Circulating apoptotic microparticles in systemic lupus erythematosus patients drive the activation of dendritic cell subsets and prime neutrophils for NETosis

\u3cp\u3eOBJECTIVE: Circulating chromatin-containing apoptotic material and/or neutrophil extracellular traps (NETs) have been proposed to be an important driving force for the antichromatin autoimmune response in patients with systemic lupus erythematosus (SLE). The aim of this study was to determine the exact nature of microparticles in the circulation of SLE patients and to assess the effects of the microparticles on the immune system.\u3c/p\u3e\u3cp\u3eMETHODS: We analyzed microparticles isolated from the plasma of patients with SLE, rheumatoid arthritis (RA), and systemic sclerosis (SSc), as well as from healthy subjects. The effects of the microparticles on blood-derived dendritic cells (DCs) and neutrophils were assessed by flow cytometry, enzyme-linked immunosorbent assay, and immunofluorescence microscopy.\u3c/p\u3e\u3cp\u3eRESULTS: In SLE patients, we identified microparticles that were highly positive for annexin V and apoptosis-modified chromatin that were not present in healthy subjects or in RA or SSc patients. These microparticles were mostly CD31+/CD45- (endothelial), partly CD45+/CD66b+ (granulocyte), and negative for B and T cell markers. Microparticles isolated from the plasma of SLE patients increased the expression of the costimulatory surface molecules CD40, CD80, CD83, and CD86 and the production of proinflammatory cytokines interleukin-6, tumor necrosis factor, and interferon-α by blood-derived plasmacytoid DCs (PDCs) and myeloid DCs (MDCs). SLE microparticles also primed blood-derived neutrophils for NETosis. Microparticles from healthy subjects and from RA or SSc patients exhibited no significant effects on MDCs, PDCs, and NETosis.\u3c/p\u3e\u3cp\u3eCONCLUSION: Circulating microparticles in SLE patients include a population of apoptotic cell-derived microparticles that has proinflammatory effects on PDCs and MDCs and enhances NETosis. These results underline the important role of apoptotic microparticles in driving the autoimmune response in SLE patients.\u3c/p\u3

An active mechanism flanks and modulates the export of the small ribosomal subunits.

Item does not contain fulltextThe modalities of export of the ribosomal subunits from the nucleolus to the nuclear pores have been only partially clarified since it is not yet clear whether the movements depend purely on diffusion or also from an active process. Recently, we suggested the existence of an active transport mechanism of a subset (10-12%) of the small ribosomal subunits (SSU) (Cisterna et al. in 2006, Faseb J). Here, we give further evidence that an active, motor protein-mediated process exists for the SSU transport from the nucleolus to the nuclear pore. We demonstrate that the blockade of ATP synthesis and antibody-mediated inhibition of nuclear myosin or actin induce structural and functional modifications of the nucleolus, suggestive of transcriptional activity decrease. Moreover, both treatments induce a significant retention of RNA inside the nucleus and an accumulation of ribosomal subunits in the granular component. We suggest that the existence of this secondary, active mechanism of SSU transport might be utilized by the cell when a more rapid and directional export is needed

Molecular determinants of nucleosome retention at CpG-rich sequences in mouse spermatozoa

Item does not contain fulltextIn mammalian spermatozoa, most but not all of the genome is densely packaged by protamines. Here we reveal the molecular logic underlying the retention of nucleosomes in mouse spermatozoa, which contain only 1% residual histones. We observe high enrichment throughout the genome of nucleosomes at CpG-rich sequences that lack DNA methylation. Residual nucleosomes are largely composed of the histone H3.3 variant and are trimethylated at Lys4 of histone H3 (H3K4me3). Canonical H3.1 and H3.2 histones are also enriched at CpG-rich promoters marked by Polycomb-mediated H3K27me3, a modification predictive of gene repression in preimplantation embryos. Histone variant-specific nucleosome retention in sperm is strongly associated with nucleosome turnover in round spermatids. Our data show evolutionary conservation of the basic principles of nucleosome retention in mouse and human sperm, supporting a model of epigenetic inheritance by nucleosomes between generations