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Efficient uptake of blood-borne BK and JC polyomavirus-like particles in endothelial cells of liver sinusoids and renal vasa recta.

Simon-Santamaria J, Rinaldo CH, Kardas P, Li R, Malovic I, Elvevold K, McCourt P, Smedsrød B, Hirsch HH, Sørensen KK - PLoS ONE (2014)

Bottom Line: Most VLP-positive vessels in renal medulla did not express PV-1/Meca 32, suggesting location to the non-fenestrated part of vasa recta.The endothelial cells of these vessels also efficiently endocytosed a scavenger receptor ligand, formaldehyde-denatured albumin, suggesting high endocytic activity compared to other renal endothelia.We conclude that LSECs very effectively cleared a large fraction of blood-borne BK- and JC-VLPs, indicating a central role of these cells in early removal of polyomavirus from the circulation.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biology, UiT - The Arctic University of Norway, Tromsø, Norway.

ABSTRACT
Liver sinusoidal endothelial cells (LSECs) are specialized scavenger cells that mediate high-capacity clearance of soluble waste macromolecules and colloid material, including blood-borne adenovirus. To explore if LSECs function as a sink for other viruses in blood, we studied the fate of virus-like particles (VLPs) of two ubiquitous human DNA viruses, BK and JC polyomavirus, in mice. Like complete virions, VLPs specifically bind to receptors and enter cells, but unlike complete virions, they cannot replicate. 125I-labeled VLPs were used to assess blood decay, organ-, and hepatocellular distribution of ligand, and non-labeled VLPs to examine cellular uptake by immunohisto- and -cytochemistry. BK- and JC-VLPs rapidly distributed to liver, with lesser uptake in kidney and spleen. Liver uptake was predominantly in LSECs. Blood half-life (∼1 min), and tissue distribution of JC-VLPs and two JC-VLP-mutants (L55F and S269F) that lack sialic acid binding affinity, were similar, indicating involvement of non-sialic acid receptors in cellular uptake. Liver uptake was not mediated by scavenger receptors. In spleen, the VLPs localized to the red pulp marginal zone reticuloendothelium, and in kidney to the endothelial lining of vasa recta segments, and the transitional epithelium of renal pelvis. Most VLP-positive vessels in renal medulla did not express PV-1/Meca 32, suggesting location to the non-fenestrated part of vasa recta. The endothelial cells of these vessels also efficiently endocytosed a scavenger receptor ligand, formaldehyde-denatured albumin, suggesting high endocytic activity compared to other renal endothelia. We conclude that LSECs very effectively cleared a large fraction of blood-borne BK- and JC-VLPs, indicating a central role of these cells in early removal of polyomavirus from the circulation. In addition, we report the novel finding that a subpopulation of endothelial cells in kidney, the main organ of polyomavirus persistence, showed selective and rapid uptake of VLPs, suggesting a role in viremic organ tropism.

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JC-VLP uptake in spleen.Panels A–I: Mouse tissues were perfusion fixed 7 min after intravenous injection of JC-VLPs. Paraffin sections of spleen were double immune labeled using a rabbit antiserum against BK-VP1 [53] (reacts also with JC-VP1), and antibodies against F4/80 (A–C), CD31 (D–F) or CD163 (G–I). Antibodies are listed in Table 1. The VP1 staining pattern (green) showed that the uptake of VLPs was concentrated in the red pulp marginal zone, mz (A), and here partly co-localized with F4/80 (C, arrow), and CD31 (F, arrows), but not with CD163 (I; arrow indicates VP1, and arrow head to CD163 staining). The F4/80 (B, C), CD31 (E, F), and CD163 (H, I) staining patterns are all shown in red. Panels J–L: In J–L, tail vein injection of JC-VLPs was followed by injection of FITC-FSA in the opposite tail vein and tissues perfusion fixed 15 min after the VLP administration. Both JC-VLP (K; red fluorescence) and FITC-FSA (J; green fluorescence) distributed to the reticuloendothelial network of the spleen red pulp marginal zone (L; arrows point to overlap of JC-VLP staining and FITC-FSA uptake).
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pone-0111762-g009: JC-VLP uptake in spleen.Panels A–I: Mouse tissues were perfusion fixed 7 min after intravenous injection of JC-VLPs. Paraffin sections of spleen were double immune labeled using a rabbit antiserum against BK-VP1 [53] (reacts also with JC-VP1), and antibodies against F4/80 (A–C), CD31 (D–F) or CD163 (G–I). Antibodies are listed in Table 1. The VP1 staining pattern (green) showed that the uptake of VLPs was concentrated in the red pulp marginal zone, mz (A), and here partly co-localized with F4/80 (C, arrow), and CD31 (F, arrows), but not with CD163 (I; arrow indicates VP1, and arrow head to CD163 staining). The F4/80 (B, C), CD31 (E, F), and CD163 (H, I) staining patterns are all shown in red. Panels J–L: In J–L, tail vein injection of JC-VLPs was followed by injection of FITC-FSA in the opposite tail vein and tissues perfusion fixed 15 min after the VLP administration. Both JC-VLP (K; red fluorescence) and FITC-FSA (J; green fluorescence) distributed to the reticuloendothelial network of the spleen red pulp marginal zone (L; arrows point to overlap of JC-VLP staining and FITC-FSA uptake).

