The TMC Worldwide Gene Therapy Enrollment Report, end 1996.
Explore the source record for details and available documents.
Clinical Trials as Topic↗
Biomedical subjects
J D Grausz
Publications and source records attributed to J D Grausz.
The TMC Worldwide Gene Therapy Enrollment Report (June 1996).
Explore the source record for details and available documents.
Clinical Trials as Topic↗
Gene mapping of the mammalian genome: the CEPH and Genethon initiative.
Over the past four years, the CEPH (Jean Dausset Foundation) has expanded its linkage mapping effort to include physical mapping and, in 1990, co-founded the Genethon to ensure that a combined physical and genetic map of the entire human genome would be achieved. The Genethon has applied methods developed at CEPH on an industrial scale to accomplish the colossal task of constructing an integrated map. It is the role of such an integrated map to accelerate the search for the genes responsible for inherited diseases, and the results of the past 12 months encourage our optimism that this goal will be realized rapidly. These discoveries are providing not only an approach to the diagnosis of genetically based disease but also some of the first breakthroughs in the area of gene therapy.
Animals↗
Molecular cloning of a 5' segment of the genomic phl gene defines a new breakpoint cluster region (bcr2) in Philadelphia-positive acute leukemias.
Molecular rearrangements of Ph1 chromosome, the hallmark of CML, are clustered in a 5.8-kb DNA segment, the so-called breakpoint cluster region (bcr) of the phl gene that is localized to chromosome 22q11. In Ph1-positive (Ph1+) ALLs, the rearrangements have been shown to involve either the 5.8-kb bcr (called bcr+) or a region upstream of bcr in the 5' end of the phl gene (bcr-). To gain insight into the rearrangements occurring in Ph1+ acute leukemias, a 64-kb DNA fragment from the 5' end of phl was analyzed in order to generate molecular probes covering 40 kb of the phl gene first intron. A panel of seven cases of bcr-Ph1+ acute leukemia (three nonlymphocytic and four lymphocytic) was investigated with these intron 1-derived probes. Strikingly, in six of the seven leukemias, the breakpoints were located in a 10.8-kb DNA segment, defining a new bcr which appears to be specific for Ph1+ acute leukemias. By analogy with the CML bcr region located in the 3' part of the phl gene, we propose to designate this 10.8-kb fragment bcr2.
Chromosome Mapping↗
Sustained IL-2 production by the EL4 subline during continuous phorbol diester stimulation is related to an increase of IL-2-mRNA.
The murine T lymphoma EL4 subline produces large amounts of interleukin 2 (IL-2) upon stimulation with phorbol myristate acetate (PMA). It was found to survive continuous PMA stimulation (10 ng/ml, renewed daily, at 10(6) cells/ml), and to maintain its IL-2 production for several days in a culture medium low in protein. IL-2 production varied from day to day with a peak within the first 2 days of stimulation. PMA stimulation was accompanied by morphologic changes (enlargement and adhesiveness), arrest in cell division and DNA replication, continued RNA synthesis and increased mitochondrial activity. These effects were reversible upon removal of PMA, with a decrease of IL-2 secretion to the background level and a progressive recovery of cell growth. The reverted EL4 cells were again able to produce large amounts of IL-2 upon restimulation with PMA. Growth of EL4 cells was shown not to depend on autologous production of IL-2, and their DNA synthesis could be arrested by aphidicolin without a concomitant accumulation of IL-2 in the supernatant. This indicates that the increase in the amount of free IL-2 is not simply due to reduced consumption by the EL4 cells themselves. PMA appears to act at the level of the expression of the IL-2 gene, since IL-2 mRNA was much more abundant in the IL-2-secreting PMA-treated cells than in untreated controls. Our observations show that considerable amounts of IL-2 can be obtained from a single EL4 cell inoculum and may represent an original model to study the effects of PMA on the differentiation and maturation processes of cells.
Animals↗
Modulation of c-fos and c-myc mRNA levels in normal human lymphocytes by calcium ionophore A23187 and phorbol ester.
A transient expression of the proto-oncogenes c-fos and c-myc is rapidly induced by growth factors or mitogens in different cell types including lectin-stimulated lymphocytes. To elucidate the early events of lymphocyte activation, we determined (by Northern blot analysis) the steady-state levels of c-fos and c-myc mRNA in normal human peripheral blood lymphocytes (PBL) stimulated with the Ca2+ ionophore A23187 and/or 12-O-tetradecanoylphorbol-13-acetate (TPA), whose biological activities are well defined. We report that ionophore A23187 (0.5 microM) or, to a significantly lesser extent, TPA (0.5 ng/ml), neither of which is mitogenic alone at these concentrations, nevertheless can induce a transient accumulation of the proto-oncogene transcripts. These results indicate that a significant accumulation of c-fos and c-myc mRNA can occur without subsequent lymphocyte proliferation. The combination of these two mitogens increases the induced levels of both types of c-onc mRNAs. The level of mRNA accumulation correlates with the degree of proliferation induced by mitogenic combinations of ionophore A23187 and TPA, as measured by [3H]thymidine incorporation. Thus, these compounds act synergistically both with respect to c-fos and c-myc mRNA accumulation and to mitogenicity in human PBL. We also studied the level of c-fos transcripts following stimulation of the T lymphocyte T3/Ti receptor complex, using monoclonal anti-T3 antibodies. We observed that mitogenic doses of anti-T3 also induce an accumulation of c-fos mRNA, whose induction also is synergized by TPA. These results suggest that transient accumulation of c-fos; as well as c-myc, mRNA is a normal event of the immune response. They confirm that Ca2+ ionophore combined with TPA can mimic some molecular features of the early steps of normal human PBL activation.
Antigens, Differentiation, T-Lymphocyte↗
Separation of effector cells mediating antibody-dependent cellular cytotoxicity (ADC) to erythrocyte targets from those mediating ADC to tumor targets.
Murine spleen cells mediate antibody-dependent cellular cytotoxicity (ADC) both to erythrocyte targets in a 51Cr release assay and to syngeneic tumor targets in a microcytotoxicity assay. The effector cells active in the two ADC assays can be separated by passage of the spleen cells through columns of Sephadex G-10 at 37 degrees C. Cells mediating ADC to sarcoma cells did not adhere to the G-10 and were recovered in the column effluent. These nonadherent cells were not cytotoxic to antibody-coated chicken red blood cells. Spleen cells which mediated ADC in a 51Cr release assay to the red cell targets adhered to G-10. Adherent effector cells could subsequently be recovered from the columns by elution with 5 X 10(-4) M EDTA.
Animals↗