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Epstein-Barr virus infection in chronically inflamed periapical granulomas.

Makino K, Takeichi O, Hatori K, Imai K, Ochiai K, Ogiso B - PLoS ONE (2015)

Bottom Line: In contrast, EBV DNA was not detected in healthy gingival tissues (n = 10); the difference was statistically significant according to the Mann-Whitney U test (p = 0.0001).Paraffin sections were also analyzed by in situ hybridization to detect EBV-encoded small RNA (EBER)-expressing cells.In addition, immunohistochemical analysis for latent membrane protein 1 (LMP-1) of EBV using serial tissue sections showed that LMP-1-expressing cells were localized to the same areas as EBER-expressing cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Endodontics, Nihon University School of Dentistry, Chiyoda-ku, Tokyo, Japan.

ABSTRACT
Periapical granulomas are lesions around the apex of a tooth caused by a polymicrobial infection. Treatment with antibacterial agents is normally performed to eliminate bacteria from root canals; however, loss of the supporting alveolar bone is typically observed, and tooth extraction is often selected if root canal treatment does not work well. Therefore, bacteria and other microorganisms could be involved in this disease. To understand the pathogenesis of periapical granulomas more precisely, we focused on the association with Epstein-Barr virus (EBV) using surgically removed periapical granulomas (n = 32). EBV DNA was detected in 25 of 32 periapical granulomas (78.1%) by real-time PCR, and the median number of EBV DNA copies was approximately 8,688.01/μg total DNA. In contrast, EBV DNA was not detected in healthy gingival tissues (n = 10); the difference was statistically significant according to the Mann-Whitney U test (p = 0.0001). Paraffin sections were also analyzed by in situ hybridization to detect EBV-encoded small RNA (EBER)-expressing cells. EBER was detected in the cytoplasm and nuclei of B cells and plasma cells in six of nine periapical granulomas, but not in healthy gingival tissues. In addition, immunohistochemical analysis for latent membrane protein 1 (LMP-1) of EBV using serial tissue sections showed that LMP-1-expressing cells were localized to the same areas as EBER-expressing cells. These data suggest that B cells and plasma cells in inflamed granulomas are a major source of EBV infection, and that EBV could play a pivotal role in controlling immune cell responses in periapical granulomas.

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Related in: MedlinePlus

Quantitative real time PCR analysis.(A) Standard curve to determine the copies of EBV DNA in specimens. EBV DNA (approximately 1x106 copies/μl) was diluted in ten-fold serially. Each diluted DNA was amplified using real-time PCR simultaneously at the time of amplification for DNA extracted from periapical granulomas and healthy gingivae. (B) Detection of EBV DNA in periapical granulomas and healthy gingivae. The copy of EBV DNA in each specimen was calculated using the standard curve. The median of EBV DNA copies in periapical granulomas was approximately 8688.01 per 1μg of total DNA, as shown by a horizontal bar. * showed statistical difference using Mann-Whitney U test (p = 0.0001).
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pone.0121548.g003: Quantitative real time PCR analysis.(A) Standard curve to determine the copies of EBV DNA in specimens. EBV DNA (approximately 1x106 copies/μl) was diluted in ten-fold serially. Each diluted DNA was amplified using real-time PCR simultaneously at the time of amplification for DNA extracted from periapical granulomas and healthy gingivae. (B) Detection of EBV DNA in periapical granulomas and healthy gingivae. The copy of EBV DNA in each specimen was calculated using the standard curve. The median of EBV DNA copies in periapical granulomas was approximately 8688.01 per 1μg of total DNA, as shown by a horizontal bar. * showed statistical difference using Mann-Whitney U test (p = 0.0001).

Mentions: DNA extracted from periapical granulomas and healthy gingival tissues was amplified using real-time PCR with EBV-specific primers. A standard curve was prepared after PCR amplification using serially diluted EBV DNA, as shown in Fig 3A, and PCR was performed at the same time as the amplification of DNA from periapical granulomas and healthy gingival tissues.