Mentions: In spleen, the BK- and JC-VLPs distributed to the marginal zone that separates the lymphocyte rich white pulp from the red pulp (Figure 9, and Figure S6). The VP1 labeling showed a net-like staining pattern in this region. Double immune labeling for VP1 and the macrophage marker F4/80 (Figure 9A–C), or CD31 (Figure 9D–F), which is expressed by many endothelial cells, revealed scattered co-localisation of VLPs with CD31 and F4/80 positive cells, suggesting that the marginal zone reticuloendothelial network of macrophages and endothelial cells are involved in VLP uptake in the spleen. Staining for the macrophage antigen CD163 [63] did not co-localize with VP1 staining (Figure 9G–I), indicating VLP uptake only in subsets of spleen red pulp macrophages. Some of the JC-VLP uptake in spleen co-localized with FITC-FSA which also distributed to the reticuloendothelium of the spleen red pulp marginal zone (Figure 9J–L).


Efficient uptake of blood-borne BK and JC polyomavirus-like particles in endothelial cells of liver sinusoids and renal vasa recta.

Simon-Santamaria J, Rinaldo CH, Kardas P, Li R, Malovic I, Elvevold K, McCourt P, Smedsrød B, Hirsch HH, Sørensen KK - PLoS ONE (2014)

JC-VLP uptake in spleen.Panels A–I: Mouse tissues were perfusion fixed 7 min after intravenous injection of JC-VLPs. Paraffin sections of spleen were double immune labeled using a rabbit antiserum against BK-VP1 [53] (reacts also with JC-VP1), and antibodies against F4/80 (A–C), CD31 (D–F) or CD163 (G–I). Antibodies are listed in Table 1. The VP1 staining pattern (green) showed that the uptake of VLPs was concentrated in the red pulp marginal zone, mz (A), and here partly co-localized with F4/80 (C, arrow), and CD31 (F, arrows), but not with CD163 (I; arrow indicates VP1, and arrow head to CD163 staining). The F4/80 (B, C), CD31 (E, F), and CD163 (H, I) staining patterns are all shown in red. Panels J–L: In J–L, tail vein injection of JC-VLPs was followed by injection of FITC-FSA in the opposite tail vein and tissues perfusion fixed 15 min after the VLP administration. Both JC-VLP (K; red fluorescence) and FITC-FSA (J; green fluorescence) distributed to the reticuloendothelial network of the spleen red pulp marginal zone (L; arrows point to overlap of JC-VLP staining and FITC-FSA uptake).
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pone-0111762-g009: JC-VLP uptake in spleen.Panels A–I: Mouse tissues were perfusion fixed 7 min after intravenous injection of JC-VLPs. Paraffin sections of spleen were double immune labeled using a rabbit antiserum against BK-VP1 [53] (reacts also with JC-VP1), and antibodies against F4/80 (A–C), CD31 (D–F) or CD163 (G–I). Antibodies are listed in Table 1. The VP1 staining pattern (green) showed that the uptake of VLPs was concentrated in the red pulp marginal zone, mz (A), and here partly co-localized with F4/80 (C, arrow), and CD31 (F, arrows), but not with CD163 (I; arrow indicates VP1, and arrow head to CD163 staining). The F4/80 (B, C), CD31 (E, F), and CD163 (H, I) staining patterns are all shown in red. Panels J–L: In J–L, tail vein injection of JC-VLPs was followed by injection of FITC-FSA in the opposite tail vein and tissues perfusion fixed 15 min after the VLP administration. Both JC-VLP (K; red fluorescence) and FITC-FSA (J; green fluorescence) distributed to the reticuloendothelial network of the spleen red pulp marginal zone (L; arrows point to overlap of JC-VLP staining and FITC-FSA uptake).
Mentions: In spleen, the BK- and JC-VLPs distributed to the marginal zone that separates the lymphocyte rich white pulp from the red pulp (Figure 9, and Figure S6). The VP1 labeling showed a net-like staining pattern in this region. Double immune labeling for VP1 and the macrophage marker F4/80 (Figure 9A–C), or CD31 (Figure 9D–F), which is expressed by many endothelial cells, revealed scattered co-localisation of VLPs with CD31 and F4/80 positive cells, suggesting that the marginal zone reticuloendothelial network of macrophages and endothelial cells are involved in VLP uptake in the spleen. Staining for the macrophage antigen CD163 [63] did not co-localize with VP1 staining (Figure 9G–I), indicating VLP uptake only in subsets of spleen red pulp macrophages. Some of the JC-VLP uptake in spleen co-localized with FITC-FSA which also distributed to the reticuloendothelium of the spleen red pulp marginal zone (Figure 9J–L).

