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Productive infection of bovine papillomavirus type 2 in the urothelial cells of naturally occurring urinary bladder tumors in cattle and water buffaloes.

Roperto S, Russo V, Ozkul A, Corteggio A, Sepici-Dincel A, Catoi C, Esposito I, Riccardi MG, Urraro C, Lucà R, Ceccarelli DM, Longo M, Roperto F - PLoS ONE (2013)

Bottom Line: L1 DNA was amplified by PCR, cloned and sequenced and confirmed to be L1 DNA.Immunohistochemical investigations confirmed the presence of L1 protein both in the cytoplasm and nuclei of cells of the neoplastic urothelium.Electron microscopic investigations detected electron dense particles, the shape and size of which are consistent with submicroscopic features of viral particles, in nuclei of neoplastic urothelium.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Medicina Veterinaria e Produzioni Animali, Sezione Malattie Infettive, Università di Napoli Federico II, Naples, Italy. sante.roperto@unina.it

ABSTRACT

Background: Papillomaviruses (PVs) are highly epitheliotropic as they usually establish productive infections within squamous epithelia of the skin, the anogenital tract and the oral cavity. In this study, early (E) and late (L) protein expression of bovine papillomavirus type 2 (BPV-2) in the urothelium of the urinary bladder is described in cows and water buffaloes suffering from naturally occurring papillomavirus-associated urothelial bladder tumors.

Methods and findings: E5 protein, the major oncoprotein of the BPV-2, was detected in all tumors. L1 DNA was amplified by PCR, cloned and sequenced and confirmed to be L1 DNA. The major capsid protein, L1, believed to be only expressed in productive papillomavirus infection was detected by Western blot analysis. Immunohistochemical investigations confirmed the presence of L1 protein both in the cytoplasm and nuclei of cells of the neoplastic urothelium. Finally, the early protein E2, required for viral DNA replication and known to be a pivotal factor for both productive and persistent infection, was detected by Western blot and immunohistochemically. Electron microscopic investigations detected electron dense particles, the shape and size of which are consistent with submicroscopic features of viral particles, in nuclei of neoplastic urothelium.

Conclusion: This study shows that both active and productive infections by BPV-2 in the urothelium of the bovine and bubaline urinary bladder can occur in vivo.

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Related in: MedlinePlus

E2 expression.The presence of E2 protein detected by Western blot. a) Lanes 1–2: urinary bladders from healthy cows. Lanes 3–7: urinary bladder tumors from five of twenty cows. Lane 8: positive control, (bovine placenta infected with BPV-2). b) Lanes 1–2: urinary bladders from healthy buffaloes. Lanes 3–7: urinary bladder tumors from five of twenty-one buffaloes. Lane 8: positive control, (bovine placenta infected with BPV-2).
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pone-0062227-g005: E2 expression.The presence of E2 protein detected by Western blot. a) Lanes 1–2: urinary bladders from healthy cows. Lanes 3–7: urinary bladder tumors from five of twenty cows. Lane 8: positive control, (bovine placenta infected with BPV-2). b) Lanes 1–2: urinary bladders from healthy buffaloes. Lanes 3–7: urinary bladder tumors from five of twenty-one buffaloes. Lane 8: positive control, (bovine placenta infected with BPV-2).

Mentions: Since it is known that E2 protein is essential for the viral life cycle and plays a part in productive infection, molecular and immunohistochemical studies on the expression of this protein were also carried out. E2 protein expression was detected by Western blot analysis in total protein extracts from both bovine and bubaline urothelial tumors (Figure 5). E2 protein expression was also shown with morphological procedures. Immunohistochemical studies confirmed the presence of E2 protein in the cytoplasm and nuclei of neoplastic urothelial cells (Figure 6A). E2 protein expression was not detected in normal urothelial cells (Figure 6B).


Productive infection of bovine papillomavirus type 2 in the urothelial cells of naturally occurring urinary bladder tumors in cattle and water buffaloes.

Roperto S, Russo V, Ozkul A, Corteggio A, Sepici-Dincel A, Catoi C, Esposito I, Riccardi MG, Urraro C, Lucà R, Ceccarelli DM, Longo M, Roperto F - PLoS ONE (2013)

E2 expression.The presence of E2 protein detected by Western blot. a) Lanes 1–2: urinary bladders from healthy cows. Lanes 3–7: urinary bladder tumors from five of twenty cows. Lane 8: positive control, (bovine placenta infected with BPV-2). b) Lanes 1–2: urinary bladders from healthy buffaloes. Lanes 3–7: urinary bladder tumors from five of twenty-one buffaloes. Lane 8: positive control, (bovine placenta infected with BPV-2).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3646877&req=5

pone-0062227-g005: E2 expression.The presence of E2 protein detected by Western blot. a) Lanes 1–2: urinary bladders from healthy cows. Lanes 3–7: urinary bladder tumors from five of twenty cows. Lane 8: positive control, (bovine placenta infected with BPV-2). b) Lanes 1–2: urinary bladders from healthy buffaloes. Lanes 3–7: urinary bladder tumors from five of twenty-one buffaloes. Lane 8: positive control, (bovine placenta infected with BPV-2).
Mentions: Since it is known that E2 protein is essential for the viral life cycle and plays a part in productive infection, molecular and immunohistochemical studies on the expression of this protein were also carried out. E2 protein expression was detected by Western blot analysis in total protein extracts from both bovine and bubaline urothelial tumors (Figure 5). E2 protein expression was also shown with morphological procedures. Immunohistochemical studies confirmed the presence of E2 protein in the cytoplasm and nuclei of neoplastic urothelial cells (Figure 6A). E2 protein expression was not detected in normal urothelial cells (Figure 6B).

Bottom Line: L1 DNA was amplified by PCR, cloned and sequenced and confirmed to be L1 DNA.Immunohistochemical investigations confirmed the presence of L1 protein both in the cytoplasm and nuclei of cells of the neoplastic urothelium.Electron microscopic investigations detected electron dense particles, the shape and size of which are consistent with submicroscopic features of viral particles, in nuclei of neoplastic urothelium.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Medicina Veterinaria e Produzioni Animali, Sezione Malattie Infettive, Università di Napoli Federico II, Naples, Italy. sante.roperto@unina.it

ABSTRACT

Background: Papillomaviruses (PVs) are highly epitheliotropic as they usually establish productive infections within squamous epithelia of the skin, the anogenital tract and the oral cavity. In this study, early (E) and late (L) protein expression of bovine papillomavirus type 2 (BPV-2) in the urothelium of the urinary bladder is described in cows and water buffaloes suffering from naturally occurring papillomavirus-associated urothelial bladder tumors.

Methods and findings: E5 protein, the major oncoprotein of the BPV-2, was detected in all tumors. L1 DNA was amplified by PCR, cloned and sequenced and confirmed to be L1 DNA. The major capsid protein, L1, believed to be only expressed in productive papillomavirus infection was detected by Western blot analysis. Immunohistochemical investigations confirmed the presence of L1 protein both in the cytoplasm and nuclei of cells of the neoplastic urothelium. Finally, the early protein E2, required for viral DNA replication and known to be a pivotal factor for both productive and persistent infection, was detected by Western blot and immunohistochemically. Electron microscopic investigations detected electron dense particles, the shape and size of which are consistent with submicroscopic features of viral particles, in nuclei of neoplastic urothelium.

Conclusion: This study shows that both active and productive infections by BPV-2 in the urothelium of the bovine and bubaline urinary bladder can occur in vivo.

Show MeSH
Related in: MedlinePlus