Search PubMed⌕ Search

Biomedical subjects

S Fukuhara

Publications and source records attributed to S Fukuhara.

At least 217 records · Page 12Linked to original sources

Reed-Sternberg cells in Hodgkin's cell lines HDLM, L-428, and KM-H2 are not actively replicating: lack of bromodeoxyuridine uptake by multinuclear cells in culture.

We compared the proliferation of mononuclear and multinuclear cells in four Hodgkin's cell lines, HDLM-1, HDLM-1d, L-428, and KM-H2, by examining their capacity to incorporate bromodeoxyuridine (BrdUrd) into nuclei. Approximately 5% of all cells in HDLM-1 cultures had two or more nuclei, a characteristic of Reed-Sternberg (RS) cells. Unlike mononuclear Hodgkin's (H) cells, these RS cells exhibited no uptake, or only minimal uptake of BrdUrd, suggesting that they did not replicate actively. Cytogenetic study showed that 25% of the HDLM-1 cells contained a tetraploid (4X) set of chromosomes with a characteristic two-peak distribution. Following treatment of HDLM-1 cells with phorbol ester, the percentages of 4X cells and RS cells increased to 50% and 12%, respectively. This increase in RS cells was not likely to be due to cell fusion as shown by the absence of hybridization of BrdUrd-positive and -negative nuclei. Phorbol ester has a short-term effect of blocking the exit of cells from G1 into S phase, but no effect on the transition from S phase to G2/M phase. The block is more prominent in 2X cells than in 4X cells, which may explain the increase in percentage of 4X cells in phorbol ester-treated cultures. In addition, phorbol ester induced the differentiation of H-RS cells, which was accompanied by loss of the marker HeFi-1 from the cell surface. Approximately one third of the RS cells did not express HeFi-1, or expressed only minimal amounts. The findings led us to the following conclusions: (1) The 4X cells probably are formed from 2X H cells as a result of disturbed cytokinesis, but not a cell fusion. (2) A considerable number of 4X cells were H cells, because the number of 4X cells consistently exceeded that of RS cells. (3) Since mitotic figures are extremely rare in RS cells and these cells did not show active BrdUrd uptake, the increased number of RS cells must also be a consequence of disturbed cytokinesis of H cells or a result of nuclear transformation (twisting, convolution, or separation of the nucleus) in H cells. (4) Most RS cells lose their proliferating capacity and some RS cells may undergo further differentiation. Uptake of BrdUrd and phorbol ester induction were also studied on the other three H-RS cell lines, HDLM-1d, L-428, and KM-H2, with results similar to those for HDLM-1.

Biological Transport↗

Significance of extra 18q- chromosome in Japanese t(14;18)-positive lymphoma.

Karyotype evolution of t(14;18)-positive lymphoma was studied in 13 Japanese patients. The extra 18q- chromosome, found in six of ten patients with complex karyotypes, was the most common change subsequent to a t(14;18)(q32;q21) chromosome translocation. The additional change was interpreted as being a duplication of an 18q- derived from a t(14;18). The six patients had transformed histology of follicular small cleaved cell lymphoma or diffuse large cell lymphoma, and five of them had extranodal expansion associated with a poor prognosis. These findings indicate that the extra 18q-, together with other chromosome abnormalities, is closely associated with the advanced grade disease of t(14;18)-positive lymphoma, and the extra chromosome is evolutionally comparable with the second Philadelphia (Ph1) chromosome often found in the blastic phase of chronic myelocytic leukemia carrying a t(9;22)(q34;q11). In addition, since the extra 18q- is rarely found in American patients with t(14;18)-positive lymphoma, there appears to be a difference in the karyotype evolution between Japanese and American patients.

Chromosome Aberrations↗

c-yes and bcl-2 genes located on 18q21.3 in a follicular lymphoma cell line carrying a t(14;18) chromosomal translocation.

The c-yes-1 and bcl-2 genes have been independently mapped at the same position of the chromosome band, 18q21.3, which is a breakpoint in the t(14;18) chromosomal translocation. High-molecular-weight genomic DNAs and mRNA isolated from a follicular lymphoma cell line FL-18 and an Epstein-Barr virus harboring FL-18EB cell line, which carried the t(14;18), were analyzed. Although there was no detectable c-yes rearrangement, the chromosome 18-specific probe b (bcl-2) revealed one rearranged fragment after EcoRI, BamHI, PstI and SstI digestion. Therefore, breakage of chromosome 18 in the t(14;18) occurred at the SstI fragment containing the probe b sequence on the bcl-2 gene. The transcript of c-yes was undetectable, whereas active transcription of bcl-2 was observed in the FL-18 and FL-18EB cell lines at a higher level than in other types of lymphoma cell line. These data suggest that the c-yes-1 is not involved in the t(14;18), and the 2 genes located at 18q21.3 are independent sequences.

Aged↗

Lymphoma cell line (FL-18) and Epstein-Barr virus-carrying cell line (FL-18-EB) obtained from a patient with follicular lymphoma: monoclonal derivation and different properties.

