Alijagic A et al. (2021), Immunomodulatory Function of Polyvinylpyrrolido...

ECB-ART-48899

Nanomaterials (Basel) 2021 Oct 08;1110:. doi: 10.3390/nano11102646.

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Immunomodulatory Function of Polyvinylpyrrolidone (PVP)-Functionalized Gold Nanoparticles in Vibrio-Stimulated Sea Urchin Immune Cells.

Alijagic A , Bonura A , Barbero F , Puntes VF , Gervasi F , Pinsino A .

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We investigated the role of the gold nanoparticles functionalized with polyvinylpyrrolidone (PVP-AuNPs) on the innate immune response against an acute infection caused by Vibrio anguillarum in an in vitro immunological nonmammalian next-generation model, the sea urchin Paracentrotus lividus. To profile the immunomodulatory function of PVP-AuNPs (0.1 μg mL-1) in sea urchin immune cells stimulated by Vibrio (10 μg mL-1) for 3 h, we focused on the baseline immunological state of the donor, and we analysed the topography, cellular metabolism, and expression of human cell surface antigens of the exposed cells, as well as the signalling leading the interaction between PVP-AuNPs and the Vibrio-stimulated cells. PVP-AuNPs are not able to silence the inflammatory signalling (TLR4/p38MAPK/NF-κB signalling) that involves the whole population of P. lividus immune cells exposed to Vibrio. However, our findings emphasise the ability of PVP-AuNPs to stimulate a subset of rare cells (defined here as Group 3) that express CD45 and CD14 antigens on their surface, which are known to be involved in immune cell maturation and macrophage activation in humans. Our evidence on how PVP-AuNPs may stimulate sea urchin immune cells represents an important starting point for planning new research work on the topic.

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	Figure 1. PVP–AuNP behaviour in the sea urchin culture media. (A) SEM image of PVP–AuNPs (upper panel); DLS size distribution of PVP–AuNPs (10 µg mL−1) dispersed in CCM at 3 h of exposure (lower panel). (B) UV-vis spectra of PVP–AuNPs (0.2, 1, 10 μg mL−1) dispersed in CCM at 0 (red line) and 3 h (blue line) of exposure, and in mQ water (green line). (C) PVP–AuNPs dispersed in cell-free coelomic fluid (CF) plus CCM at 0.2, 1, and 10 μg mL−1 and 0 and 3 h of exposure (red line and blue line, respectively). (D) PVP–AuNPs dispersed in CF cell-free plus CCM at 0.2 μg mL−1 and 0 and 3 h of exposure (red line and blue line, respectively).
	Figure 2. Sea urchin immune cells under in vitro exposure. (A) Short-term housed P. lividus (2–4 weeks). (B) Macroscopic overview of the immune cells from short-term housed sea urchins after a few seconds of exposure (V. anguillarum, PVP–AuNPs, Vibrio plus PVP–AuNPs). (C) Sea urchin immune cells from short-term housed sea urchin after 3 h of exposure in vitro. (D) Long-term housed P. lividus (6–8 months). (E) Macroscopic overview of the immune cells from long-term housed sea urchins after a few seconds of exposure (V. anguillarum, PVP–AuNPs, Vibrio plus PVP–AuNPs). (F) Sea urchin immune cells from long-term housed sea urchin after 3 h of exposure in vitro.
	Figure 3. Real-time viability assay reveals how the baseline immunological state interferes with the capability of the cells to sense the infection. The cellular viability/metabolism of the immune cells exposed to PVP–AuNPs (0.1 μg mL−1), Vibrio anguillarum (10 μg mL−1), and particle–bacteria combination was measured in cells from both short-term housed donors (A) and long-term housed donors (B). The number of donors was 12 short-term and 5 long-term housed sea urchins. Levels are expressed in both relative luminescence unit (RLU) (on the left), and arbitrary units (fold increase or decrease compared to controls set to 1) (dotted line) (on the right). Data are reported as the mean ± SD; asterisks () indicate significant differences among groups (** p < 0.001).
	Figure 4. Two-parameter flow cytometric analysis of immune cells of an immunologically quiescent sea urchin Paracentrotus lividus via forward scatter (FSC) and sideward scatter (SSC). (A) Forward/side scatter profile of total immune cells harvested from the donor in the coelomic calcium medium (CCM, anticoagulant solution): unexposed cells (control), and cells exposed to PVP–AuNPs (0.1 μg mL−1), Vibrio (10 μg mL−1), and particle–bacteria combination. (B) Forward/side scatter profile of total immune cells harvested from the same donor without CCM. The gating of distinct populations is shown, and the percentage of cells in each of these gates is provided.
	Figure 5. PVP–AuNPs are not able to block the powerful inflammatory response involving the TLR4/NF-κB signalling pathway activated by Vibrio. Representative immunoblotting shows results for the selected subset of proteins in cells exposed to PVP–AuNPs (0.1 μg mL−1), Vibrio (10 μg mL−1) and particles in combination with bacteria for 3 h. Histograms are representative of the means ± SD of six replicates after normalisation with actin. Protein levels are reported in arbitrary units (fold increase or decrease compared to controls that are set to 1). Here, a representative image of the protein levels obtained from three individual donors per group is reported. * p < 0.05, p < 0.01, * p < 0.001.

References [+] :

Alijagic, Gold nanoparticles coated with polyvinylpyrrolidone and sea urchin extracellular molecules induce transient immune activation. 2021, Pubmed, Echinobase