Cloning and characterization of a novel transcription factor involved in cellular proliferation arrest: PATF.

Cell cycle withdrawal involves several transcription factors such as E2Fs members that play a key role in cell growth control. Here we describe a novel putative bZIP transcription factor isolated from the retina and involved in neuronal proliferation arrest at the terminal differentiation: PATF (Proliferation Arrest Transcription Factor). We show that PATF associates with E2F4 protein and interacts with the E2F consensus site. PATF expression increases with establishment of quiescent state. Furthermore, the nuclear PATF localization like E2F4, depends on cell growth arrest. The decrease of PATF amount, using a retroviral antisense strategy, results in pursued neuroretina cell mitosis. Our results indicate that PATF could be a new molecular signal implicated in the final neuronal cell cycle withdrawal.

CD45 autoantibodies mediate neutralization of activated T cells from lupus patients through anergy or apoptosis.

The objectives were to provide estimates of the prevalence of autoantibody (Ab) directed to CD45 in lupus patients, identify the target autoantigen(s), and determine the ability of such reactivity to mediate neutralization of T lymphocytes. Sera from 64 patients were studied using 2 assays: Western blot and an ELISA with CD45 eluted from 3 cell lines as antigen (U937, Jurkat and Daudi). The role of carbohydrate specificity was investigated using enzyme digestion of blotted glycans, competition with sugars, and inhibition with lectins. Apoptosis was studied through annexin V binding, and cell cycle analysis using the propidium iodide method. AutoAb to CD45 were detected in 16/64 sera (25%) by Western blot, and 21/32 sera (66%) found positive in the ELISA. CD45 purified from Daudi cells was identified in the ELISA, but not in the blot. AutoAb were of the IgM and the IgG isotypes, but not specific for a particular cell type or CD45 isoform: 2 dominant specificities were recognized, one against p180, and another against high MW isoforms. Neuraminidase-induced enhancement of reactivity, together with the inhibitory effect of N-acetyl galactosamine and Dolichos diflorus lectin suggest that the epitopes are carbohydrates. AutoAb which were specific for activated CD4+T cells triggered the annexin V binding, and, in 2 of 4 cases, lymphocytes underwent apoptosis. In conclusion, carbohydrate conformational epitopes may be important as target antigens, and some CD45 autoAb have the capacity to neutralize activated T cells through anergy or apoptosis.

Abnormal distribution of CD45 isoforms expressed by CD4+ and CD8+ T cells in rheumatoid arthritis.

CD45RO+ T cells are referred to as memory or helper-inducer while CD45RA+ T cells are regarded as naive or suppressor-inducer T cells. The former population predominates in the peripheral blood and even more in the synovial fluid of patients with rheumatoid arthritis, to the expense of the latter population. Within the CD45RB+ compartment, there appears to be more of the fully-differentiated than of the early-differentiated CD4+ T cells. In spite of the fact that these lymphocytes are close to undergoing apoptosis, this programmed cell death is inhibited in the rheumatoid synovium.

TSH control of PKA catalytic subunit activity in thyroid cell cultures.

The protein expression and the enzyme activity of the catalytic subunit (C) of the cAMP-dependent protein kinases were studied in porcine thyroid cell primary cultures stimulated with two doses of TSH (0.1 mU/ml and 1 mU/ml) for 1 to 3 days. In TSH-stimulated cells the desensitization of the catalytic subunit activity was accompanied by a simultaneous and parallel decrease of its immunoreactivity. The loss of catalytic subunit was rapid and reached its maximum after 1 day of culture. It is similar in the two subcellular compartments: cytosol and particulate extracts. Contrary to the observed loss of the C subunit protein molecules in TSH-stimulated cells, the expression of the Cbeta subunit mRNA in these cells was increased fivefold compared to controls, while no significant change was observed on the Calpha subunit mRNA. These results suggest that TSH controls the Cbeta subunits of PKA at two levels: at the transcriptional level it increases Cbeta mRNA expression, and at the translational or posttranslational level TSH decreases the amount and the activity of the Cbeta protein molecules.

Autoantibodies to CD45 in systemic lupus erythematosus.

Western blotting with U937 cell extracts as the substrate, and enzyme-linked immunosorbent assays (ELISA) with U937-, Jurkat- and Daudi cell-purified CD45 molecules were used to detect anti-CD45 reactivity in patients with systemic lupus erythematosus (SLE). By immunoblotting, 16 of 64 SLE sera were shown to be positive (25.0%). In the ELISAs, 13 out of 18 SLE sera reacted with the target CD45. Of these, three were not detectable on the blot. Importantly, 12 of these ELISA-positive sera contained IgM and IgG auto-antibodies. Neuraminidase-treatment of U937-precipitated CD45 molecules enhanced the reactivity to most of the isoforms, indicating that the antibodies may bind to asialylated polysaccharides.