Double-stranded deoxyribonucleic acid binds to HLA class II molecules and inhibits HLA class II-mediated antigen presentation

Filaci, Gilberto; Contini, Paola; Grasso, I.; Bignardi, Donatella; Ghio, Massimo; Lanza, Lorella; Scudeletti, Marco; Puppo, Francesco; Bolognesi, M.; Accolla, Roberto S.; Indiveri, Francesco

doi:10.1002/(sici)1521-4141(199812)28:12<3968::aid-immu3968>3.0.co;2-6

Cited by 12 publications

(8 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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). 49 Although attractive, our evidence in favor of this hypothesis is modest. For instance, DNA internalization was diminished but not abrogated in HLA class II competent (Raji) cells as compared to HLA class II deficient (RJ 2.2.1) cells (not shown).…”

Section: Discussionmentioning

confidence: 78%

Spontaneous transgenesis of human B lymphocytes

et al. 2003

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionmentioning

confidence: 78%

Spontaneous transgenesis of human B lymphocytes

et al. 2003

View full text Add to dashboard Cite

show abstract

“…However, because PTO‐modification conveys binding affinity to many proteins, it is unclear whether binding of PTO‐ODN to certain proteins is really linked to structures involved in uptake. In fact, an uptake function for ODN has been attributed to many distinct cell surface structures: surface immunoglobulins, 38 human leucocyte antigen (HLA) class II molecules 39 and Mac‐1 (CD11b/CD18) 37 . However, Mac‐1 −/− knockout mice showed no alterations in responses to CpG‐ODN 40 .…”

Section: Discussionmentioning

confidence: 99%

Phosphodiester CpG oligonucleotides as adjuvants: polyguanosine runs enhance cellular uptake and improve immunostimulative activity of phosphodiester CpG oligonucleotides in vitro and in vivo

et al. 2002

View full text Add to dashboard Cite

SUMMARYBacterial DNA and oligonucleotides (ODN) containing CpG-motifs strongly activate cells of the immune system. Accordingly CpG-DNA is a powerful adjuvant in vaccination protocols for B-cell as well as for cytotoxic T-cell responses. A decisive propensity of CpG-DNA is its capacity to induce preferentially T helper type 1 (Th1)-dominated immune responses. To exert its function CpG-DNA has to be taken up by responsive cells, e.g. antigen-presenting cells (APC). The rate of uptake is influenced by the DNA's backbone modification and critically determines activity of CpG-DNA. CpG ODN with a phosphothioate backbone (PTO) are currently used for most in vivo and in vitro studies, since PTO modification protects ODN from the attack of nucleases. However, after administration of PTO-modified CpG-ODN long-lasting effects including lymphadenopathy as well as sustained local interferon-c (IFN-c) and interleukin-12 (IL-12) production have been reported. To circumvent these restrictions we investigated the effects of DNA sequence as well as DNA backbone modification on cellular uptake and resulting immunostimulation. We show here that uptake of phosphodiester (PO)-CpG-ODN can be strongly enhanced by poly guanosine runs added at the 3k end of the ODN. In addition these ODN showed an improved immunostimulatory activity in vivo and in vitro. This included protection of mice against lethal Th2-dependent leishmaniasis as well as priming of antigen specific Th1 responses. More importantly, guanosine-rich PO-CpG-ODN neither induced lymphadenopathy nor prolonged cytokine production after local administration. Since these improved PO ODN are efficient in vitro and in vivo and lack long lasting undesired effects they could be used preferably as adjuvants in vaccination protocols.

show abstract

“…Of importance, dsDNA or ODNs can bind HLA class II molecules specifically, and dsDNA or ODNs bound HLA-II ϩ Raji cells, but not HLA-II Ϫ RJ2.2.5 cells. Moreover, the mixed lymphocyte reaction and Ag-specific T cell proliferation were inhibited by preincubation of stimulator cells or Ag-specific T cell lines or anti-CD3 mAb-stimulated human PBMC with dsDNA (28). These results suggest that dsDNA can bind HLA class II molecules and can inhibit Ag presentation.…”

Section: Discussionmentioning

confidence: 99%

The Role of Cell Surface Receptors in the Activation of Human B Cells by Phosphorothioate Oligonucleotides

Liang

Reich

Pisetsky

et al. 2000

The Journal of Immunology

View full text Add to dashboard Cite

Phosphorothioate oligodeoxynucleotides (sODN) containing the CpG motif or TCG repeats induce T cell-independent polyclonal activation of human B cells. To elucidate the mechanism of this response, the role of cell surface receptors was investigated. Sepharose beads coated with stimulatory but not nonstimulatory sODNs induced B cell proliferation comparably with soluble sODNs. The B cell stimulatory activity of Sepharose-bound sODN did not result from free sODN released from the beads since media incubated with coated beads were inactive. Using FITC-labeled sODNs as probes, binding to human B cells could be detected by flow cytometry. Binding was rapid, saturable, initially temperature independent, but with a rapid off-rate. Competition studies indicated that both stimulatory sODNs and minimally stimulatory sODNs bound to the same receptor. By contrast, phosphodiester oligonucleotides with the same nucleotide sequence as sODNs and bacterial DNA inhibited the binding of sODNs to B cells minimally. Charge appeared to contribute to the binding of sODNs to B cells since binding of sODNs was competitively inhibited by negatively charged molecules, including fucoidan, poly I, and polyvinyl sulfate. These data indicate that human B cells bind sODNs by a receptor-mediated mechanism that is necessary but not sufficient for polyclonal activation.

show abstract

Double-stranded deoxyribonucleic acid binds to HLA class II molecules and inhibits HLA class II-mediated antigen presentation

Cited by 12 publications

References 30 publications

Spontaneous transgenesis of human B lymphocytes

Spontaneous transgenesis of human B lymphocytes

Phosphodiester CpG oligonucleotides as adjuvants: polyguanosine runs enhance cellular uptake and improve immunostimulative activity of phosphodiester CpG oligonucleotides in vitro and in vivo

The Role of Cell Surface Receptors in the Activation of Human B Cells by Phosphorothioate Oligonucleotides

Contact Info

Product

Resources

About