Isao Sanada

Polyclonal integration of HTLV-I proviral DNA in lymphocytes from HTLV-I seropositive individuals: an intermediate state between the healthy carrier state and smouldering ATL

We have studied 15 individuals (aged 14‐74 years) with antibodies to HTLV‐I in their serum and random integration of HTLV‐I proviral DNA in their peripheral blood lymphocytes. All but one of these patients suffered from a variety of non‐specific complaints which did not correspond to those of adult T‐cell leukaemia (ATL). All of them were born in Kyushu and Okinawa which are endemic areas for HTLV‐I infection; 25% of their family members were also seropositive for HTLV‐I. The only haematological abnormality in these patients was the presence of few atypical lymphoid cells in the peripheral blood. The CD4/CD8 ratios were normal but the proportion of Tac positive cells was slightly higher than normal.

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.

Treatment of post-transplanted, relapsed patients with hematological malignancies by infusion of HLA-matched, allogeneic-dendritic cells (DCs) pulsed with irradiated tumor cells and primed T cells.

Patients with hematological malignancies who relapse after bone marrow transplantation (BMT) are often treated with donor lymphocyte infusion. However, this procedure often results in graft-versus-host disease (GVHD). While, Dendritic cells (DCs), which present antigens to naive T cells, have been used in the immunotherapy of cancer, this approach has been logistically difficult due to limiting numbers of DCs. We have now developed a method for obtaining a large number of DCs by treating the granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood stem cells (PBSCs) from healthy donors with granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4), and tumor necrosis factor-α (TNF-α). The resulting cells possess the morphologic, phenotypic, and functional characteristics of mature DCs. In in vitro studies, culture of these HLA-matched donor derived-DCs with irradiated each patients tumor cells as an antigen source, followed by incubation with T cells from the patient, induced the production of highly cytotoxic T lymphocytes (CTLs) specific for the respective tumor cells in the semi-allogeneic setting. A transient, but objective clinical response was obtained in the absence of GVHD when we injected the DCs which had been pulsed with irradiated tumor cells as well as primed T cells from the same original donor of related-allogeneic stem cell transplantation into the relapsed patients. Our findings suggest that treatment of relapsed patients with such donor-derived DCs, and primed T cells may be effective as an adjunctive immunotherapy.

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.

Trisomy 4 in a case of acute myelogenous leukemia accompanied by subcutaneous soft tissue tumors : report of a case and review of the literature

We report a patient with acute myelogenous leukemia [AML, French-American-British classification (FAB) M2] with trisomy 4, who developed subcutaneous soft tissue tumors at the time leukemia was diagnosed. A review of the literature on AML with trisomy 4 suggests a relation between trisomy 4 and tumor formation of leukemic cells.

Amylase-Producing Multiple Myeloma

Hyperamylasemia resulting from ectopic production of amylase by tumor tissue was first described by Weiss et al. (1951). Since then, several cancer cases of hyperamylasemia have been reported, including lung (Maeda et al. 1982; Katayama et al. 1981; Braganza et al. 1978; Yokoyama et al. 1977), ovary (Hodes et al. 1985; Cramer et al. 1979; Sandiford et al. 1978; Corlette et al. 1978), stomach (Nomura et al. 1980), and others (Yoshimura et al. 1986; Ueda et al. 1977; Matsuyama et al. 1979; Weitzel et al. 1988; Maesiar 1984). Most of these have been derived from epithelial tissues. To our knowledge, we reported the first case of an amylase-producing multiple myeloma (Hata et al. 1988). We also established an amylase-producing multiple myeloma cell line, KHM-1B from the patient (Matsuzaki et al. 1988).

Chromosomal abnormalities in a patient with smoldering adult T-cell leukemia: evidence for a multistep pathogenesis

A cytogenetic study was performed on peripheral blood cells from a patient with smoldering adult T-cell leukemia (ATL). Four types of primary abnormal clones were found upon examination of a large number of karyotypically analysed cells cultured with and without phytohemagglutinin (PHA). However, human T-cell leukemia virus (HTLV) proviral DNA was confirmed to be monoclonal. This discrepancy can be explained by the hypothesis that these four primary abnormal clones were all derived from a leukemic clone with a normal karyotype and the same integration site of HTLV proviral DNA.

Human T‐Cell Leukemia Virus‐1‐positive Cell Line Established from a Patient with Small Cell Lung Cancer

A stable cell line, KHM‐3S, was established from a patient with small cell lung cancer (SCLC), who had a high serum level of soluble interleukin 2 receptors (sIL2‐R) and was seropositive for human T cell leukemia virus (HTLV)‐l. KHM‐3S cells were positive for IL2‐R (Tac) and NKH‐1, but negative for other lymphocytic markers such as OKT 11, OKT 4, OKT 8, T cell receptor (WT 31), B 1, and B 4. Moreover, the KHM‐3S cells were negative for leukocyte common antigen and strongly positive for neuron‐specific enolase (NSE). Secretion of sIL2‐R and NSE by the KHM‐3S line was detected by an enzyme‐linked immunosorbent assay. Rearrangement of the T cell receptor gene and monoclonal HTLV‐1 integration were found by Southern blot analysis of KHM‐3S DNA. However, Northern blot analysis showed no T cell receptor mRNA. KHM‐3S may be useful for studies on the role of HTLV‐1 in carcinogenesis and IL2‐R expression in SCLC.

Establishment of a human T-cell line with deficient surface expression of glycosylphosphatidylinositol (GPI)-anchored proteins from a patient with paroxysmal nocturnal haemoglobinuria.

A novel interleukin‐2 dependent T‐cell line, PMT‐2Y, was established from the peripheral blood of a patient with paroxysmal nocturnal haemoglobinuria (PNH) by human T lymphotropic virus type I (HTLV‐1)‐mediated transformation. PMT‐2Y cells are positive for CD2, CD3, CD4, CD25, T cell receptor αβ and HLA‐DR, but negative for CD1, CD7, CD8, CD19 and CD20, indicating that the clone belongs to a helper/inducer subset of T cells. PMT‐2Y cells have the monoclonal integration of HTLV‐I proviral DNA, suggesting that they derived from a single clone. Moreover, they lack surface expression of complement regulatory proteins such as DAF (CD55) and CD59, that are the most important glycosylphosphatidylinositol (GPI)‐anchored membrane proteins defective in haemopoietic cells of patients with PNH. Northern blot analysis, however, revealed the production of normal levels of DAF mRNAs. Thus, PMT‐2Y is derived from a PNH T cell clone and may be a useful model to study PNH.

Genetic predisposition to the Crow-Fukase syndrome

Histocompatibility antigens (HLA-A,B,C,DR, and DQ antigens) were investigated in 25 Japanese patients with the Crow-Fukase syndrome (Takatsukis disease). No significant associations were detected between the patients with the Crow-Fukase syndrome and the healthy controls.