Limits...
Serological cross-reactivity between Merkel cell polyomavirus and two closely related chimpanzee polyomaviruses.

Nicol JT, Liais E, Potier R, Mazzoni E, Tognon M, Coursaget P, Touzé A - PLoS ONE (2014)

Bottom Line: Phylogenetic analyses based on the major capsid protein sequence indicate that Merkel cell polyomavirus (MCPyV) and chimpanzee polyomaviruses (PtvPyV1, PtvPyV2), and similarly Trichodysplasia spinulosa-associated polyomavirus (TSPyV) and the orangutan polyomavirus (OraPyV1) are closely related.In contrast, cross-reactivity was not observed between TSPyV and OraPyV1.Furthermore, specific antibodies to chimpanzee polyomaviruses were detected in chimpanzee sera by pre-incubation of sera with the different antigens, but not in human sera.

View Article: PubMed Central - PubMed

Affiliation: Université François Rabelais, Virologie Immunologie Moléculaires, Tours, France; INRA UMR 1282, Infectiologie et Santé Publique, Tours, France.

ABSTRACT
Phylogenetic analyses based on the major capsid protein sequence indicate that Merkel cell polyomavirus (MCPyV) and chimpanzee polyomaviruses (PtvPyV1, PtvPyV2), and similarly Trichodysplasia spinulosa-associated polyomavirus (TSPyV) and the orangutan polyomavirus (OraPyV1) are closely related. The existence of cross-reactivity between these polyomaviruses was therefore investigated. The findings indicated serological identity between the two chimpanzee polyomaviruses investigated and a high level of cross-reactivity with Merkel cell polyomavirus. In contrast, cross-reactivity was not observed between TSPyV and OraPyV1. Furthermore, specific antibodies to chimpanzee polyomaviruses were detected in chimpanzee sera by pre-incubation of sera with the different antigens, but not in human sera.

Show MeSH
Electron micrographs of VP1 virus-like particles for MCPyV, PtvPyV1b, PtvPyV2c, TSPyV and OraPyV1.VP1 were produced using recombinant baculoviruses. The preparations were applied to carbon-coated grids, negatively stained with 1.5% uranyl acetate and observed at 50,000 nominal magnification with a JEOL 1011 electron microscope (Scale bars, 100 nm).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4016208&req=5

pone-0097030-g002: Electron micrographs of VP1 virus-like particles for MCPyV, PtvPyV1b, PtvPyV2c, TSPyV and OraPyV1.VP1 were produced using recombinant baculoviruses. The preparations were applied to carbon-coated grids, negatively stained with 1.5% uranyl acetate and observed at 50,000 nominal magnification with a JEOL 1011 electron microscope (Scale bars, 100 nm).

Mentions: Production of MCPyV and TSPyV VLPs in insect cells has been described previously [15], [28]. VLPs were also generated for PtvPyV1, PtvPyV2 and OraPyV1. Briefly, VP1 coding sequences were obtained by total synthesis with codon usage-adapted sequences for expression in Spodoptera frugiperda cells (Genscript, Piscataway, NJ, USA) (Sequences based on PtvPyV1b (HQ385747), PtvPyV2c (HQ385750) and OraPyV1 (FN356900)) [26], [27]. The different VP1 genes were cloned under the control of the polyhedrin promoter of pFastBac Dual plasmid (Invitrogen, FisherScientific, Illkirch, France) and then used to generate recombinant baculoviruses using the Bac-to-Bac system (Invitrogen). HiFive cells maintained in Grace medium (Invitrogen) were infected with the different recombinant baculoviruses for production of the polyomavirus VLPs. VLPs were then purified by ultracentrifugation (18 h at 30,000 rpm in a Beckman SW 32 rotor) in a CsCl gradient and assembly of VP1 into VLPs was verified by electron microscopy. The preparations were applied to carbon grids, negatively stained with 1.5% uranyl acetate and observed with a JEOL 1011 electron microscope at 50,000 nominal magnification (Figure 2).


