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C J Larsen

Publications and source records attributed to C J Larsen.

At least 73 records · Page 4Linked to original sources

A new gene, BCM, on chromosome 16 is fused to the interleukin 2 gene by a t(4;16)(q26;p13) translocation in a malignant T cell lymphoma.

A t(4;16)(q26;p13.1) chromosome translocation found in tumour cells from a patient with a T cell lymphoma was shown to rearrange the interleukin 2 gene, normally located on chromosome band 4q26, with sequences from chromosome band 16p13.1. A cDNA library of tumour cells was screened with an interleukin 2 gene-specific probe. Three clones were isolated, which consisted, from 5' to 3', of the three first exons of the interleukin 2 gene followed by a 16p13 in-frame sequence encoding 181 amino acids. A probe derived from this sequence detected a 1.2 kb transcript in various cell lines exhibiting mature B lymphoid cell features, but this was not detected in other cell lines representative of other haematopoietic lineages, or in other organs. For this reason, the novel gene was termed BCM for B cell maturation. The open reading frame of BCM normal cDNA predicted a 184 amino acid protein with a single transmembrane domain which had no homology with any protein sequence stored in data banks. Our data indicate that BCM is a new gene whose expression coincides with B cell terminal maturation.

Amino Acid Sequence↗

Molecular rearrangements of the MYL gene in acute promyelocytic leukemia (APL, M3) define a breakpoint cluster region as well as some molecular variants.

Genomic DNA probes generated from the retinoic acid receptor alpha (RARA) gene located on chromosome 17 and from the MYL gene located on chromosome 15 were used to study the chromosome 15 breakpoints resulting from the t(15; 17) translocation in 26 patients with acute promyelocytic leukemia (APL). In 20 out of 22 patients with a detectable MYL rearrangement, the breakpoints were clustered within a 4.4 kb segment designated MYLbcr. The two remaining patients exhibited a more 5' rearrangement at about 10 kb upstream of the MYLbcr region, implying the lack of at least one MYL gene exon in the resulting MYL-RARA fusion gene. The variation of chromosome breakpoints within the MYL gene may explain size heterogeneity previously observed in some MYL-RARA fusion transcripts expressed in APL cells.

Blotting, Southern↗

Hemizygous deletion of the 3' end of the RB1 gene in a case of Philadelphia positive-acute lymphoblastic leukemia.

In order to address a possible role for the human retinoblastoma susceptibility (RB1) gene in hematopoietic malignancies, 34 cases of different types of leukemia without chromosomal abnormalities at band 13q14 were analyzed by Southern blot hybridization. A large deletion encompassing exon 18-27 was detected in one case of Ph1 positive-acute lymphoblastic leukemia. Interestingly, this deletion appeared to be similar to another deletion recently reported in a leukemic cell line derived from a T-ALL. No rearrangement was detected in all other leukemias.

Adult↗

t(18;22)(q21;q11) with rearrangement of BCL2 as a possible secondary change in a lymphocytic lymphoma.

We report a lymphocytic lymphoma showing a combination of two characteristic neoplasia-associated chromosomal changes: trisomy 12, commonly observed in chronic lymphocytic leukemia and lymphocytic lymphoma, and t(18;22)(q21;q11), a variant form of the t(14;18)(q32;q21) found in most follicular lymphomas. Southern blot analysis was performed using probes for the 5' end of the BCL2 gene (18q21) and for the J lambda as well as C lambda immunoglobulin genes (22q11). With these two probes, a unique rearranged fragment was detected. Thus the t(18;22)(q21;q11) can be considered as a variant translocation of t(14;18)(q32;q21). The karyotypic analysis supports the assumption that in our case trisomy 12 occurred first, and t(18;22) appeared during tumor progression as part of the clonal evolution. This is at variance with the typical t(14;18), which has never been found to occur as a secondary change.

Blotting, Southern↗

A novel 43-kDa glycoprotein is detected in the nucleus of mammalian cells by autoantibodies from dogs with autoimmune disorders.

