A Human Gene Coding for a Membrane-associated Nucleic Acid-binding Protein

Siess, Don C.; Vedder, Colleen T.; Merkens, Louise S.; Tanaka, Toshiko; Freed, Alison C.; McCoy, Sharon L.; Heinrich, Michael C.; Deffebach, Mark; Bennett, Robert M.; Hefeneider, Steven H.

doi:10.1074/jbc.m004461200

Cited by 53 publications

(35 citation statements)

References 64 publications

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“…39,40 More recently, a B130 kDa membrane-associated protein with a RING finger domain has also been described with ubiquitous distribution including lymphoid organs. 41 Toll receptor 9 (TLR-9), a transmembrane protein localized at 'endosome-like' cytoplasmatic structures, [42][43][44] mediates the activatory-intracellular signaling of CpG-containing oligonucleotides via interaction with an intracellular MyD88 adapter molecule, IRAK, and TRAF6 45 but not their internalization. Lastly, HLA-class II molecules that are constitutively expressed on B lymphocytes and undergo rapid internalization and recycling upon ligation [46][47][48] could be at play based on the observation that HLA-DR molecules bind oligonucleotides with high affinity (K d 297.9-116.7 nM).…”

Section: Discussionmentioning

confidence: 99%

Spontaneous transgenesis of human B lymphocytes

et al. 2003

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DNA can cross the cell membrane by natural means, but the functional relevance of this phenomenon has not been fully elucidated. Here, we analyzed spontaneous transgenesis of human B cells using plasmid DNA coding for a functional immunoglobulin (Ig) heavy chain gene under the control of a B-cell-specific promoter. Using polymerase chain reaction (PCR), reverse transcriptase-PCR, and flow cytometry in combination, spontaneous transgenesis was documented in Burkitt's lymphoma cell lines, Epstein-Barr virus-transformed cell lines, and peripheral blood B lymphocytes of the mature naïve phenotype (IgM þ /IgD þ /CD27 À ). By immunoelectron microscopy, the internalized DNA was seen in the lysosomes/late endosomes and in the cytosol proximal to the nucleus. Importantly, spontaneously transgenic B cells processed and presented to major histocompatibility complex (MHC)-restricted T lymphocytes a peptide expressed in the transgenic product. This is the first demonstration that primary B lymphocytes possess a program for the spontaneous internalization of DNA, which in turn imparts the cell with new immunological functions. As spontaneous transgenesis is obtained using a nonviral vector, does not require prior cell activation, and is not associated with chromosomal integration, the findings reported here open new possibilities for genetic manipulations of mature naïve B lymphocytes for therapy and vaccination.

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Section: Discussionmentioning

confidence: 99%

Spontaneous transgenesis of human B lymphocytes

et al. 2003

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“…Budker et al hypothesized that naked plasmid DNA is taken up by receptor-mediated pathway by hepatocytes. 29 Certain DNA receptors have been found in various tissues; 30 however, their function has not been elucidated. It has been proposed that the injected DNA solution accumulates mainly in the liver because of its flexible structure, …”

Section: Introductionmentioning

confidence: 99%

Gene Therapy Progress and Prospects: Nonviral vectors

2002

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“…Siess et al (27) attempted expression cloning of a cell surface DNA receptor and obtained a membrane-associated nucleic acid binding protein (MNAB), a 150-kDa protein. The full-length cDNA was obtained from a T cell line, although T cells are not known for efficient DNA uptake.…”

Section: A Macrophage-specific Uptake Receptor?mentioning

confidence: 99%

Differences in Macrophage Activation by Bacterial DNA and CpG-Containing Oligonucleotides

Roberts

Dunn

Terry³

et al. 2005

The Journal of Immunology

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Bacterial DNA activates mouse macrophages, B cells, and dendritic cells in a TLR9-dependent manner. Although short ssCpG-containing phosphodiester oligonucleotides (PO-ODN) can mimic the action of bacterial DNA on macrophages, they are much less immunostimulatory than Escherichia coli DNA. In this study we have assessed the structural differences between E. coli DNA and PO-ODN, which may explain the high activity of bacterial DNA on macrophages. DNA length was found to be the most important variable. Double-strandedness was not responsible for the increased activity of long DNA. DNA adenine methyltransferase (Dam) and DNA cytosine methyltransferase (Dcm) methylation of E. coli DNA did not enhance macrophage NO production. The presence of two CpG motifs on one molecule only marginally improved activity at low concentration, suggesting that ligand-mediated TLR9 cross-linking was not involved. The major contribution was from DNA length. Synthetic ODN >44 nt attained the same levels of activity as bacterial DNA. The response of macrophages to CpG DNA requires endocytic uptake. The length dependence of the CpG ODN response was found to correlate with the presence in macrophages of a length-dependent uptake process for DNA. This transport system was absent from B cells and fibroblasts.

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A Human Gene Coding for a Membrane-associated Nucleic Acid-binding Protein

Cited by 53 publications

References 64 publications

Spontaneous transgenesis of human B lymphocytes

Spontaneous transgenesis of human B lymphocytes

Gene Therapy Progress and Prospects: Nonviral vectors

Differences in Macrophage Activation by Bacterial DNA and CpG-Containing Oligonucleotides

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