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Stimulation of Innate and Adaptive Immunity by Using Filamentous Bacteriophage fd Targeted to DEC-205.

D'Apice L, Costa V, Sartorius R, Trovato M, Aprile M, De Berardinis P - J Immunol Res (2015)

Bottom Line: The filamentous bacteriophage fd, codisplaying antigenic determinants and a single chain antibody fragment directed against the dendritic cell receptor DEC-205, is a promising vaccine candidate for its safety and its ability to elicit innate and adaptive immune response in absence of adjuvants.By using a system vaccinology approach based on RNA-Sequencing (RNA-Seq) analysis, we describe a relevant gene modulation in dendritic cells pulsed with anti-DEC-205 bacteriophages fd.RNA-Seq data analysis indicates that the bacteriophage fd virions are sensed as a pathogen by dendritic cells; they activate the danger receptors that trigger an innate immune response and thus confer a strong adjuvanticity that is needed to obtain a long-lasting adaptive immune response.

View Article: PubMed Central - PubMed

Affiliation: Institute of Protein Biochemistry (IBP), National Council of Research, 80131 Naples, Italy.

ABSTRACT
The filamentous bacteriophage fd, codisplaying antigenic determinants and a single chain antibody fragment directed against the dendritic cell receptor DEC-205, is a promising vaccine candidate for its safety and its ability to elicit innate and adaptive immune response in absence of adjuvants. By using a system vaccinology approach based on RNA-Sequencing (RNA-Seq) analysis, we describe a relevant gene modulation in dendritic cells pulsed with anti-DEC-205 bacteriophages fd. RNA-Seq data analysis indicates that the bacteriophage fd virions are sensed as a pathogen by dendritic cells; they activate the danger receptors that trigger an innate immune response and thus confer a strong adjuvanticity that is needed to obtain a long-lasting adaptive immune response.

No MeSH data available.


Related in: MedlinePlus

Real-time PCR validation of RNA seq analysis. The Isg15, Irf7, and Il1b gene expression in BMDCs in vitro challenged with fdsc-αDEC phage particles was measured by real-time PCR. The dashed line corresponds to the mean value of gene expression of PBS treated DCs. Graph shows the fold change (FC) (mean ± SD). ∗∗p < 0.05.
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fig4: Real-time PCR validation of RNA seq analysis. The Isg15, Irf7, and Il1b gene expression in BMDCs in vitro challenged with fdsc-αDEC phage particles was measured by real-time PCR. The dashed line corresponds to the mean value of gene expression of PBS treated DCs. Graph shows the fold change (FC) (mean ± SD). ∗∗p < 0.05.

Mentions: To better dissect our results, we compared them with the already published data [23] describing mouse DCs treated with phylogenetically different organisms, such as bacteria, helminths, and parasites as a paradigm of how DCs undergo marked reprogramming during infection with live pathogens. Our RNAseq data show that in agreement with data obtained with the live pathogens, our procaryotic virus is able to activate specific classes of genes such as the CXCL1 (growth-related oncogene 1 (GRO1)), CXCL2 (GRO2), CCL2, CCL7, and genes encoding the proinflammatory mediators tumor necrosis factor α (TNF-α) and interleukin (IL-1 β) (see Table 1). Moreover, we also found the significant upregulation (more than 19-fold) of CXCL10 (IFN-inducible protein 10); importantly, this chemokine is essential for the generation of protective CD8+ T cell responses and it is produced by dendritic cells following CpG-ODN stimulation [24]. Measuring the expression levels of the antiviral genes Oas3, Oas2, and Eif2ak2 we found that they were 18-, 16-, and 13-fold change increased, respectively, in bacteriophage-treated DCs (Table 1). Finally, expression data revealed a significant upregulation of Interferon-Stimulated Genes (ISG) and in particular of Isg15 gene that was upregulated more than twentyfold, similarly to Irf7 gene (Table 1). The expression of these genes was assessed also by quantitative real-time PCR showing a fold change of 8.6 for Isg15 and 6.6 for Irf7 gene in DC treated with the engineered bacteriophage (Figure 4). Also the Il1b gene expression was measured by real-time PCR and showed a twofold increase of mRNA in fdsc-αDEC treated DC.


