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J D Norton

Publications and source records attributed to J D Norton.

At least 73 records · Page 4Linked to original sources
PubMed1990

Immunoglobulin heavy chain variable region family usage is independent of tumor cell phenotype in human B lineage leukemias.

During B cell development, immunoglobulin heavy chain (IgH) variable region (VH) genes are rearranged and expressed in a programmed manner and accumulating evidence suggests recurrent utilization of developmentally restricted VH genes in malignant B lymphoid populations. We have used polymerase chain reaction gene amplification in conjunction with a panel of VH family-specific amplimers to directly compare the repertoire of VH region rearrangement in mature, CD5+ B cell chronic lymphocytic leukemia with that in immature, CD5 B lineage acute lymphoblastic leukemia. The results revealed a diverse pattern of VH family utilization common to both disease groups in which VH regions most proximal to the IgH joining locus were preferentially rearranged relative to their family sizes with recurrent utilization of several known developmentally restricted VH genes in close to germ-line configuration. These results indicate that biased VH family usage is independent of tumor cell phenotype in B lineage leukemias. This bias may reflect similar stages or compartments in normal B lymphopoiesis from which diverse types of B cell malignancy may arise. Moreover, since blast cells in acute lymphoblastic leukaemia do not express functional immunoglobulin, we infer that the tumor cell-associated VH family repertoire is determined through antigen-independent mechanisms.

Antigens, Differentiation↗
PubMed1990

Detection of immunoglobulin gene rearrangement in B lymphoid malignancies by polymerase chain reaction gene amplification.

Immunoglobulin heavy chain gene rearrangement serves as a marker of cell lineage and clonality in B lymphoproliferative disorders. We have used polymerase chain reaction (PCR) gene amplification to detect immunoglobulin gene rearrangements involving VH251, a heavy chain variable region preferentially utilized in B lymphoproliferative disorders. Using synthetic amplimers derived from VH251 and the heavy chain joining region, under conditions of high stringency, a homogeneous VH251-specific fragment of approximately 350 bp could be amplified from leukaemic DNA. Of 53 cases of B lineage acute lymphoblastic leukaemia screened for VH251 rearrangement by PCR, 10 were positive. A background level of VH251 rearrangement could also be amplified from normal peripheral blood and bone marrow DNA, but a VH251 rearranged leukaemic clone representing 0.01% of bone marrow mononuclear cells could be readily detected. The application of PCR to detect immunoglobulin gene rearrangement involving VH251 and potentially other preferentially utilized V regions provides a sensitive method both for tracking malignant B cells and for the study of normal B cell developmental pathways through which B lineage malignancies arise.

Blotting, Southern↗
PubMed1990

The t(9;22) translocation in Philadelphia-positive essential thrombocythaemia does not involve the T lymphocyte lineage.

A 42-year-old woman presented with clinical and haematological features of essential thrombocythaemia (ET). Cytogenetic investigation revealed a standard t(9;22) Philadelphia translocation in all evaluable metaphases which persisted throughout treatment. Gene probe analysis of the chromosome 22 breakpoint cluster region (bcr) locus revealed a breakpoint mapping between exons 1 and 3 of the bcr gene, consistent with a standard bcr-abl translocation as found in chronic myeloid leukaemia (CML). Moreover, in separate cell populations, the bcr breakpoint was demonstrable in DNA from granulocytes but not in T cells from either peripheral blood or bone marrow. Since ET is known to be a clonal disorder with a multipotential stem cell origin, these findings suggest that, as in CML, the bcr-abl hybrid gene arises through translocation in a multilineage stem cell which does not involve the T lymphocyte lineage.

Adult↗
PubMed1990

Patterns of nuclear proto-oncogene expression during induced differentiation and proliferation of human B chronic lymphocytic leukaemia cells.

Phorbol ester-induced differentiation of human B-chronic lymphocytic leukaemic cells was found to be preceded by a rapid transient induction in expression of the c-jun proto-oncogene, which paralleled that of c-fos. Induced expression of c-myc but not of c-fos/c-jun proto-oncogenes was markedly higher in a proliferating variant leukaemic cell population compared with that seen in typical lymphocytic leukaemia cells. These data suggest that the c-fos/c-jun nuclear oncogenes play a role in induced differentiation, whilst c-myc is more important in the proliferative response of B lymphocytes.

B-Lymphocytes↗
PubMed1989

Abnormalities in T-cell associated rearrangement of T-cell receptor gamma chain genes in B-cell chronic lymphocytic leukemia.