We have characterized a new antibody specificity in a panel of sera from dogs developing systemic lupus erythematosus (SLE) or clinically related autoimmune disorders. This antibody stains in a speckled fashion the nucleus of cells of different mammalian origins. The target antigen is a basic (pI 9.2) nuclear polypeptide with an apparent molecular weight of 43 kDa (p43) which is detected in various mammalian cell nuclei. p43, as studied in HeLa cells, appears to be cell cycle-independent. It is released from nuclei by salts (0.5 M NaCl or 0.25 M ammonium sulfate). Upon subfractionation of nuclear components, p43 is found in the fraction containing HnRNPs and is recovered in immunoprecipitates obtained with 4F4 monoclonal antibody to HnRNP C proteins. Immunoelectron microscopy revealed that p43 is concentrated over the dense chromatin periphery and interchromatin granule clusters. Another important feature of p43 is its ability to specifically bind wheat germ agglutinin lectin but not concanavalin A nor Ulex europaeus I, supporting the notion that p43 is a glycoprotein bearing an N-acetyl-glucosamine moiety. Consistent with this result, a radio-active p43 band is specifically immunoprecipitated by canine anti-p43 autoantibodies from HeLa cells metabolically labeled with [14C]glucosamine. Finally, anti-p43 antibodies do not immunoprecipitate SnRNA, indicating that p43 has no apparent association with SnRNPs.

Animals↗

Studies of BCR rearrangements in Philadelphia-positive acute leukemia.

Five patients with Philadelphia-positive acute leukemia were cytogenetically and molecularly investigated in order to determine the localization of the breakpoints on chromosome 22. Rearrangements of the bcr segment were detected in one case with acute mixed leukemia in a child. Rearrangements in the BCR gene first intron, the so-called bcr2 and bcr3 regions, were detected in two other cases, one with an acute lymphoblastic leukemia (ALL) and one with mixed acute leukemia. No molecular rearrangement could be detected in the last two cases, an ALL and a T-cell acute lymphoblastic leukemia with a t(2;22) translocation.

Acute Disease↗

c-myc gene expression in a leukemic T-cell line bearing a t(8;14) (q24;q11) translocation.

We have examined the expression of the c-myc protooncogene in human T-cell leukemic KE-37R cells carrying a t(8;14) (q24;q11) translocation. The breakpoint on chromosome 8 is located at 2.2 kb downstream of c-myc exon 3 and the 3' part of the TcR-alpha gene (14q11) has been juxtaposed to c-myc. Our results showed that the steady-state levels of c-myc RNA transcripts were increased and the P1/P2 ratio of c-myc promoter utilization did not change, indicating that preferential utilization of P2 was maintained in the rearranged gene. High levels of electrophoretically normal p64 and 67 c-myc proteins were detected and both products kept their instability. In addition, transcription from promoter P0 was not detectable. Our results suggest that the activation of the gene is likely to result from its juxtaposition to the enhancer element of the TcR-alpha gene located downstream of the Ca region which stimulates constitutive synthesis of normal c-myc transcripts from the rearranged allele.

Blotting, Northern↗

Monoallelic deletions of the P53 gene in Chinese patients with chronic myelogenous leukemia in blastic crisis.

The configuration of the P53 tumor suppressor gene was investigated in 43 Chinese patients with chronic myelogenous leukemia (CML), 32 in chronic phase and 11 in blastic crisis. No obvious rearranged DNA band was detected in Southern blot patterns from patients at both stages of the disease. However, P53 gene deletion events were observed in 4 out of 11 cases in blast crisis. This finding was associated with a cytogenetically identifiable chromosome 17p deletion, iso(17q), in only one out of 4 cases.

Adult↗

The retinoic acid alpha receptor gene is frequently disrupted in its 5' part in Chinese patients with acute promyelocytic leukemia.

The t(15;17)(q22;q11-q21) translocation, a hallmark for acute promyelocytic leukemia (APL), has recently been shown to disrupt the retinoic acid-alpha receptor (RARA) gene localized on band 17q21. Leukemic cell DNAs from 16 Chinese patients with APL were analysed by Southern blot hybridization with probes covering the 5' and the 3' parts of the gene. In ten patients, the breakpoints were concentrated within a 5.5 kb EcoRI fragment containing part of intron 1, whereas no rearrangement was detected in the six remaining patients. Genomic amplification of the RARA gene was observed in one case. When RNA was available (six patients), abnormal-sized RARA gene transcripts were observed. Interestingly, no rearranged transcript was found in cells from a patient during a 6-day treatment with all-trans retinoic acid, while bone marrow cells still possessed the translocation.

