferation, while IL-6 weakly enhanced growth only at low cell densities. When TNF-alpha was combined with IL-6, TNF-alpha-induced growth enhancement was reduced. According to Northern blotting analysis, the m-RNA of both TNF-alpha and IL-6 were detected in KHM-1A. Moreover, monoclonal antibody capable of neutralizing the cytotoxic activity of TNF-alpha inhibited the proliferation of this cell line. These findings suggest that this cell line operates under an autocrine growth mechanism with respect to these two cytokines, IL-6 and TNF-alpha.

Inversion of chromosome 16 and bone marrow eosinophilia in a myelomonocytic transformation of chronic myeloid leukemia.

We report a case of chronic myeloid leukemia (CML) in myelomonocytic transformation associated with bone marrow (BM) eosinophilia. At diagnosis, all BM cells showed a Ph chromosome. At the time of blastic phase, more than 50% of Ph+ cells had a pericentric inversion of chromosome 16, inv(16)(p13q22). This case confirms that blastic transformation of CML can involve any committed progenitor, and myelomonocytic leukemia with BM eosinophilia is specifically associated with rearrangement of chromosome 16 at band p13 and q22.

Interleukin‐2 Prevents Programmed Cell Death in Adult T‐Cell Leukemia Cells

Adult T‐cell leukemia (ATL) is a prototype of the lymphoma/leukemia syndromes involving immunologically mature T‐lymphocytes. The first retrovirus described in humans, HTLV‐I, is causally related to the disease. In this study, we examined whether ATL cells die in vitro through programmed cell death (PCD), which has been shown to occur in cells affected by several other acute and chronic Icukcmias. When ATL cells from peripheral blood were cultured in serum‐free complete medium, a substantial proportion of them spontaneously died by PCD. After 48 h of culture, approximately 30% of the total DNA was fragmented. Electrophoresis indicated that the DNA of the ATL cells had been cleaved into regular oligonucleosome fragments each comprising approximately 180–200 base pairs. This process was significantly promoted by methylprednisolone and the protein kinase A (PKA) activator Sp‐cAMPS in at least some cases. Since all ATL cells possess interleukin‐2 receptors on the cell membrane, the effect of IL‐2 on spontaneous PCD was assessed. PCD after 48 h of culture was inhibited by 30–50% by 100 U/ml interleukin‐2 (IL‐2). This effect of IL‐2 to prevent spontaneous PCD was dose‐ and time‐dependent. These findings suggest that the viability of ATL cells in vivo is regulated positively and negatively by intrinsic IL‐2, glucocorticoid and regulators of PKA activity. Furthermore, the process of cell death may be involved in the development of the disease.

Establishment and characterization of acute B-cell lymphocytic leukemia cell line showing (8;14) and (14;18) chromosome translocation

Cell line KHM-2B expressing two oncogene products, c-myc and bcl-2, was established from a patient with acute lymphocytic leukemia with an 8;14 and 14;18 chromosome translocation. Surface marker studies of the cell line showed that the cells were positive for HLA-DR, CALLA (CD10), B1 (CD20) and B4 (CD19), but negative for T11 (CD2). The fresh cells from peripheral blood of the patient had no surface immunoglobulins, whereas KHM-2B cells were positive for mu.lambda type surface immunoglobulin. A cytogenetic analysis of the cell line revealed two translocations, t (8;14) (q24;q32) and t(14;18)(q32;q21). Rearrangement of the c-myc and bcl-2 genes was detected by Southern blot analysis of the KHM-2B DNA. Northern blot analysis revealed production of c-myc and bcl-2 mRNAs. These results indicated that two oncogenes were activated by two translocations to immunoglobulin genes.