A novel cell line, FL-18, was established from the pleural effusion of a patient with follicular small cleaved cell lymphoma. At the same time, an Epstein-Barr virus (EBV) nuclear antigen (EBNA)-positive cell line, FL-18-EB, was established from the EBV-infected culture of the same pleural effusion cells. Both cell lines had the same monoclonal surface immunoglobulin (IgG kappa), and they had the same karyotype as that of the fresh pleural effusion cells in which a reciprocal translocation between the long arm of chromosomes 14 and 18 [t(14;18)(q32;q21)] was detected. Gene rearrangement analysis of immunoglobulin heavy-chain gene (JH) and kappa light-chain gene (J kappa) showed the same rearranged configuration in the two cell lines; however, some morphological and phenotypic differences were found. The FL-18-EB cells, which were morphologically similar to common EBNA-positive lymphoblastoid cell lines of normal B cell origin at the initial phase of culture, were larger than the FL-18 cells and contained multinucleated giant cells. The FL-18 cells lacked cytoplasmic immunoglobulin and were positive for common acute lymphoblastic leukemia antigen (CALLA), whereas the FL-18-EB cells had cytoplasmic immunoglobulin and were negative for CALLA. Thus, the phenotype of FL-18-EB seems to be a result of a shift by EBV infection to a more mature stage in the B cell differentiation pathway than that of FL-18. The paired availability of EBV-free and EBV-infected cell lines of a neoplastic clone is unique and valuable in considering EBV infectibility of neoplastic B cells and resultant phenotypic changes.

Aged↗

Establishment of an interleukin 2-dependent human T cell line from a patient with T cell chronic lymphocytic leukemia who is not infected with human T cell leukemia/lymphoma virus.

We established an interleukin 2 (IL-2)-dependent human T cell line, Kit 225, from a patient with T cell chronic lymphocytic leukemia (T-CLL) with OKT3+, -T4+, -T8- phenotype. Southern blot analysis showed that Kit 225 is not infected with human T cell leukemia/lymphoma virus (HTLV) type I or II, and is probably derived from the major clone in the fresh leukemic cells. Kit 225 cells express a large amount of IL 2 receptors constitutively and their growth is absolutely dependent on IL 2. No other stimuli, such as lectins or antigens, are required for maintaining the responsiveness to IL 2. As abnormal IL 2 receptor expression was also seen originally in the fresh leukemic cells, the establishment of this cell line with IL 2 suggests that IL 2-mediated T cell proliferation is involved in the leukemogenesis of some cases of HTLV-negative T-CLL.

Antigens, Surface↗

Establishment of a new myeloid leukemia cell line (TK-1), and isolation of cells having a translocation involving a band 17q23.

A myeloid leukemia cell line (TK-1) was established from the peripheral blood of a patient with lymphoblastic lymphoma whose leukemia cells were composed of T-lymphoblasts and immature myeloid cells. The established TK-1 cell line consisted of immature myeloid cells with heavy azurophilic granulation in the cytoplasm. The TK-1 cells were positive for peroxidase, and exhibited a strong positive reaction for alpha-naphthyl acetate esterase and naphthol AS-D chloroacetate esterase. The cells were weakly positive for Fc gamma-receptors, showed no phagocytosis and did not reduce NBT. With the treatment of 1 alpha, 25-dihydroxy-vitamin D3, they exhibited morphological and functional differentiation. The TK-1 cell line contained normal diploid cells and pseudodiploid cells, and the two populations were successfully cloned; the clone with a normal karyotype was designated the TK-1D cell line, and the clone with a pseudodiploid karyotype, which had a translocation involving chromosomes 14, 17 and one other chromosome, was designated the TK-1B cell line. These cloned cells lacked Epstein-Barr virus nuclear antigens and had almost the same myelomonocytic characteristics as the parent TK-1 cells. The breakpoint of chromosome 17 involved in the translocation of the pseudodiploid cells was identified to be a band 17q23.

Adult↗

Recurrent chromosome abnormalities in adult T-cell lymphomas of peripheral T-cell origin.

Cytogenetic studies were performed on 11 Japanese patients with adult T-cell lymphoma of peripheral T-cell origin. All patients had a histologic diagnosis of diffuse lymphoma, and were treated with intensive combination chemotherapy; their median survival was 6 months. Lymphomas were categorized on the basis of the classification proposed by the Lymphoma Study Group of Japan: 5 patients had the features of pleomorphic lymphoma; 3, mixed lymphoma; 2, large-cell lymphoma; and 1, medium-sized-cell lymphoma. The modal chromosome number of abnormal cells was near-diploid in 7, and hypotetraploid in 4 including 3 patients with pleomorphic lymphoma. Abnormalities of the clonal chromosomes were observed in all 11 patients. A translocation of the short arm of 19 at band 19p13 was found in 2 patients with pleomorphic lymphoma and in one patient each with mixed lymphoma and medium-sized-cell lymphoma. A translocation of the short arm of 14, with a break at band 14p12, occurred in 4 patients with pleomorphic lymphoma. In one patient each with mixed lymphoma and large-cell lymphoma, an extra chromosome 3 was found. The numerical change was unique because their lymphomas were further categorized as "angioimmunoblastic lymphadenopathy with dysproteinemia" and "large-cell immunoblastic lymphoma", respectively. Our findings suggest that characteristic chromosome abnormalities occur in adult T-cell lymphoma, and dividing cells in the tetraploid range tend to be predominant in pleomorphic lymphoma.