Epstein-Barr virus infection in chronically inflamed periapical granulomas.

Makino K, Takeichi O, Hatori K, Imai K, Ochiai K, Ogiso B - PLoS ONE (2015)

Quantitative real time PCR analysis.(A) Standard curve to determine the copies of EBV DNA in specimens. EBV DNA (approximately 1x106 copies/μl) was diluted in ten-fold serially. Each diluted DNA was amplified using real-time PCR simultaneously at the time of amplification for DNA extracted from periapical granulomas and healthy gingivae. (B) Detection of EBV DNA in periapical granulomas and healthy gingivae. The copy of EBV DNA in each specimen was calculated using the standard curve. The median of EBV DNA copies in periapical granulomas was approximately 8688.01 per 1μg of total DNA, as shown by a horizontal bar. * showed statistical difference using Mann-Whitney U test (p = 0.0001).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4401687&req=5

pone.0121548.g003: Quantitative real time PCR analysis.(A) Standard curve to determine the copies of EBV DNA in specimens. EBV DNA (approximately 1x106 copies/μl) was diluted in ten-fold serially. Each diluted DNA was amplified using real-time PCR simultaneously at the time of amplification for DNA extracted from periapical granulomas and healthy gingivae. (B) Detection of EBV DNA in periapical granulomas and healthy gingivae. The copy of EBV DNA in each specimen was calculated using the standard curve. The median of EBV DNA copies in periapical granulomas was approximately 8688.01 per 1μg of total DNA, as shown by a horizontal bar. * showed statistical difference using Mann-Whitney U test (p = 0.0001).
Mentions: DNA extracted from periapical granulomas and healthy gingival tissues was amplified using real-time PCR with EBV-specific primers. A standard curve was prepared after PCR amplification using serially diluted EBV DNA, as shown in Fig 3A, and PCR was performed at the same time as the amplification of DNA from periapical granulomas and healthy gingival tissues.

Bottom Line: In contrast, EBV DNA was not detected in healthy gingival tissues (n = 10); the difference was statistically significant according to the Mann-Whitney U test (p = 0.0001).Paraffin sections were also analyzed by in situ hybridization to detect EBV-encoded small RNA (EBER)-expressing cells.In addition, immunohistochemical analysis for latent membrane protein 1 (LMP-1) of EBV using serial tissue sections showed that LMP-1-expressing cells were localized to the same areas as EBER-expressing cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Endodontics, Nihon University School of Dentistry, Chiyoda-ku, Tokyo, Japan.

ABSTRACT
Periapical granulomas are lesions around the apex of a tooth caused by a polymicrobial infection. Treatment with antibacterial agents is normally performed to eliminate bacteria from root canals; however, loss of the supporting alveolar bone is typically observed, and tooth extraction is often selected if root canal treatment does not work well. Therefore, bacteria and other microorganisms could be involved in this disease. To understand the pathogenesis of periapical granulomas more precisely, we focused on the association with Epstein-Barr virus (EBV) using surgically removed periapical granulomas (n = 32). EBV DNA was detected in 25 of 32 periapical granulomas (78.1%) by real-time PCR, and the median number of EBV DNA copies was approximately 8,688.01/μg total DNA. In contrast, EBV DNA was not detected in healthy gingival tissues (n = 10); the difference was statistically significant according to the Mann-Whitney U test (p = 0.0001). Paraffin sections were also analyzed by in situ hybridization to detect EBV-encoded small RNA (EBER)-expressing cells. EBER was detected in the cytoplasm and nuclei of B cells and plasma cells in six of nine periapical granulomas, but not in healthy gingival tissues. In addition, immunohistochemical analysis for latent membrane protein 1 (LMP-1) of EBV using serial tissue sections showed that LMP-1-expressing cells were localized to the same areas as EBER-expressing cells. These data suggest that B cells and plasma cells in inflamed granulomas are a major source of EBV infection, and that EBV could play a pivotal role in controlling immune cell responses in periapical granulomas.

Show MeSH
Related in: MedlinePlus