1-2-dioleoyloxy-3-(trimethylammonium)propane and Lupus-Erythematosus--Systemic

To examine the effect of transfection of oligodeoxynucleotides (ODNs) containing a CpG motif (CpG-ODN), of which the sequence was derived from circulating DNA in the sera of patients with systemic lupus erythematosus (SLE), on the expression of intercellular adhesion molecule-1 (ICAM-1) and synthesis of mRNA for proinflammatory cytokines and ICAM-1 in human umbilical vein endothelial cells (EC).. A CpG-ODN or a control analogue, GpC-ODN, was transfected into EC. ICAM-1 expression was examined by flow cytometry, and expression of mRNA in EC encoding interleukin 1 (IL1), IL6, IL8, tumour necrosis factor alpha (TNFalpha), interferon gamma (IFNgamma), and ICAM-1 was examined by semiquantitative reverse transcriptase-polymerase chain reaction.. The CpG-ODN augmented the expression of ICAM-1 on EC determined by flow cytometry and increased mRNA levels of IL6, IL8, TNFalpha, IFNgamma, and ICAM-1, but the GpC-ODN did not.. Synthesised DNA, with a sequence corresponding to that of the fragment containing the CpG motif, in sera of patients with SLE was found to enhance ICAM-1 expression on EC, suggesting the participation of circulating DNA fragments in the pathogenesis of vasculitis in SLE.

Topics: Cells, Cultured; CpG Islands; DNA; Endothelium, Vascular; Fatty Acids, Monounsaturated; Flow Cytometry; Fluorescent Dyes; Humans; Intercellular Adhesion Molecule-1; Interferon-gamma; Interleukin-1; Interleukin-6; Interleukin-8; Lupus Erythematosus, Systemic; Quaternary Ammonium Compounds; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Necrosis Factor-alpha

To characterize DNA in sera of patients with systemic lupus erythematosus (SLE) in terms of size, guanine plus cytosine (G+C) content (by percentage), CpG dinucleotide (CpG) (percentage), and effects on mononuclear cells (MNC).. Nine DNA clones were sequenced. Oligodeoxynucleotides with the characteristic CpG motif (TTCGAA or PuPuCGPyPy) were examined for their proliferative effect on MNC by [3H]thymidine incorporation, expression of HLA-DR and intercellular adhesion molecule (ICAM)-1 on monocytes by flow cytometry, and mRNA levels encoding interleukin 12 (IL-12) and interferon-gamma (IFN-gamma) by semiquantitative reverse transcription polymerase chain reaction.. The size of DNA clones ranged from 87 to 318 bp (mean +/- SD, 177+/-68) and enrichment in G+C and CpG ranged from 34.7 to 69.7% (48.1+/-10.7) and 0.63 to 12.8% (4.0+/-4.1), respectively. Three of 9 clones contained the characteristic CpG motif. Oligonucleotides proliferated MNC, and augmented HLA-DR and ICAM-1 expression in company with an increase of mRNA encoding IL-12 and IFN-gamma.. Circulating CpG motif-containing DNA fragments in SLE increased mRNA encoding IL-12 and IFN-gamma, which in turn increased HLA-DR and ICAM-1 on monocytes, resulting in MNC proliferation. This mechanism could contribute to the pathogenesis of SLE.

Topics: Base Sequence; Cell Division; Cells, Cultured; CpG Islands; DNA; Fatty Acids, Monounsaturated; HLA-DR Antigens; Humans; In Vitro Techniques; Intercellular Adhesion Molecule-1; Interferon-gamma; Interleukin-12; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Lymphocyte Activation; Molecular Sequence Data; Oligonucleotide Probes; Quaternary Ammonium Compounds; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger