Correlation between karyotype and prognosis in adults with lymphocytic leukemia are reviewed. Concerning acute lymphoblastic leukemia (ALL), patients with normal karyotype or with hyperdiploidy (greater than 50 chromosomes) show good prognosis. On the other hand, patients with structural abnormalities, such as t(8;14), t(4;11), t(11;14) or with hyperdiploidy (47 to 50 chromosomes), show poor prognosis. Above findings indicate that karyotype is a significant prognostic factor in adults as well as in children with ALL. Karyotype must be considered in designing clinical trials to improve the prognosis in patients with lymphocytic leukemia, especially in ALL.

Ph1-positive leukemias consist of acute leukemia (Ph1 AL) and CML. Cytogenetically, Ph1 AL is often associated with +6, -7, +8, +21, or +Ph1. CML is predominantly accompanied by +Ph1, +8, i (17q), +19 in myeloid crisis and +Ph1, +8, +21 in lymphoid crisis. Thus, i(17q) seems specific for myeloid crisis of CML. Ph1 constricts ABL/BCR within M-BCR in CML and in one half of the adult Ph1 AL. BCR breaks upstream to M-BCR in the other half of adult AL and in most of childhood AL. However, the breakpoint does not affect clinical and hematological features in AL. Consequently, there seems to be two types of Ph1 leukemia; one is AL representing m-BCR rearrangement and the other is CML and Ph1 AL showing M-BCR rearrangement.

In 1960, Nowell and Hungerford found, for the first time, a minute chromosome at the metaphase in chronic myelocytic leukemia (CML) cells, which was called Philadelphia chromosome (9; 22 translocation) later. Ph1 chromosome was considered to be specific for the disease and was frequently used as an important marker for the definite diagnosis. In 1970s banding techniques revealed some other specific chromosome abnormalities, like 8; 14, 8; 21, and 15; 17 translocations, for acute leukemias. In 1980s, molecular-biology techniques were applied in the fields of leukemia research. As a result, many break point cluster regions were discovered in relation to the immunoglobulin chain genes and T cell receptor genes (Table 2). In this review, the specificity of chromosome abnormalities as well as genetic changes in types of leukemia is discussed.

Immunophenotypic and immunogenotypic changes in 23 patients with Ph positive chronic myelogenous leukemia in blast crisis were determined using a panel of monoclonal antibodies and gene probes. According to the immunophenotypes, 9 patients were considered to be in lymphoid blast crisis, including 7 patients with lymphoblastic crisis and 2 patients with lymphoid/myeloid mixed blast crisis. Leukemia cells from the remaining 14 patients showed myeloid phenotypes and 10 of these had platelet-associated antigens. Rearrangement of the immunoglobulin (Ig) gene was observed in all the 9 patients with lymphoid blast crisis, and Ig gene rearrangement was associated with the expression of CD19 antigen. Two patients with myeloid blast crisis showed rearrangements of T-cell-receptor gene, but, dissociation between phenotypes and genotypes was not frequently observed in patients with blast crisis.

Phenotypes of leukemic cells can be determined through dual staining with pairs of FITC-labeled and PE-labeled monoclonal antibodies using a laser flow cytometer. Hybrid acute leukemia (HAL) was diagnosed when leukemic cells expressed 2 or more lymphoid markers and at least one myeloid maker simultaneously. Based on this criteria, forty out of 111 cases with untreated acute leukemia were diagnosed as HAL, 16 of 41 (39%) patients with acute lymphoblastic leukemia and 15 of 70 (21%) patients with acute non-lymphocytic leukemia were diagnosed as having HAL. We classified these HAL cells into 4 types by the following items. Type I; leukemic cells expressed only CD33 and 2 or more B-cell antigens, type II; only CD33 and 2 or more T-cell antigens, type III; CD13 and/or CD33 and 2 or more B-cell antigens, and type IV; CD13 and/or CD33 and 2 or more T-cell antigens. There were 7 type I cases and 16 type III cases. The leukemic cells of these two types were thought to be ALL cells expressing one or several myeloid lineage-associated antigens. There were 4 type II cases and 10 type IV cases. Type II and IV were regarded as lymphoid-lineage associated antigens positive AMLs. At least in adults, the expression of myeloid-associated antigens of ALL cells seems to identify a high risk group of ALL patients with a poorer response to standard ALL therapy.