Adult↗

Cytochemical, immunologic, chromosomal, and molecular genetic analysis of a novel cell line derived from Hodgkin's disease.

A novel cell line, KM-H2, was established from the pleural effusion of a patient with Hodgkin's disease of mixed cellular type. Multiple phenotypic studies were carried out with this cell line. Acid phosphatase and nonspecific esterase activities were detected. Rosette formation with T lymphocytes and the receptors for C3b and Fc portion of IgG were positive. Among the antigens tested with a total of 22 monoclonal antibodies defining hematopoietic cell subsets or lineages, Ki-1, Leu-M1, MCS1, HLA-DR, and OKT9 antigens were found to be positive. The other antigens reportedly specific for T cells, B cells, natural killer (NK) cells, monocytes, interdigitating reticulum (IR) cells and dendritic reticulum cells were negative. These phenotypic features were identical to those of the Sternberg-Reed (SR) and Hodgkin (H) cells in the fresh materials reported by other researchers. Moreover, the KM-H2 cells and the parental pleural effusion cells shared several structural chromosome anomalies. These findings indicated that the KM-H2 cells are derived from the SR and H cells. Molecular genetic analysis of the KM-H2 cells disclosed that the human immunoglobulin JH gene was rearranged but not the JK gene, and that the human T cell receptor beta chain gene was of the germline type. Based on these properties of the KM-H2 cells, Hodgkin's disease may be derived from a cell lineage other than T cell or B cell.

Adult↗

Establishment of a novel acute monoblastic leukemia cell line (YK-M2) having a near-triploid karyotype.

The YK-M2 cell line was established from the peripheral blood of a patient with acute monoblastic leukemia in whom an anterior mediastinal tumor preceded the peripheral blood manifestation. The established cells grew in a single cell suspension with a doubling time of 60 h and consisted of primitive monoblastic cells. The cells were 52% positive for peroxidase staining and manifested strongly positive activity of alpha-naphthyl acetate esterase, which was completely inhibited by sodium fluoride. The cells showed strong expression of Fc gamma receptors and phagocytosed sensitized ox erythrocytes. When the cells were incubated with 1 alpha,25-dihydroxy-vitamin D3, they were induced to differentiate into mature monocyte-macrophage-like cells, which reduced the nitroblue tetrazolium dye and released a small amount of the superoxide anion. Cytogenetic studies revealed that the cells had a near-triploid karyotype with a modal chromosome number of 68, and the short arm of one No. 17 chromosome was deleted [del(17)(p11)]. The YK-M2 cell line is particularly unique in that the cells retained the polyploid karyotype that may be an initial cytogenetic change in the malignant transformation of the parent leukemia cells.

Adult↗

Phenotypic analysis of acute lymphoblastic leukemia (ALL) cells which are classified as non-T non-B and negative for common ALL antigen.

When phenotypic marker analysis of acute lymphoblastic leukemia (ALL) cells (102 cases) was performed, a group of ALL cells (15 cases) classified as non-T non-B, and negative for common-ALL antigen (CALLA) was characterized in a focused manner. "Non-T non-B" was defined as negative for T cell properties such as E-rosetting or reactivity with anti-human T-cell monoclonal antibodies (T101, WT1), and absence of any B-cell characteristics (cell surface and/or cytoplasmic immunoglobulin and reactivity with B1 monoclonal antibody). Despite their marked heterogeneity, CALLA(-) non-T non-B ALL cells revealed three different phenotypic patterns in terms of presence of terminal deoxynucleotidyl transferase (TdT) and of reactivity with antimyeloid (MCS1) or myelomonocyte (MCS2 and OKM1) monoclonal antibodies. Four of 15 cases reacted with some myeloid-specific antibodies, but were negative for TdT. Six cases had both MCS2 antigen and TdT. The remaining five cases expressed no myeloid antigens, but were positive for TdT with some exceptions. These findings showed that acute leukemias with myeloid antigens might be involved in CALLA(-) non-T non-B ALL having no relationship to the presence of TdT, and, furthermore, that the blasts with simultaneous expression of TdT and myeloid-specific antigen (MCS2) might represent an immature stage in hematopoietic differentiation closely corresponding with the bifurcation of the lymphocyte/myeloid pathway. Alternatively, only five cases remained "unclassified leukemia." We therefore think that the detailed examination of CALLA(-) non-T non-B ALL cells using myeloid specific antibodies is helpful in clarifying the characteristics of myeloid precursors and the common bipotential stem cell of lymphoid and myeloid progenitors.

Adult↗