Adolescent↗

Two site-specific deletions and t(1;14) translocation restricted to human T-cell acute leukemias disrupt the 5' part of the tal-1 gene.

Analysis of several cases of t(1:14)(p32;q11) translocation present in 3% of T-cell acute leukemias (T-ALL) has revealed the tal-1 gene. This gene encodes a helix-loop-helix protein. It has been found to be expressed in normal bone marrow and in leukemic T-cell and erythroleukemia cell lines, but not in normal T cells. Recently, a site-specific deletion, tald, renamed tald1 in this paper, has been detected in a high proportion of pediatric T-ALL, which arose by a site-specific DNA recombination between tal-1 and a new locus termed SIL. In this study we searched for structural rearrangements within tal-1 in a panel of 134 non-selected leukemic patients (including 66 with T-ALL). Only 6% of patients with T-ALL harbored the tald1 deletion. A second specific deletion termed tald2 was observed in another 6% of T-ALL patients; it involves another site within tal-1 plus the same site as tald1 in the SIL locus. Similarly to tald1 deletion, tald2 junctions harbor structural characteristics that are reminiscent of aberrant recombinase activity. Moreover, we report a detailed analysis of the tal-1 gene structure. Transcription analysis and in vitro translation data are consistent with the differential expression of several TAL-1 protein species containing the HLH motif but differing in their amino terminus. Taken together, our data indicate that t(1;14) translocations and both tald deletions disrupt the 5' part of the tal-1 gene, placing its entire coding sequences under the control of the regulatory elements of the TCR-delta gene or the SIL gene, both of which are normally expressed in T-cell lineage.

Amino Acid Sequence↗

A variant translocation t(2;18) in follicular lymphoma involves the 5' end of bcl-2 and Ig kappa light chain gene.

We have examined three cases of human lymphoma bearing a t(2;18)(p11;q21) chromosome translocation. The bcl-2 gene appeared to be rearranged in all three cases and breakpoints were clustered in the 5' flanking region of the gene. In all three cases, bcl-2 was juxtaposed to J segments of the Ig kappa gene. This juxtaposition of the bcl-2 and Ig kappa genes is very similar to the variant chromosome translocations of Burkitt lymphoma that juxtapose the c-myc locus to IgL genes.

Adult↗

Inactivation of the p53 gene expression by a splice donor site mutation in a human T-cell leukemia cell line.

An abnormally sized 3.5 kb p53 transcript was detected in the KE-37R human leukemic T-cell line in which no p53 protein could be detected by immunoprecipitation. S1 nuclease protection experiments and sequencing analysis indicated conservation of the entire intron 4 (755 bp) in the 3.5 kb transcript and the presence of a G to A substitution in the last exonic nucleotide of the splice donor site. These data support the notion that p53 gene inactivation by point mutations in splice junctions also exists in hemopoietic neoplasia.

Animals↗

Structural organization of the hCTLA-1 gene encoding human granzyme B.

Cytotoxic T lymphocytes (CTLs) and natural killer/lymphokine-activated cells produce granzymes, a family of serine esterase proteins located in cytoplasmic granules. These might be involved in different cytotoxic pathways. We report the structural organization of the human gene encoding granzyme B (hCTLA-1). A 4.75-kb genomic DNA fragment containing all the sequences of granzyme B-encoding cDNA clones has been sequenced. The gene is composed of five exons and four introns. A comparison with the genomic organization of murine CCP1/CTLA-1 showed very similar structure and a 76% nucleotide homology in the coding sequences. This suggests that both genes may have a common ancestor. No typical regulatory element was detected in the 1160 bp upstream from the ATG start codon. The detection of a second locus related to hCTLA-1 is also described.

