Biomedical subjects

J M Bogenberger

Publications and source records attributed to J M Bogenberger.

6 recordsLinked to original sources

PubMed2000

Optimization of regulated LTR-mediated expression.

Retroviral vectors are ideally suited to the study of gene function, allowing efficient, stable expression. Many biological systems (e.g., cell cycle, apoptosis) require the use of regulated expression systems. We therefore developed a regulated retroviral vector system, TRA99, based on a tetracycline transactivator-dependent LTR, where the MMLV enhancer was replaced with a tetracycline-response element. Using fluorescence-activated flow cytometric analysis of a destabilized green fluorescent protein to monitor expression levels, we optimized the minimal promoter configuration with respect to both activated and repressed transcription. The TRA99 vectors demonstrate regulated expression with activated levels comparable to those of standard retroviral vectors and repressed levels indistinguishable from background. This was achieved without using an internal promoter cassette, thus retaining the cis-packaging elements requisite for helper-mediated transfer.

Animals↗

PubMed1995

Purification and biologic characterization of plasma-derived megakaryocyte growth and development factor.

The isolation and cloning of the ligand for the cytokine receptor, Mpl, have been recently described. In this report we present details of the purification of this novel cytokine (megakaryocyte growth and development factor [MGDF]) from aplastic canine plasma. Two forms of canine MGDF, with apparent molecular weights of 25 kD and 31 kD and sharing a common N-terminal amino acid sequence, were isolated. The sole contaminant detected in purified 25-kD or 31-kD MGDF was canine Ig. Canine MGDF is characterized as a human megakaryocyte colony-stimulating factor that acts synergistically with human recombinant stem cell factor but not interleukin-3. MGDF also appears to be physiologically regulated in response to platelet demand. In canine and murine models, serum levels of MGDF activity peak during the thrombocytopenic periods after irradiation, 5-fluorouracil, or antiplatelet antisera injections. These data indicate that the megakaryocyte-stimulating activity that accumulates in plasma in response to platelet losses is a novel cytokine that functions through an interaction with the Mpl cytokine receptor.

Amino Acid Sequence↗

PubMed1987

A highly repetitive DNA component common to all Cervidae: its organization and chromosomal distribution during evolution.

In recent work we have isolated and characterized a highly repetitive DNA (MMV satellite IA) from Muntiacus muntjak vaginalis, the species with the most reduced karyotype in the Cervidae family. We have now analysed the genomes of nine related species for the presence of MMV satellite IA components, and have determined their organization and chromosomal distribution. Repetitive satellite IA type DNA is present in all species of the Cervidae, and also in the bovine, but not in a species of the Tragulidae suggesting that these sequences were generated after the phylogenetic separation of Bovidae and Tragulidae. Studies on the organization of the satellite IA DNA in the various species revealed three main repeat lengths: 1400, 1000 and 807 bp. The relative proportion of satellite IA sequences present in any one of the three registers is strikingly different within the various species and can be correlated with the phylogeny of the Cervidae. The chromosomal locations of the satellite IA sequences were determined in seven species by in situ hybridization. It turned out that the chromosomal rearrangements leading to the reduction in the number of chromosomes during karyotype evolution have led to the elimination of satellite I DNA at most locations. In all tandem fusions, the satellite IA sequences located at the centromeres of the ancestral acrocentric chromosomes are lost. In contrast, during the centric fusion that generates the M. m. vaginalis X chromosome satellite IA sequences are amplified. Sequence motifs, which are known to be involved in recombinational events are present in the satellite IA and might have contributed to the unique karyotype variation in the Cervidae.

Animals↗

PubMed1987

The induction of growth factor-independence in murine myelocytes by oncogenes results in monoclonal cell lines and is correlated with cell crisis and karyotypic instability.

Infection of established (IL-3)-dependent hematopoietic cell lines with Abelson murine leukemia virus (A-MLV) abrogates their requirement for IL-3 and leads to non-autocrine growth factor-independent cells. We were interested to determine whether A-MLV can induce IL-3 independence also in non-established cells. To obtain long-term cultures of diploid myelocytes, splenic hematopoietic cells were first infected with MMCV, a murine retrovirus carrying the avian v-myc oncogene. These cultures were superinfected with A-MLV. In three independent experiments, the first growth factor-independent cells appeared between 18 and 43 days after superinfection with A-MLV and represented .02-1% of the population. Furthermore, the cultures that became growth factor-independent were monoclonal for integration of the v-abl gene. These results indicate that the acquisition of growth factor-independence after superinfection of v-myc-expressing cells with A-MLV is a rare event. The low frequency of growth factor-independent cells was not due to a low percentage of infected cells, since 15-25% of the cells were infected with A-MLV after 7 days. The first appearance of growth factor-independent cells coincided with crisis in the cultures, as indicated by a high incidence of cell death and a reduced overall growth rate of the cell populations. These growth factor-independent cells exhibited variable karyotypes, including many that were near-triploid to near-tetraploid. In summary, growth factor-independence induced by super-infection with A-MLV, like that induced by double-infection with v-myc- and v-H-ras-containing viruses, is associated with unstable karyotypes. The growth factor-independent cells show variable ploidy characteristic of cells which survived crisis.

Animals↗

PubMed1986

Autocrine growth and progression of murine X-ray-induced T cell lymphomas.

Primary tumors of X-ray-induced murine T cell lymphomas comprise autocrine, growth factor-dependent cells. We have grown cell lines from primary X-ray-induced thymic lymphomas (PXTLs) under conditions which minimize the progression of the cells from factor dependence to factor independence. All (22) PXTL lines grown secrete a growth factor which supports their own growth and which we will call lymphoma growth factor LGF. LGF-dependent cells are non-tumorigenic or poorly tumorigenic, do not clone in soft agar, have no detectable rearrangements in the c-myc or Pim-1 region and possess near diploid or pseudodiploid karyotypes without evidence for trisomy of chromosomes nos. 15 or 17. PXTL-secreted LGF has no interleukin 1, 2, or 3 activity nor do LGF-secreting cells synthesize detectable IL-1, -2, or -3 mRNA. LGF contains no detectable interferon or GM-CSF activity in specific bioassays. Purified EGF, TGF beta, and interleukin preparations are inactive on LGF-dependent PXTL cells. Thus LGF appears to be a new growth factor that is required for the proliferation of non-progressed T lymphoma cells. Upon progression PXTL cells become growth factor independent, are highly tumorigenic in vivo, clone in soft agar, and assume a near triploid karyotype containing numerous chromosomal aberrations. Thus in X-ray-induced lymphomagenesis an autocrine, LGF-dependent phase precedes the progressed phase characterized by rearrangements in the myc and/or Pim-1 regions as well as by many chromosomal aberrations visible in the karyotype.

Animals↗

PubMed1985

The muntjak satellite IA sequence is composed of 31-base-pair internal repeats that are highly homologous to the 31-base-pair subrepeats of the bovine satellite 1.715.

The nucleotide sequence of a cloned Muntjak satellite IA repeat unit (Muntiacus muntjak vaginalis) was determined. The repeat is 807 base pairs (bp) long. By introducing minor deletions and insertions, the whole sequence of the satellite can be arranged in 27 subrepeats of 31 bp length. Although diverged relative to each other, all subrepeats show a homology of more than 53% with the common consensus sequence. In 29 out of the 31 bp the consensus sequence of the Muntjak satellite subrepeat is identical to the 31-bp subrepeat of the bovine satellite 1.715. This suggests that both satellites are derived from a common ancestral sequence. The results have interesting implications for the evolution of the two satellites.

Animals↗