The diagnosis of acute undifferentiated leukemia (AUL) is made when the leukemic cells do not have cytologic or cytochemical features of myeloid cells, and do not express myeloid antigens (CD13, CD14, CD33, CD41 etc.) or lymphoid antigens (CD2, CD3, CD19, CD20, Sm Ig etc.). Most of these cells are reported to be positive for CD7, CD34, TdT and HLA-DR, either alone or in combination. Cell lineage can be suspected in most AUL cells by genotypic analysis, phenotypic analysis after culturing with TPA, or peroxidase activity by ultrastructural or immunohistochemical analysis, which indicates the heterogeneity of AUL. The patients with AUL appear to have a poor prognosis with conventional chemotherapy.

We investigated the surface markers, cell-function, clonality, and the association of IL-2 receptors and a second messenger of src family of tyrosine kinase p56lck in IL-2 signal transduction of the leukemic cells of 12 patients with large granular lymphocytic leukemia (LGL leukemia). The leukemic cells of 5 patients were CD3+ and 5 of them were CD3-. In three patients with CD3- leukemia examined, one showed karyotype abnormality of 46, XY, -10, +mar and the delta gene of TCR was rearranged in one patient. The TCR of the leukemic cells of a patient MH with CD3+, CD4 and CD8 (double positive marker: DP) recognised rabbit IgG presented by macrophages. The recognition was class II restricted. We examined the expression pattern of CD8 subunits and found that DP leukemic cells commonly expressed CD8 alpha+ beta-. These results suggested that DP leukemic cells were CD4+ T cells and expressed CD8 alpha secondarily. The p75 IL-2 receptors were detected, however, the modulation of p56lck in the process of IL-2 signal transduction were not found out. There was no association between p75 and p56lck when leukemic LGL cells proliferated on stimulation with IL-2.

Modern diagnosis and classification of leukemia are reviewed. The FAB (French, American, British) classification, introduced in the late 1970s has been the basis for most studies to date. During the 15 years since then, new categories such as M7 and M0 were added to the classification. The MIC proposal (morphology, immunology, cytogenetics) has been an important development which emerged from the knowledge about chromosomal changes and immunophenotyping. Improvement in diagnosis and classification will emerge from studies employing all the above techniques, including DNA analysis, in the 1990s.

Immunoglobulin heavy-chain (IgH) gene expression is regulated in a large part of the IgH gene intronic enhancer which is composed of certain protein-binding motifs. These motifs act as a DNA element which is important for the regulation of the IgH gene transcription. Two of these motifs, HE2 and E6 in the human IgH gene intronic enhancer, are considered to affect the B cell-specific gene expression. To examine the true function of the HE2 and E6 motifs in vivo, we established transgenic mouse lines. A 3'deletion mutant of the IgH gene intronic enhancer which includes an active HE2 motif showed almost the same degree of transcriptional activity and tissue-specificity as the whole IgH gene intronic enhancer. Still more, using the luciferase-assay, it was confirmed at cellular level that the HE2 motif plays an important role in the tissue-specific activity of the IgH gene intronic enhancer. On the other hand, the E6 motif was unable to sufficiently produce the B cell-specific expression in vivo and showed enhancer activity in some non-B cells. In transgenic mice, the IgH gene intronic enhancer functioned not only in B-lymphocytes but also in chorid plexus cells which are similar to glial cells. This suggests the existence of common gene expression control mechanisms between the immune system and the nervous system.