Amino Acid Sequence↗

Two distinct mechanisms for the SCL gene activation in the t(1;14) translocation of T-cell leukemias.

Molecular study of a t(1;14)(p32;q11) translocation found in an acute T-cell leukemia (Kd cells) with a relatively mature phenotype is reported. Complex DNA rearrangements were characterized in the TCR alpha/delta locus. Besides a productive V alpha/J alpha assembly found on the normal allele, two deletions within the J alpha cluster were identified in the translocated allele. The translocation breakpoints involved the TCR delta gene on chromosome 14 and the SCL locus on chromosome band Ip32 that was recently shown to be activated by the t(1;14) translocation of the DU 528 leukemic cell line. Significantly, both Kd and DU 528 translocation breakpoints were located at the boundaries of D delta or J delta segments and were clustered in a 10 kb genomic fragment of the SCL gene. The presence of recombination signal motifs (heptamer-12/23 bp spacer-nonamer) on both normal chromosome partners, and N nucleotide addition on both derivative chromosomes involved the recombinase system in the translocation event. The SCL locus was highly expressed as a 5 kb transcript in Kd cells and, as already reported, as a 2 kb transcript in DU 528 cells. Importantly, a 5 kb SCL transcript was also detected in immature nonlymphoid hematopoietic cells but not in normal mature T cells, suggesting that it might correspond to the normal SCL transcript. Taken together, our data support the notion that the involvement of the SCL gene in the leukemogenic process may occur through overexpression of an apparently normal transcript (Kd cells) or expression of a truncated RNA (DU 528 cells).

Amino Acid Sequence↗

Loss of heterozygosity of the L-myc oncogene in human breast tumors.

Recent studies suggest that loss of heterozygosity may play an important role in various human neoplasia. Cytogenetic abnormalities detected in primary breast tumors led us to examine breast tumor DNAs for deletions. In the present study, we demonstrate, using restriction fragment length polymorphism (RFLP) analysis at the L-myc proto-oncogene (chromosome 1p32), a frequent loss of heterozygosity in primary breast tumor DNAs (55 out of 152 informative tumor DNAs). Most of these deletions appear to be limited to chromosome 1p. No correlation was observed between this genetic alteration and several parameters of each patient's history or characteristics of the tumor. However, a significantly (P = 0.011) shorter survival period after relapse was observed for patients with loss of heterozygosity at L-myc in primary tumor DNAs compared with patients with tumor DNAs lacking this alteration.

Blotting, Southern↗

Isolation of a Friend recombinant polytropic virus with a T-cell-restricted leukemogenicity.

Hematopoietic malignant cells of various types were isolated from ICFW mice inoculated as newborn or adult with F-MuLV and grafted into recipient mice. After repeated in vivo cell transplants, several recombinant polytropic viruses (also termed MCF or dualtropic viruses) were isolated from tumors by limiting dilution. Two virus isolates designated RA1-17 and EA1-17 were recovered from the spleen and the omentum, respectively, of the same grafted animal. When inoculated into newborn mice, RA1-17 induced erythroblastosis similar to that induced by other Friend recombinant polytropic viruses. Remarkably, EA1-17 induced T-cell leukemia after a short latency. To our knowledge, this is the first description of a polytropic recombinant virus with a T-cell tropism isolated after inoculation of F-MuLV.

Animals↗

[Serum antibodies in dogs with autoimmune disorders recognize 40 kd glycoprotein in the nucleus of mammalian cells].

We report a new antibody specificity in 15 sera recovered from a group of dogs developing systemic lupus erythematosus (SLE) or clinically related disorders. This antibody stains in a speckled fashion the nucleus of human Hep-2 cells. Immunodiffusion tests with saline extracts of rabbit thymus showed that all 15 sera generate a common precipitation line which crosses the lines from reference sera to Sm, SS-A/ro, SS-B/La, and RNP antigens. The target nuclear antigen is a 40 kD polypeptide (p40). An important property of p40 resides in its ability to bind specifically Wheat Germ Agglutinin lectin but not Concanavalin A, supporting the notion that the antigen is a glycoprotein bearing a N-acetylglucosamine moiety.

Animals↗