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Epstein-Barr virus stimulates torque teno virus replication: a possible relationship to multiple sclerosis.

Borkosky SS, Whitley C, Kopp-Schneider A, zur Hausen H, de Villiers EM - PLoS ONE (2012)

Bottom Line: Our results demonstrate the replication of both transfected TTV genomes up to day 21 post transfection in all the evaluated cell lines.Quantitative amplification indicates statistically significant enhanced TTV replication in the EBV-positive cell lines, including the EBV-converted BJAB line, in comparison to the EBV-negative Burkitt's lymphoma cell line BJAB.This suggests a helper effect of EBV infections in the replication of TTV.

View Article: PubMed Central - PubMed

Affiliation: Division for the Characterization of Tumorviruses, Deutsches Krebsforschungszentrum, Heidelberg, Germany.

ABSTRACT
Viral infections have been implicated in the pathogenesis of multiple sclerosis. Epstein-Barr virus (EBV) has frequently been investigated as a possible candidate and torque teno virus (TTV) has also been discussed in this context. Nevertheless, mechanistic aspects remain unresolved. We report viral replication, as measured by genome amplification, as well as quantitative PCR of two TTV-HD14 isolates isolated from multiple sclerosis brain in a series of EBV-positive and -negative lymphoblastoid and Burkitt's lymphoma cell lines. Our results demonstrate the replication of both transfected TTV genomes up to day 21 post transfection in all the evaluated cell lines. Quantitative amplification indicates statistically significant enhanced TTV replication in the EBV-positive cell lines, including the EBV-converted BJAB line, in comparison to the EBV-negative Burkitt's lymphoma cell line BJAB. This suggests a helper effect of EBV infections in the replication of TTV. The present study provides information on a possible interaction of EBV and TTV in the etiology and progression of multiple sclerosis.

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In vitro replication of TTV-HD14b and TTV-HD14c as measured by long-PCR amplification.(A) PCR amplification of full-length TTV-HD14b in P3HR-1, BJAB, BJAB/EBV and full-length TTV-HD14c in ND1 and Ramos/EBV cell lines. Days after transfection are indicated. M - DNA size marker, C1 - untransfected cells, C2 - untransfected cells with nucleofector solution, + - positive control for long PCR amplification consisted either of re-ligated TTV-HD14b or TTV-HD14c mixed with Ramos cell line DNA. (B) Electron micrograph of B95-8 cells 3 days after transfection with linearized TTV-HD14b DNA. TTV-like particles can be seen within the nucleus of a cell. EBV particles are indicated by short arrows. Bar = 250 nm.
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pone-0032160-g001: In vitro replication of TTV-HD14b and TTV-HD14c as measured by long-PCR amplification.(A) PCR amplification of full-length TTV-HD14b in P3HR-1, BJAB, BJAB/EBV and full-length TTV-HD14c in ND1 and Ramos/EBV cell lines. Days after transfection are indicated. M - DNA size marker, C1 - untransfected cells, C2 - untransfected cells with nucleofector solution, + - positive control for long PCR amplification consisted either of re-ligated TTV-HD14b or TTV-HD14c mixed with Ramos cell line DNA. (B) Electron micrograph of B95-8 cells 3 days after transfection with linearized TTV-HD14b DNA. TTV-like particles can be seen within the nucleus of a cell. EBV particles are indicated by short arrows. Bar = 250 nm.

Mentions: We subsequently demonstrated the helper-dependent long-term replication and propagation of 12 TTV isolates in the 293TT cell line harbouring SV40 large T-antigen [67]. In the present study we analysed whether EBV is also able to exert a helper function for the replication of TTV. We included Burkitt's lymphoma cell lines both positive (P3HR-1, BJAB/EBV, Ramos/EBV) and negative for EBV (BJAB, Ramos), an EBV-immortalized B cell line (ND1) from an MS patient and an EBV-producing B cell line (B95-8). Replication of TTV-HD DNA was confirmed in all tested cell lines by long distance PCR on total cellular DNA. Long distance PCR was performed using back-to-back primers specific for amplifying the full-length genomes of TTV-HD14b and TTV-HD14c. Examples are presented in Figure 1A. Results were comparable between TTV-HD14b and TTV-HD14c and replication was measured up to day 21 after transfection in all cell lines. Replication was also confirmed by electron microscopy (Figure 1B).


