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Binding and entry characteristics of porcine circovirus 2 in cells of the porcine monocytic line 3D4/31.
J Gen Virol. 2005 Jul; 86(Pt 7):2057-68.JG

Abstract

Porcine circovirus 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome and reproductive problems in pigs. Cells of the monocyte/macrophage lineage are important target cells in PCV2-infected pigs, but the method of binding and entry of PCV2 into these cells is unknown. Therefore, binding and entry of PCV2 to the porcine monocytic cell line 3D4/31 were studied by visualization of binding and internalization of PCV2 virus-like particles (VLPs) by confocal microscopy and chemical inhibition of endocytic pathways (clathrin- and caveolae-mediated endocytosis and macropinocytosis), followed by evaluation of the level of PCV2 infection. It was shown that PCV2 VLPs bound to all cells, with maximal binding starting from 30 min post-incubation. Bound PCV2 VLPs were internalized in 47+/-5.0 % of cells. Internalization was continuous, with 70.5+/-9.7 % of bound PCV2 VLPs internalized at 360 min post-incubation. Internalizing PCV2 VLPs co-localized with clathrin. PCV2 infection was decreased significantly by chemical inhibitors that specifically blocked (i) actin-dependent processes, including cytochalasin D (75.5+/-7.0 % reduction) and latrunculin B (71.0+/-3.0 % reduction), and (ii) clathrin-mediated endocytosis, including potassium depletion combined with hypotonic shock (50.2+/-6.3 % reduction), hypertonic medium (56.4+/-5.7 % reduction), cytosol acidification (59.1+/-7.1 % reduction) and amantadine (52.6+/-6.7 % reduction). Inhibiting macropinocytosis with amiloride and caveolae-dependent endocytosis with nystatin did not decrease PCV2 infection significantly. PCV2 infection was reduced by the lysosomotropic weak bases ammonium chloride (47.0+/-7.9 % reduction) and chloroquine diphosphate (49.0+/-5.6 % reduction). Together, these data demonstrate that PCV2 enters 3D4/31 cells predominantly via clathrin-mediated endocytosis and requires an acidic environment for infection.

Authors+Show Affiliations

Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15958685

Citation

Misinzo, G, et al. "Binding and Entry Characteristics of Porcine Circovirus 2 in Cells of the Porcine Monocytic Line 3D4/31." The Journal of General Virology, vol. 86, no. Pt 7, 2005, pp. 2057-68.
Misinzo G, Meerts P, Bublot M, et al. Binding and entry characteristics of porcine circovirus 2 in cells of the porcine monocytic line 3D4/31. J Gen Virol. 2005;86(Pt 7):2057-68.
Misinzo, G., Meerts, P., Bublot, M., Mast, J., Weingartl, H. M., & Nauwynck, H. J. (2005). Binding and entry characteristics of porcine circovirus 2 in cells of the porcine monocytic line 3D4/31. The Journal of General Virology, 86(Pt 7), 2057-68.
Misinzo G, et al. Binding and Entry Characteristics of Porcine Circovirus 2 in Cells of the Porcine Monocytic Line 3D4/31. J Gen Virol. 2005;86(Pt 7):2057-68. PubMed PMID: 15958685.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Binding and entry characteristics of porcine circovirus 2 in cells of the porcine monocytic line 3D4/31. AU - Misinzo,G, AU - Meerts,P, AU - Bublot,M, AU - Mast,J, AU - Weingartl,H M, AU - Nauwynck,H J, PY - 2005/6/17/pubmed PY - 2005/7/29/medline PY - 2005/6/17/entrez SP - 2057 EP - 68 JF - The Journal of general virology JO - J. Gen. Virol. VL - 86 IS - Pt 7 N2 - Porcine circovirus 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome and reproductive problems in pigs. Cells of the monocyte/macrophage lineage are important target cells in PCV2-infected pigs, but the method of binding and entry of PCV2 into these cells is unknown. Therefore, binding and entry of PCV2 to the porcine monocytic cell line 3D4/31 were studied by visualization of binding and internalization of PCV2 virus-like particles (VLPs) by confocal microscopy and chemical inhibition of endocytic pathways (clathrin- and caveolae-mediated endocytosis and macropinocytosis), followed by evaluation of the level of PCV2 infection. It was shown that PCV2 VLPs bound to all cells, with maximal binding starting from 30 min post-incubation. Bound PCV2 VLPs were internalized in 47+/-5.0 % of cells. Internalization was continuous, with 70.5+/-9.7 % of bound PCV2 VLPs internalized at 360 min post-incubation. Internalizing PCV2 VLPs co-localized with clathrin. PCV2 infection was decreased significantly by chemical inhibitors that specifically blocked (i) actin-dependent processes, including cytochalasin D (75.5+/-7.0 % reduction) and latrunculin B (71.0+/-3.0 % reduction), and (ii) clathrin-mediated endocytosis, including potassium depletion combined with hypotonic shock (50.2+/-6.3 % reduction), hypertonic medium (56.4+/-5.7 % reduction), cytosol acidification (59.1+/-7.1 % reduction) and amantadine (52.6+/-6.7 % reduction). Inhibiting macropinocytosis with amiloride and caveolae-dependent endocytosis with nystatin did not decrease PCV2 infection significantly. PCV2 infection was reduced by the lysosomotropic weak bases ammonium chloride (47.0+/-7.9 % reduction) and chloroquine diphosphate (49.0+/-5.6 % reduction). Together, these data demonstrate that PCV2 enters 3D4/31 cells predominantly via clathrin-mediated endocytosis and requires an acidic environment for infection. SN - 0022-1317 UR - https://www.unboundmedicine.com/medline/citation/15958685/Binding_and_entry_characteristics_of_porcine_circovirus_2_in_cells_of_the_porcine_monocytic_line_3D4/31_ L2 - http://jgv.microbiologyresearch.org/pubmed/content/journal/jgv/10.1099/vir.0.80652-0 DB - PRIME DP - Unbound Medicine ER -