Stimulation of Innate and Adaptive Immunity by Using Filamentous Bacteriophage fd Targeted to DEC-205.

D'Apice L, Costa V, Sartorius R, Trovato M, Aprile M, De Berardinis P - J Immunol Res (2015)

Real-time PCR validation of RNA seq analysis. The Isg15, Irf7, and Il1b gene expression in BMDCs in vitro challenged with fdsc-αDEC phage particles was measured by real-time PCR. The dashed line corresponds to the mean value of gene expression of PBS treated DCs. Graph shows the fold change (FC) (mean ± SD). ∗∗p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4563097&req=5

fig4: Real-time PCR validation of RNA seq analysis. The Isg15, Irf7, and Il1b gene expression in BMDCs in vitro challenged with fdsc-αDEC phage particles was measured by real-time PCR. The dashed line corresponds to the mean value of gene expression of PBS treated DCs. Graph shows the fold change (FC) (mean ± SD). ∗∗p < 0.05.
Mentions: To better dissect our results, we compared them with the already published data [23] describing mouse DCs treated with phylogenetically different organisms, such as bacteria, helminths, and parasites as a paradigm of how DCs undergo marked reprogramming during infection with live pathogens. Our RNAseq data show that in agreement with data obtained with the live pathogens, our procaryotic virus is able to activate specific classes of genes such as the CXCL1 (growth-related oncogene 1 (GRO1)), CXCL2 (GRO2), CCL2, CCL7, and genes encoding the proinflammatory mediators tumor necrosis factor α (TNF-α) and interleukin (IL-1 β) (see Table 1). Moreover, we also found the significant upregulation (more than 19-fold) of CXCL10 (IFN-inducible protein 10); importantly, this chemokine is essential for the generation of protective CD8+ T cell responses and it is produced by dendritic cells following CpG-ODN stimulation [24]. Measuring the expression levels of the antiviral genes Oas3, Oas2, and Eif2ak2 we found that they were 18-, 16-, and 13-fold change increased, respectively, in bacteriophage-treated DCs (Table 1). Finally, expression data revealed a significant upregulation of Interferon-Stimulated Genes (ISG) and in particular of Isg15 gene that was upregulated more than twentyfold, similarly to Irf7 gene (Table 1). The expression of these genes was assessed also by quantitative real-time PCR showing a fold change of 8.6 for Isg15 and 6.6 for Irf7 gene in DC treated with the engineered bacteriophage (Figure 4). Also the Il1b gene expression was measured by real-time PCR and showed a twofold increase of mRNA in fdsc-αDEC treated DC.

Bottom Line: The filamentous bacteriophage fd, codisplaying antigenic determinants and a single chain antibody fragment directed against the dendritic cell receptor DEC-205, is a promising vaccine candidate for its safety and its ability to elicit innate and adaptive immune response in absence of adjuvants.By using a system vaccinology approach based on RNA-Sequencing (RNA-Seq) analysis, we describe a relevant gene modulation in dendritic cells pulsed with anti-DEC-205 bacteriophages fd.RNA-Seq data analysis indicates that the bacteriophage fd virions are sensed as a pathogen by dendritic cells; they activate the danger receptors that trigger an innate immune response and thus confer a strong adjuvanticity that is needed to obtain a long-lasting adaptive immune response.

View Article: PubMed Central - PubMed

Affiliation: Institute of Protein Biochemistry (IBP), National Council of Research, 80131 Naples, Italy.

ABSTRACT
The filamentous bacteriophage fd, codisplaying antigenic determinants and a single chain antibody fragment directed against the dendritic cell receptor DEC-205, is a promising vaccine candidate for its safety and its ability to elicit innate and adaptive immune response in absence of adjuvants. By using a system vaccinology approach based on RNA-Sequencing (RNA-Seq) analysis, we describe a relevant gene modulation in dendritic cells pulsed with anti-DEC-205 bacteriophages fd. RNA-Seq data analysis indicates that the bacteriophage fd virions are sensed as a pathogen by dendritic cells; they activate the danger receptors that trigger an innate immune response and thus confer a strong adjuvanticity that is needed to obtain a long-lasting adaptive immune response.

No MeSH data available.


Related in: MedlinePlus