Epstein-Barr virus stimulates torque teno virus replication: a possible relationship to multiple sclerosis.

Borkosky SS, Whitley C, Kopp-Schneider A, zur Hausen H, de Villiers EM - PLoS ONE (2012)

In vitro replication of TTV-HD14b and TTV-HD14c as measured by long-PCR amplification.(A) PCR amplification of full-length TTV-HD14b in P3HR-1, BJAB, BJAB/EBV and full-length TTV-HD14c in ND1 and Ramos/EBV cell lines. Days after transfection are indicated. M - DNA size marker, C1 - untransfected cells, C2 - untransfected cells with nucleofector solution, + - positive control for long PCR amplification consisted either of re-ligated TTV-HD14b or TTV-HD14c mixed with Ramos cell line DNA. (B) Electron micrograph of B95-8 cells 3 days after transfection with linearized TTV-HD14b DNA. TTV-like particles can be seen within the nucleus of a cell. EBV particles are indicated by short arrows. Bar = 250 nm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3285200&req=5

pone-0032160-g001: In vitro replication of TTV-HD14b and TTV-HD14c as measured by long-PCR amplification.(A) PCR amplification of full-length TTV-HD14b in P3HR-1, BJAB, BJAB/EBV and full-length TTV-HD14c in ND1 and Ramos/EBV cell lines. Days after transfection are indicated. M - DNA size marker, C1 - untransfected cells, C2 - untransfected cells with nucleofector solution, + - positive control for long PCR amplification consisted either of re-ligated TTV-HD14b or TTV-HD14c mixed with Ramos cell line DNA. (B) Electron micrograph of B95-8 cells 3 days after transfection with linearized TTV-HD14b DNA. TTV-like particles can be seen within the nucleus of a cell. EBV particles are indicated by short arrows. Bar = 250 nm.
Mentions: We subsequently demonstrated the helper-dependent long-term replication and propagation of 12 TTV isolates in the 293TT cell line harbouring SV40 large T-antigen [67]. In the present study we analysed whether EBV is also able to exert a helper function for the replication of TTV. We included Burkitt's lymphoma cell lines both positive (P3HR-1, BJAB/EBV, Ramos/EBV) and negative for EBV (BJAB, Ramos), an EBV-immortalized B cell line (ND1) from an MS patient and an EBV-producing B cell line (B95-8). Replication of TTV-HD DNA was confirmed in all tested cell lines by long distance PCR on total cellular DNA. Long distance PCR was performed using back-to-back primers specific for amplifying the full-length genomes of TTV-HD14b and TTV-HD14c. Examples are presented in Figure 1A. Results were comparable between TTV-HD14b and TTV-HD14c and replication was measured up to day 21 after transfection in all cell lines. Replication was also confirmed by electron microscopy (Figure 1B).

Bottom Line: Our results demonstrate the replication of both transfected TTV genomes up to day 21 post transfection in all the evaluated cell lines.Quantitative amplification indicates statistically significant enhanced TTV replication in the EBV-positive cell lines, including the EBV-converted BJAB line, in comparison to the EBV-negative Burkitt's lymphoma cell line BJAB.This suggests a helper effect of EBV infections in the replication of TTV.

View Article: PubMed Central - PubMed

Affiliation: Division for the Characterization of Tumorviruses, Deutsches Krebsforschungszentrum, Heidelberg, Germany.

ABSTRACT
Viral infections have been implicated in the pathogenesis of multiple sclerosis. Epstein-Barr virus (EBV) has frequently been investigated as a possible candidate and torque teno virus (TTV) has also been discussed in this context. Nevertheless, mechanistic aspects remain unresolved. We report viral replication, as measured by genome amplification, as well as quantitative PCR of two TTV-HD14 isolates isolated from multiple sclerosis brain in a series of EBV-positive and -negative lymphoblastoid and Burkitt's lymphoma cell lines. Our results demonstrate the replication of both transfected TTV genomes up to day 21 post transfection in all the evaluated cell lines. Quantitative amplification indicates statistically significant enhanced TTV replication in the EBV-positive cell lines, including the EBV-converted BJAB line, in comparison to the EBV-negative Burkitt's lymphoma cell line BJAB. This suggests a helper effect of EBV infections in the replication of TTV. The present study provides information on a possible interaction of EBV and TTV in the etiology and progression of multiple sclerosis.

Show MeSH
Related in: MedlinePlus