Non-random rearrangement of genes encoding the gamma chain of the T-cell antigen receptor is known to occur in a developmentally regulated fashion during thymocyte differentiation. We show here that peripheral blood T lymphocytes from patients with B-cell chronic lymphocytic leukemia display marked abnormalities in the spectrum of gamma variable region gene rearrangements compared with normal peripheral blood. Of seven patients studied, all displayed a reduction of rearrangements into V9 (subgroup V II), while two cases had, in addition, over-representation of specific rearrangements involving subgroup V I; one V 2/4, the other V 5. These observations have a number of implications relevant to mechanisms of disease pathogenesis, and may provide new insight into the basis of T-cell dysfunction that is frequently associated with this condition.

Blotting, Southern↗
PubMed1989

T-cell receptor gene rearrangement in B-cell non-Hodgkin's lymphoma: correlation with methylation and expression.

Two of 19 B-cell non-Hodgkin's lymphoma (NHL) were found to have clonally rearranged T-cell receptor (TCR) beta chain genes with a germline TCR-gamma gene configuration. The inappropriate rearrangements had a similar structure to the TCR-beta rearrangements described in B-cell chronic lymphocytic leukemia. All cases analysed displayed a consistent germline transcription of TCR-beta and alpha chain genes which was independent of rearrangement. Consistent with this, all cases showed some degree of hypomethylation of the TCR-beta chain locus similar though distinguishable from the pattern seen in mature T cells, but quite different from that seen in normal mature B cells. Taken together, these data suggest that most NHL's arise from an immature B-lymphocyte precursor at a stage of differentiation where lineage commitment is incomplete.

Antigens, Differentiation↗
PubMed1989

Chromosome 22 breakpoints in variant Philadelphia translocations and Philadelphia-negative chronic myeloid leukemia.

The standard t(9;22)(q34;q11) found in Philadelphia (Ph) chromosome positive chronic myeloid leukemia (CML) involves a highly restricted (5.8 kb) chromosome 22 breakpoint cluster region (bcr), which results in the formation of a chimeric gene comprising exons from the 5' end of bcr and protooncogene c-abl coding sequences from chromosome 9. In a survey of 21 patients with hematologic and clinical features of CML we detected rearrangement of the chromosome 22 bcr by gene probe analysis in all cases, including 16 with a standard t(9;22), two with variant Ph translocations [t(10;22)(q26;q11);t(11;22)(p15;q11)], one with a complex Ph translocation [t(9;11;22)(q34;q13;q11)], one with a complex translocation and a masked Ph[t(9;14;22) (q34;q24;q11)], and one Ph-negative case with a t(1;9)(p32;q34). These observations further substantiate the suggestion that, despite karyotypic heterogeneity, a common underlying molecular lesion, the bcr-abl gene chimera, is involved in the disease pathogenesis of CML.

Adult↗
PubMed1989

Rearrangement of TCR gamma chain gene involving JP1 suggests early thymocyte origin of peripheral T-cell lymphoma.

Peripheral T-cell lymphomas (PTL) are morphologically and immunophenotypically heterogeneous. We have examined a series of cases to determine whether this heterogeneity is reflected at the level of developmentally specific T-cell receptor (TCR) gene rearrangement. 4 of 5 cases had clonal rearrangements of TCR beta and gamma chain genes; one of these also had a probable DQ52-J immunoglobulin heavy chain gene rearrangement. 2 of the 4 TCR gamma gene rearrangements involved the most 5' J region, JP1, a characteristic of immature thymocytes. These 2 cases also had immunophenotypic features of immaturity. Taken together, our results suggest that TCR gene rearrangement is correlated with surface marker data and shows that in some cases PTL may arise from a very early stage of thymocyte maturation.

Antigens, Surface↗
PubMed1989

Clonal evolution in Ph-negative, bcr-positive chronic myeloid leukaemia before and after bone marrow transplantation.

A 26-year-old man, who presented with bilateral fundal haemorrhages, was found to have chronic myeloid leukaemia (CML). The Ph chromosome was not present but a clone with t(1;9) (p32;q34) was detected. On referral for bone marrow transplant (BMT) he was found to be in accelerated phase with clonal evolution in three cell lines inv(3)(q21q26); inv(3)(q),i(17q); inv(3q)+8. Molecular investigation revealed a breakpoint on chromosome 22 within the breakpoint cluster region (bcr) similar to that found in Ph+ cases. After BMT, from an HLA-identical sister, successful engraftment (46,XX) was accompanied by evidence of a residual host cell population with further evolution (del(7)(q22)) and persistence of the bcr+ clone. Acute myeloid leukaemia, detected 5 months later, was associated with predominance of the clone 46XY,t(1;9),inv(3q),del(7)(q22) which failed to respond to treatment and the patient died 6.5 months after BMT. This case indicates that BMT, after the acquisition of additional chromosomal change in accelerated phase, may fail owing to persistence of the leukaemic clone. In addition the BMT conditioning regimen may produce further abnormalities which confer drug resistance on the persisting clone, which can emerge as an intractable myeloid blast crisis.