Bottom Line: Most VLP-positive vessels in renal medulla did not express PV-1/Meca 32, suggesting location to the non-fenestrated part of vasa recta.The endothelial cells of these vessels also efficiently endocytosed a scavenger receptor ligand, formaldehyde-denatured albumin, suggesting high endocytic activity compared to other renal endothelia.We conclude that LSECs very effectively cleared a large fraction of blood-borne BK- and JC-VLPs, indicating a central role of these cells in early removal of polyomavirus from the circulation.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biology, UiT - The Arctic University of Norway, Tromsø, Norway.

ABSTRACT
Liver sinusoidal endothelial cells (LSECs) are specialized scavenger cells that mediate high-capacity clearance of soluble waste macromolecules and colloid material, including blood-borne adenovirus. To explore if LSECs function as a sink for other viruses in blood, we studied the fate of virus-like particles (VLPs) of two ubiquitous human DNA viruses, BK and JC polyomavirus, in mice. Like complete virions, VLPs specifically bind to receptors and enter cells, but unlike complete virions, they cannot replicate. 125I-labeled VLPs were used to assess blood decay, organ-, and hepatocellular distribution of ligand, and non-labeled VLPs to examine cellular uptake by immunohisto- and -cytochemistry. BK- and JC-VLPs rapidly distributed to liver, with lesser uptake in kidney and spleen. Liver uptake was predominantly in LSECs. Blood half-life (∼1 min), and tissue distribution of JC-VLPs and two JC-VLP-mutants (L55F and S269F) that lack sialic acid binding affinity, were similar, indicating involvement of non-sialic acid receptors in cellular uptake. Liver uptake was not mediated by scavenger receptors. In spleen, the VLPs localized to the red pulp marginal zone reticuloendothelium, and in kidney to the endothelial lining of vasa recta segments, and the transitional epithelium of renal pelvis. Most VLP-positive vessels in renal medulla did not express PV-1/Meca 32, suggesting location to the non-fenestrated part of vasa recta. The endothelial cells of these vessels also efficiently endocytosed a scavenger receptor ligand, formaldehyde-denatured albumin, suggesting high endocytic activity compared to other renal endothelia. We conclude that LSECs very effectively cleared a large fraction of blood-borne BK- and JC-VLPs, indicating a central role of these cells in early removal of polyomavirus from the circulation. In addition, we report the novel finding that a subpopulation of endothelial cells in kidney, the main organ of polyomavirus persistence, showed selective and rapid uptake of VLPs, suggesting a role in viremic organ tropism.

Show MeSH
Related in: MedlinePlus