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Cytokine response to the RSV antigen delivered by dendritic cell-directed vaccination in congenic chicken lines

View Article: PubMed Central - PubMed

ABSTRACT

Systems of antigen delivery into antigen-presenting cells represent an important novel strategy in chicken vaccine development. In this study, we verified the ability of Rous sarcoma virus (RSV) antigens fused with streptavidin to be targeted by specific biotinylated monoclonal antibody (anti-CD205) into dendritic cells and induce virus-specific protective immunity. The method was tested in four congenic lines of chickens that are either resistant or susceptible to the progressive growth of RSV-induced tumors. Our analyses confirmed that the biot-anti-CD205-SA-FITC complex was internalized by chicken splenocytes. In the cytokine expression profile, several significant differences were evident between RSV-challenged progressor and regressor chicken lines. A significant up-regulation of IL-2, IL-12, IL-15, and IL-18 expression was detected in immunized chickens of both regressor and progressor groups. Of these cytokines, IL-2 and IL-12 were most up-regulated 14 days post-challenge (dpc), while IL-15 and IL-18 were most up-regulated at 28 dpc. On the contrary, IL-10 expression was significantly down-regulated in all immunized groups of progressor chickens at 14 dpc. We detected significant up-regulation of IL-17 in the group of immunized progressors. LITAF down-regulation with iNOS up-regulation was especially observed in the progressor group of immunized chickens that developed large tumors. Based on the increased expression of cytokines specific for activated dendritic cells, we conclude that our system is able to induce partial stimulation of specific cell types involved in cell-mediated immunity.

Electronic supplementary material: The online version of this article (doi:10.1186/s13567-017-0423-8) contains supplementary material, which is available to authorized users.

No MeSH data available.


In vitro Ag restimulation. The v-src-specific interleukin (IL-12, IL-2, IFNγ, IL-10) responses were measured in chickens of the CB and CB.RI lines using the Bio-Plex system in the culture supernatants. Horizontal bars indicate the mean values. The results represent at least two independent experiments. Control (n = 5 + 5): non-restimulated control; SA-vsrc-10 (n = 5 + 5): samples restimulated with the mixture of tetramers SA-vsrcA and SA-vsrcB of immunized and non-immunized animals; SA-10 (n = 5 + 5): free SA restimulation control. Statistical significance determined by the Student’s t test: *p < 0.05; **p < 0.01; ***p < 0.001.
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Fig3: In vitro Ag restimulation. The v-src-specific interleukin (IL-12, IL-2, IFNγ, IL-10) responses were measured in chickens of the CB and CB.RI lines using the Bio-Plex system in the culture supernatants. Horizontal bars indicate the mean values. The results represent at least two independent experiments. Control (n = 5 + 5): non-restimulated control; SA-vsrc-10 (n = 5 + 5): samples restimulated with the mixture of tetramers SA-vsrcA and SA-vsrcB of immunized and non-immunized animals; SA-10 (n = 5 + 5): free SA restimulation control. Statistical significance determined by the Student’s t test: *p < 0.05; **p < 0.01; ***p < 0.001.

Mentions: In order to verify the changed expression of the three major cytokines (IL-2, IL-12 and IFNγ), the in vitro restimulation of PBMC was performed. The levels of three cytokines detected in PBMC culture supernatants (Il-2, IL-12 and IFNγ) were significantly increased in the immunized groups that were restimulated with v-src antigen. On the contrary, the amount of IL-10 was at the same level as in PBMC restimulated with v-src or control antigen or in the group without restimulation respectively (Figure 3).Figure 3


Cytokine response to the RSV antigen delivered by dendritic cell-directed vaccination in congenic chicken lines
In vitro Ag restimulation. The v-src-specific interleukin (IL-12, IL-2, IFNγ, IL-10) responses were measured in chickens of the CB and CB.RI lines using the Bio-Plex system in the culture supernatants. Horizontal bars indicate the mean values. The results represent at least two independent experiments. Control (n = 5 + 5): non-restimulated control; SA-vsrc-10 (n = 5 + 5): samples restimulated with the mixture of tetramers SA-vsrcA and SA-vsrcB of immunized and non-immunized animals; SA-10 (n = 5 + 5): free SA restimulation control. Statistical significance determined by the Student’s t test: *p < 0.05; **p < 0.01; ***p < 0.001.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5382389&req=5

Fig3: In vitro Ag restimulation. The v-src-specific interleukin (IL-12, IL-2, IFNγ, IL-10) responses were measured in chickens of the CB and CB.RI lines using the Bio-Plex system in the culture supernatants. Horizontal bars indicate the mean values. The results represent at least two independent experiments. Control (n = 5 + 5): non-restimulated control; SA-vsrc-10 (n = 5 + 5): samples restimulated with the mixture of tetramers SA-vsrcA and SA-vsrcB of immunized and non-immunized animals; SA-10 (n = 5 + 5): free SA restimulation control. Statistical significance determined by the Student’s t test: *p < 0.05; **p < 0.01; ***p < 0.001.
Mentions: In order to verify the changed expression of the three major cytokines (IL-2, IL-12 and IFNγ), the in vitro restimulation of PBMC was performed. The levels of three cytokines detected in PBMC culture supernatants (Il-2, IL-12 and IFNγ) were significantly increased in the immunized groups that were restimulated with v-src antigen. On the contrary, the amount of IL-10 was at the same level as in PBMC restimulated with v-src or control antigen or in the group without restimulation respectively (Figure 3).Figure 3

View Article: PubMed Central - PubMed

ABSTRACT

Systems of antigen delivery into antigen-presenting cells represent an important novel strategy in chicken vaccine development. In this study, we verified the ability of Rous sarcoma virus (RSV) antigens fused with streptavidin to be targeted by specific biotinylated monoclonal antibody (anti-CD205) into dendritic cells and induce virus-specific protective immunity. The method was tested in four congenic lines of chickens that are either resistant or susceptible to the progressive growth of RSV-induced tumors. Our analyses confirmed that the biot-anti-CD205-SA-FITC complex was internalized by chicken splenocytes. In the cytokine expression profile, several significant differences were evident between RSV-challenged progressor and regressor chicken lines. A significant up-regulation of IL-2, IL-12, IL-15, and IL-18 expression was detected in immunized chickens of both regressor and progressor groups. Of these cytokines, IL-2 and IL-12 were most up-regulated 14&nbsp;days post-challenge (dpc), while IL-15 and IL-18 were most up-regulated at 28&nbsp;dpc. On the contrary, IL-10 expression was significantly down-regulated in all immunized groups of progressor chickens at 14&nbsp;dpc. We detected significant up-regulation of IL-17 in the group of immunized progressors. LITAF down-regulation with iNOS up-regulation was especially observed in the progressor group of immunized chickens that developed large tumors. Based on the increased expression of cytokines specific for activated dendritic cells, we conclude that our system is able to induce partial stimulation of specific cell types involved in cell-mediated immunity.

Electronic supplementary material: The online version of this article (doi:10.1186/s13567-017-0423-8) contains supplementary material, which is available to authorized users.

No MeSH data available.