Adult↗
PubMed1989

Interleukin 2 induced non MHC-restricted killing of herpes simplex type-1 (HSV-1) infected allogeneic and autologous lymphoblasts.

Peripheral blood lymphocytes (PBL) from herpes simplex type-1 (HSV-1) seropositive (sero+) and seronegative (sero-) individuals were assessed before and after in vitro culture with IL-2 for their ability to kill different tumor cells and to kill HSV-1 infected PHA lymphoblasts using a 4 hr 51Cr release assay. Fresh PBL from either sero+ or sero- individuals had little cytotoxic activity against HSV-1 infected autologous PHA lymphoblasts or against different NK-insensitive tumor targets. When PBL from either sero+ or sero- individuals were incubated 3-4 days in vitro with IL-2, significant levels of cytotoxic activity was observed against both allogeneic and autologous HSV-1 infected targets and against different NK-insensitive tumor cells. When comparing PBL from sero+ versus sero- individuals for IL-2 induced cytotoxic activity against HSV-1 infected autologous targets, PBL from sero+ individuals always had significantly higher levels of cytotoxic activity. Allogeneic HSV-1 infected targets were also killed by IL-2 activated PBL obtained from either sero+ or sero- individuals, but IL-2 activated PBL from sero+ individuals showed higher cytotoxic activity on HSV-1 infected autologous targets when compared to HSV-1 infected allogeneic targets. When IL-2 induced cytotoxic effector cells were treated with HSV-1 for 1 hr and recultured with IL-2, significant inhibition of cytotoxic activity against all target cells was observed. Greater than 50% inhibition of cytotoxic activity was observed when IL-2 induced cytotoxic effector cells were treated with HSV-1 and cultured for 6 hr.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Activation of Harvey ras oncogene by mutation at codon 12 is very rare in hemopoietic malignancies.

Point mutations within codon 12 of the Harvey (H-) ras proto-oncogene have recently been implicated in the progression of hemopoietic malignancy, particularly chronic myeloid leukemia. We have analyzed DNA from 170 cases of acute and chronic leukemia by using a restriction fragment length polymorphism. No evidence for clonal allelic H-ras codon 12 activation was found among these cases, which included 23 cases of chronic myeloid leukemia, 12 of which were in accelerated phase or blastic transformation. These data suggest that H-ras codon 12 mutations occur infrequently in hemopoietic neoplasms generally and may be less important in disease progression than has been previously suggested.

Base Composition↗
PubMed1989

Analysis of ras gene mutations and methylation state in human leukemias.

We have screened a large series of primary human leukemias for activating point mutations at codons 12, 13 and 61 of the N-ras and K-ras proto-oncogenes and at codons 12 and 61 of the H-ras proto-oncogene by using panels of oligonucleotide probes in conjunction with polymerase chain reaction gene amplification. 13 of 64 (20%) acute lymphoblastic leukemia cases had ras gene mutations mostly involving N-ras codon 12/13, G-A (gly-asp) transitions. Consistent with previous studies, a comparable pattern and frequency of ras mutation was found amongst 45 cases of acute myeloid leukemia and myelodysplasia. By contrast, of 30 cases of mature B cell chronic lymphocytic leukemia, only one in terminal prolymphocytoid transformation harboured an activated ras gene. These patterns of mutation did not correlate with ras gene methylation state, a finding not obviously compatible with differential gene accessibility being an important determinant of ras gene mutation patterns in leukemogenesis. Our data suggest that activated ras is more important in tumourigenesis of immature than mature lymphocyte progenitors whilst similar mechanisms associated with aetiology and/or target cell susceptibility probably underlie the similar patterns of ras gene mutations seen in acute leukemias of both myeloid and lymphoid cell lineages.

Blotting, Southern↗
PubMed1988

The genomic DNA organisation and evolution of a retrovirus-transmissible family of mouse (VL30) genetic elements.

