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Antigen delivery by filamentous bacteriophage fd displaying an anti-DEC-205 single-chain variable fragment confers adjuvanticity by triggering a TLR9-mediated immune response.

Sartorius R, D'Apice L, Trovato M, Cuccaro F, Costa V, De Leo MG, Marzullo VM, Biondo C, D'Auria S, De Matteis MA, Ciccodicola A, De Berardinis P - EMBO Mol Med (2015)

Bottom Line: A significant differential expression of genes involved in innate immunity, co-stimulation and cytokine production was observed.We also found that fdsc-αDEC is delivered into LAMP-1-positive compartments and co-localizes with TLR9.Thus, phage particles containing a single-strand DNA genome rich in CpG motifs delivered via DEC-205 are able to intercept and trigger the active TLR9 innate immune receptor into late endosome/lysosomes and to enhance the immunogenicity of the displayed antigenic determinants.

View Article: PubMed Central - PubMed

Affiliation: Institute of Protein Biochemistry, National Council of Research, Naples, Italy.

No MeSH data available.


Related in: MedlinePlus

fdsc-αDEC localizes with TLR9 in DCsA–D BMDCs from C57BL/6 mice were stably transduced with YFP-tagged TLR9. Cells grown on coverslips were then incubated with TRITC-conjugated wild-type (A, B) or fdsc-αDEC (C, D) bacteriophages for 6 h. Cells were then washed, fixed and analyzed by confocal microscopy. Nuclei were counterstained with Hoechst. (B, D) are enlargements of a single cell. Representative images are shown. Numbers at bottom of right panels indicate the percentage of TLR9 bacteriophage co-localization (mean values ± SD, N = 2, n = 100). Scale bars, 10 mm.
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fig05: fdsc-αDEC localizes with TLR9 in DCsA–D BMDCs from C57BL/6 mice were stably transduced with YFP-tagged TLR9. Cells grown on coverslips were then incubated with TRITC-conjugated wild-type (A, B) or fdsc-αDEC (C, D) bacteriophages for 6 h. Cells were then washed, fixed and analyzed by confocal microscopy. Nuclei were counterstained with Hoechst. (B, D) are enlargements of a single cell. Representative images are shown. Numbers at bottom of right panels indicate the percentage of TLR9 bacteriophage co-localization (mean values ± SD, N = 2, n = 100). Scale bars, 10 mm.

Mentions: To verify the co-localization of phage particles internalized via the DEC-205 receptor with TLR9 in dendritic cells, we expressed recombinant TLR9 C-terminally fused to yellow fluorescent protein (YFP) in BMDCs. Using TRITC-conjugated bacteriophages and confocal microscopy, we were able to assess the co-localization of phage particles internalized by DCs with the TLR9-YFP. We found that fdsc-αDEC particles, which are able to activate DCs, induce a high number of TLR9-positive structures compared to cells incubated with fd wild-type particles (Fig5A and C, left panels). Moreover, the majority (82.3%) of the fdsc-αDEC virions co-localized with TLR9 (Fig5C and D) compared to only 4.6% co-localization of the fd wild-type bacteriophages (Fig5A and B).


Antigen delivery by filamentous bacteriophage fd displaying an anti-DEC-205 single-chain variable fragment confers adjuvanticity by triggering a TLR9-mediated immune response.

Sartorius R, D'Apice L, Trovato M, Cuccaro F, Costa V, De Leo MG, Marzullo VM, Biondo C, D'Auria S, De Matteis MA, Ciccodicola A, De Berardinis P - EMBO Mol Med (2015)

fdsc-αDEC localizes with TLR9 in DCsA–D BMDCs from C57BL/6 mice were stably transduced with YFP-tagged TLR9. Cells grown on coverslips were then incubated with TRITC-conjugated wild-type (A, B) or fdsc-αDEC (C, D) bacteriophages for 6 h. Cells were then washed, fixed and analyzed by confocal microscopy. Nuclei were counterstained with Hoechst. (B, D) are enlargements of a single cell. Representative images are shown. Numbers at bottom of right panels indicate the percentage of TLR9 bacteriophage co-localization (mean values ± SD, N = 2, n = 100). Scale bars, 10 mm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4520660&req=5

fig05: fdsc-αDEC localizes with TLR9 in DCsA–D BMDCs from C57BL/6 mice were stably transduced with YFP-tagged TLR9. Cells grown on coverslips were then incubated with TRITC-conjugated wild-type (A, B) or fdsc-αDEC (C, D) bacteriophages for 6 h. Cells were then washed, fixed and analyzed by confocal microscopy. Nuclei were counterstained with Hoechst. (B, D) are enlargements of a single cell. Representative images are shown. Numbers at bottom of right panels indicate the percentage of TLR9 bacteriophage co-localization (mean values ± SD, N = 2, n = 100). Scale bars, 10 mm.
Mentions: To verify the co-localization of phage particles internalized via the DEC-205 receptor with TLR9 in dendritic cells, we expressed recombinant TLR9 C-terminally fused to yellow fluorescent protein (YFP) in BMDCs. Using TRITC-conjugated bacteriophages and confocal microscopy, we were able to assess the co-localization of phage particles internalized by DCs with the TLR9-YFP. We found that fdsc-αDEC particles, which are able to activate DCs, induce a high number of TLR9-positive structures compared to cells incubated with fd wild-type particles (Fig5A and C, left panels). Moreover, the majority (82.3%) of the fdsc-αDEC virions co-localized with TLR9 (Fig5C and D) compared to only 4.6% co-localization of the fd wild-type bacteriophages (Fig5A and B).

Bottom Line: A significant differential expression of genes involved in innate immunity, co-stimulation and cytokine production was observed.We also found that fdsc-αDEC is delivered into LAMP-1-positive compartments and co-localizes with TLR9.Thus, phage particles containing a single-strand DNA genome rich in CpG motifs delivered via DEC-205 are able to intercept and trigger the active TLR9 innate immune receptor into late endosome/lysosomes and to enhance the immunogenicity of the displayed antigenic determinants.

View Article: PubMed Central - PubMed

Affiliation: Institute of Protein Biochemistry, National Council of Research, Naples, Italy.

No MeSH data available.


Related in: MedlinePlus