Serological cross-reactivity between Merkel cell polyomavirus and two closely related chimpanzee polyomaviruses.

Nicol JT, Liais E, Potier R, Mazzoni E, Tognon M, Coursaget P, Touzé A - PLoS ONE (2014)

Electron micrographs of VP1 virus-like particles for MCPyV, PtvPyV1b, PtvPyV2c, TSPyV and OraPyV1.VP1 were produced using recombinant baculoviruses. The preparations were applied to carbon-coated grids, negatively stained with 1.5% uranyl acetate and observed at 50,000 nominal magnification with a JEOL 1011 electron microscope (Scale bars, 100 nm).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4016208&req=5

pone-0097030-g002: Electron micrographs of VP1 virus-like particles for MCPyV, PtvPyV1b, PtvPyV2c, TSPyV and OraPyV1.VP1 were produced using recombinant baculoviruses. The preparations were applied to carbon-coated grids, negatively stained with 1.5% uranyl acetate and observed at 50,000 nominal magnification with a JEOL 1011 electron microscope (Scale bars, 100 nm).
Mentions: Production of MCPyV and TSPyV VLPs in insect cells has been described previously [15], [28]. VLPs were also generated for PtvPyV1, PtvPyV2 and OraPyV1. Briefly, VP1 coding sequences were obtained by total synthesis with codon usage-adapted sequences for expression in Spodoptera frugiperda cells (Genscript, Piscataway, NJ, USA) (Sequences based on PtvPyV1b (HQ385747), PtvPyV2c (HQ385750) and OraPyV1 (FN356900)) [26], [27]. The different VP1 genes were cloned under the control of the polyhedrin promoter of pFastBac Dual plasmid (Invitrogen, FisherScientific, Illkirch, France) and then used to generate recombinant baculoviruses using the Bac-to-Bac system (Invitrogen). HiFive cells maintained in Grace medium (Invitrogen) were infected with the different recombinant baculoviruses for production of the polyomavirus VLPs. VLPs were then purified by ultracentrifugation (18 h at 30,000 rpm in a Beckman SW 32 rotor) in a CsCl gradient and assembly of VP1 into VLPs was verified by electron microscopy. The preparations were applied to carbon grids, negatively stained with 1.5% uranyl acetate and observed with a JEOL 1011 electron microscope at 50,000 nominal magnification (Figure 2).

Bottom Line: Phylogenetic analyses based on the major capsid protein sequence indicate that Merkel cell polyomavirus (MCPyV) and chimpanzee polyomaviruses (PtvPyV1, PtvPyV2), and similarly Trichodysplasia spinulosa-associated polyomavirus (TSPyV) and the orangutan polyomavirus (OraPyV1) are closely related.In contrast, cross-reactivity was not observed between TSPyV and OraPyV1.Furthermore, specific antibodies to chimpanzee polyomaviruses were detected in chimpanzee sera by pre-incubation of sera with the different antigens, but not in human sera.

View Article: PubMed Central - PubMed

Affiliation: Université François Rabelais, Virologie Immunologie Moléculaires, Tours, France; INRA UMR 1282, Infectiologie et Santé Publique, Tours, France.

ABSTRACT
Phylogenetic analyses based on the major capsid protein sequence indicate that Merkel cell polyomavirus (MCPyV) and chimpanzee polyomaviruses (PtvPyV1, PtvPyV2), and similarly Trichodysplasia spinulosa-associated polyomavirus (TSPyV) and the orangutan polyomavirus (OraPyV1) are closely related. The existence of cross-reactivity between these polyomaviruses was therefore investigated. The findings indicated serological identity between the two chimpanzee polyomaviruses investigated and a high level of cross-reactivity with Merkel cell polyomavirus. In contrast, cross-reactivity was not observed between TSPyV and OraPyV1. Furthermore, specific antibodies to chimpanzee polyomaviruses were detected in chimpanzee sera by pre-incubation of sera with the different antigens, but not in human sera.

Show MeSH