The sequence organisation of endogenous VL30 elements in the mouse genome was investigated by using a cloned representative of a retrovirus-transmissible VL30 cDNA. The majority of dispersed VL30 sequences could be assigned to a proviral-like structure 5.2-5.3 kbp long and bounded by long terminal repeats (LTRs). The existence of a hierarchy of evolutionarily conserved elements was rather limited and sequence heterogeneity between different elements was randomly distributed. However, the retrovirus-transmissible class of VL30 element was found to represent a distinct minority subgroup distinguishable by restriction sites and size (4.6-4.9 kbp long). Analysis of sequence conservation showed that VL30 elements display a more rapid turnover than endogenous murine leukaemia virus-related proviral sequences, and that VL30 LTRs show the most limited evolutionary distribution. Although discrete subsets of VL30 unique sequence were conserved in different rodents, the location of conserved regions was found to be variable, arguing against the presence of a functionally conserved protein coding region. These observations support the hypothesis that high frequency recombination, probably occurring during reverse transcription and the accompanying processes of duplicative transposition and amplification, have been a major determinant in the mode of evolution of the VL30 gene family.

Animals↗
PubMed1988

Temporal and tissue-specific expression of distinct retrovirus-like (VL30) elements during mouse development.

We have analyzed the expression of VL30 retroviral RNA transcripts during mouse embryogenesis. VL30 RNA was found in all tissues examined from mid-gestation, but increased dramatically at later times in the extraembryonic amnion and visceral yolk sac, and to a lesser extent in embryonic liver. The overall temporal and tissue-specific pattern of expression was unlike that of other retrotransposon gene families. S1 nuclease mapping experiments which distinguish transcripts from distinct VL30 elements suggest that they are independently regulated in different tissues during mouse development.

Animals↗
PubMed1988

Use of the X-chromosome linked hypoxanthine phosphoribosyl transferase gene as a marker of cell monoclonality in hemopoietic malignancies.

The X-chromosome linked gene encoding hypoxanthine phosphoribosyl transferase (HPRT) has been reported to provide a novel approach to the investigation of cell monoclonality in females based on non-random methylation-sensitive restriction enzyme cleavage of the active vs inactive HPRT alleles and a polymorphic Bam HI restriction site to distinguish the maternal and paternal gene copies. In a survey of 80 females, which included 65 cases of hemopoietic malignancy and 15 normal individuals, we found only nine (11.3%) to be heterozygous (informative) for the Bam HI polymorphism. Monoclonality was demonstrable by HPRT gene analysis in five of six informative cases of leukemia, the exception being a case of acute myeloid leukemia which displayed anomalous methylation of the HPRT gene. Our studies suggest that the applicability of the HPRT gene probe strategy may be limited by (1) the low frequency of informative cases and (2) potential inappropriate methylation of the HPRT gene in a proportion of cases.

Clone Cells↗
PubMed1988

Mixed acute leukaemias.

In recent years immunophenotyping and analysis of clonal rearrangement of immunoglobulin and T-cell antigen receptor genes have proved valuable for the diagnosis and classification of leukaemia. These techniques aid in the assignment of cell lineage in cases of acute leukaemia in which the standard FAB criteria of morphology and cytochemistry do not reveal clear lymphoid or myeloid phenotype. These new techniques have also revealed that the leukaemic blasts in a sizable minority of otherwise typical cases of acute leukaemia express 'inappropriate' lineage-associated markers and have been termed mixed acute leukaemias. The spectrum of characteristics encompassed by mixed acute leukaemias ranges from fairly common cases expressing one or two inappropriate markers to the more extreme, rare cases of acute leukaemia termed 'hybrid' in which a truly scrambled picture is seen. A subgroup of these mixed cases have two distinct populations of blasts, e.g. one lymphoid and the other myeloid. These observations raise a number of issues about the cell of origin of these leukaemias and about the mechanisms controlling the developmental regulation of expression of different lineage-associated markers. In addition, accumulating evidence suggests that inappropriate expression of markers may identify sub-groups of both acute myeloid and lymphoblastic leukaemia with an inferior prognosis.

Diagnosis, Differential↗
PubMed1988

Absence of Kirsten-ras oncogene activation in B-cell chronic lymphocytic leukemia.

By using a combination oligonucleotide probe hybridization and restriction enzyme polymorphism analysis, a series of 48 cases of B-cell chronic lymphocytic leukemia were investigated for activating point mutations at codons 12, 13 and 61 of the K-ras proto-oncogene. A small series of acute leukemias (seven with acute lymphoblastic leukemia (ALL), 11 with acute myeloid leukemia (AML)) were examined in parallel. None of the cases of B-CLL contained detectable activating mutations of the K-ras gene at codon 12 (GGT-gly----GCT-ala) was detected at presentation. In both cases of acute leukemia, the mutation was restricted to one allele and could not be detected in remission samples. Those data suggest that activation of members of the ras oncogene family, typified by K-ras, may be less important in disease pathogenesis in leukemias such as B-CLL that arise from a more committed progenitor.

Aged↗