The present study was designed to determine the effects of follicular stimulating hormone (FSH) on the proliferation of the human ovarian cancer cell line (HRA line) in vitro and in vivo. The results showed: 1) The number of cells and 3H-thymidine uptake significantly increased after FSH treatment, but were suppressed by Buserelin (number of cells: 2.5 +/- 0.4 [con] vs 13.8 +/- 1.7 [FSH] vs 3.0 +/- 0.2 [FSH+GnRHa] x 10(4)/ml, uptake: 1,272.0 +/- 51.5 [con] vs 4,183.4 +/- 114.1 [FSH] vs 885.0 +/- 177.0 [FSH+GnRHa]cpm/10(5) variable cell mean +/- SD, p less than 0.05). 2) FSH increased the proportion of the cells in the S phase but decreased the cells in G0/G1 phase (S: 42.1 +/- 0.72 [con] vs 61.7 +/- 0.5 [FSH], G0/G1: 53.8 +/- 0.4 [con] vs 35.5 +/- 0.6 [FSH]% mean +/- SD, p less than 0.01, t-test). 3) Cyclic adenosine monophosphate (cAMP) production by HRA cells was significantly increased by cholera toxin (CTX), but not by FSH (1.29 +/- 0.72 [con] vs 2.05 +/- 0.21 [FSH] vs 16.2 +/- 2.28 [CTX]nM mean +/- SD p less than 0.001), suggesting that FSH has no effects on the adenylate cyclase system in the cell line. 4) FSH and GnRHa receptors were identified in HRA cells, and the number of the receptors was significantly decreased by Buserelin treatment (426.0 +/- 6.8 vs 98.6 +/- 12.3 sites/cell mean +/- SD, p less than 0.05, t-test).(ABSTRACT TRUNCATED AT 250 WORDS)

PCR-SSCP analysis is a rapid, simple and sensitive technique for detection of various mutations, including single nucleotide substitutions, insertions and deletions, in PCR-amplified DNA fragments. Here I review, the principle and sensitivity of the technique, and also show how this technique has been used in clinical and basic medical sciences.

Colonic cancer cell strain KE43 was established from a human colonic cancer diagnosed histologically as a predominantly well differentiated adenocarcinoma with minute foci of poorly differentiated adenocarcinoma. The well differentiated adenocarcinoma cell line was identified as the major morphological picture in xenografts of KE43 and 58 in nude mice, but this changed to poorly differentiated adenocarcinoma in passage 105. Doubling time of this cancer cell line was 22.5 hours in passage 105. The modal numbers of chromosomes were 41 and 76. Cancer cells could be heterotransplanted in 100% of the nude mice. The tumor cells produced and secreted CA19-9, CEA and Laminin into the spent medium. This cell line appears to provide a useful system for studying colonic cancer in vivo and in vitro.

Human myeloid leukemia cells do not differentiate into functional end-cells but remain in the proliferation pool. Human cell lines can serve as model for hematopoietic cells arrested at different stages of myeloid differentiation and helps to dissect the cellular and molecular events involved in leukemogenesis. Furthermore, several agents have been identified as inducers of differentiation of leukemia cells. Exciting new clinical observation have shown that patients with APL respond well to the treatment with all-trans retinoic acid. RAR-alpha gene was proved to translocated from chromosome 17 to a locus PML on chromosome 15. This new chimeric gene, PML-RAR alpha is extremely important to understand the leukemogenesis of APL.

Expression of multidrug resistance (mdr 1) gene, which encodes a transmembrane efflux pump referred to P-glycoprotein, leads to the decreased intracellular accumulation of various lipophilic drugs, such as vinca alkaloids, anthracyclines and epipodophyllotoxins. As these drugs are commonly used in chemotherapy for acute leukemia, it is of importance to determine whether mdr 1/P-glycoprotein expression is associated with clinical resistance. In several reports, some leukemia cells from untreated patients have expression of mdr 1/P-glycoprotein. We quantitatively detected low levels of mdr 1 expression in all cases of untreated acute leukemia and normal hematopoietic cells, using the reverse transcriptase-polymerase chain reaction. Carefully designed clinical trials including mdr 1 reversing agents may have significant consequences for the treatment of acute leukemia.

The t(1;19)(q23;p13) seen in approximately 5% of childhood acute lymphoblastic leukemia (ALL) has been reported to be associated with leukoencephalopathy. 1;19 translocation can alter the E2A gene, leading to formation of a chimeric E2A-PBX1 gene that retain the activator domain of the E2A gene but substitute a homeobox domain of the PBX1 gene for the helix-loop-helix DNA binding and dimerization domain of E2A. The translocation breakpoints occurs within a single intron of the E2A gene on chromosome 19, and interrupts a homeobox gene, PBX1, on chromosome 1q23. Most cases with t(1;19) have been identified to have rearranged band of E2A by Southern blotting analysis, and to contain identical E2A-PBX1 chimeric transcripts by use of polymerase chain reaction assay. The molecular breakpoints in pre-B cases differ from those in early pre-B cases among t(1;19)-ALL. Thus, molecular analysis is useful for detection of t(1;19)-ALL.