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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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Detection of EBER using in situ hybridization.(A-F) In situ detection of EBER-expressing cells in periapical granulomas (6 out of 9) was shown in the cytoplasm and nuclei of B cells (arrows) and plasma cells (arrow heads). (G-I) Three periapical granulomas showed EBER-negative expression. (J) Healthy gingival tissues never showed positive expression. Scale bar = 50μm.
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pone.0121548.g004: Detection of EBER using in situ hybridization.(A-F) In situ detection of EBER-expressing cells in periapical granulomas (6 out of 9) was shown in the cytoplasm and nuclei of B cells (arrows) and plasma cells (arrow heads). (G-I) Three periapical granulomas showed EBER-negative expression. (J) Healthy gingival tissues never showed positive expression. Scale bar = 50μm.

Mentions: In situ detection using EBER-specific probes was performed for periapical granulomas and healthy gingival tissues. In total, six of nine periapical granulomas (66.7%) showed positive expression of EBER (Fig 4A–4F). EBER localization was exhibited in the nucleus and cytoplasm of B lymphocytes and plasma cells. In contrast, EBV-negative periapical granulomas (Fig 4G–4I) and healthy gingival tissues (Fig 4J) did not show EBER positivity. No expression was detected using a sense probe for EBER (data not shown).


Epstein-Barr virus infection in chronically inflamed periapical granulomas.

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

Detection of EBER using in situ hybridization.(A-F) In situ detection of EBER-expressing cells in periapical granulomas (6 out of 9) was shown in the cytoplasm and nuclei of B cells (arrows) and plasma cells (arrow heads). (G-I) Three periapical granulomas showed EBER-negative expression. (J) Healthy gingival tissues never showed positive expression. Scale bar = 50μm.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0121548.g004: Detection of EBER using in situ hybridization.(A-F) In situ detection of EBER-expressing cells in periapical granulomas (6 out of 9) was shown in the cytoplasm and nuclei of B cells (arrows) and plasma cells (arrow heads). (G-I) Three periapical granulomas showed EBER-negative expression. (J) Healthy gingival tissues never showed positive expression. Scale bar = 50μm.
Mentions: In situ detection using EBER-specific probes was performed for periapical granulomas and healthy gingival tissues. In total, six of nine periapical granulomas (66.7%) showed positive expression of EBER (Fig 4A–4F). EBER localization was exhibited in the nucleus and cytoplasm of B lymphocytes and plasma cells. In contrast, EBV-negative periapical granulomas (Fig 4G–4I) and healthy gingival tissues (Fig 4J) did not show EBER positivity. No expression was detected using a sense probe for EBER (data not shown).

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