J Korean Med Sci.  2017 Dec;32(12):1959-1966. 10.3346/jkms.2017.32.12.1959.

Identification of Epstein-Barr Virus in the Human Placenta and Its Pathologic Characteristics

• Kim Y
• Kim HS
• Park JS
• Kim CJ
• Kim WH

• KMID: 2396372
• DOI: http://doi.org/10.3346/jkms.2017.32.12.1959

Abstract

Epstein-Barr virus (EBV), a common pathogen in humans, is suspected as the cause of multiple pregnancy-related pathologies including depression, preeclampsia, and stillbirth. Moreover, transmission of EBV through the placenta has been reported. However, the focus of EBV infection within the placenta has remained unknown to date. In this study, we proved the expression of latent EBV genes in the endometrial glandular epithelial cells of the placenta and investigated the cytological characteristics of these cells. Sixty-eight placentas were obtained from pregnant women. Tissue microarray was constructed. EBV latent genes including EBV-encoding RNA-1 (EBER1), Epstein-Barr virus nuclear antigen 1 (EBNA1), late membrane antigen (LMP1), and RPMS1 were detected with silver in situ hybridization and/or mRNA in situ hybridization. Nuclear features of EBV-positive cells in EBV-infected placenta were compared with those of EBV-negative cells via image analysis. Sixteen placentas (23.5%) showed positive expression of all 4 EBV latent genes; only the glandular epithelial cells of the decidua showed EBV gene expression. EBV infection status was not significantly correlated with maternal, fetal, or placental factors. The nuclei of EBV-positive cells were significantly larger, longer, and round-shaped than those of EBV-negative cells regardless of EBV-infection status of the placenta. For the first time, evidence of EBV gene expression has been shown in placental tissues. Furthermore, we have characterized its cytological features, allowing screening of EBV infection through microscopic examination.

Keyword

Human Herpesvirus 4; Image Cytometry; In Situ Hybridization; Virus Latency; Decidua Capsularis

MeSH Terms

Decidua
Depression
Epithelial Cells
Epstein-Barr Virus Infections
Female
Gene Expression
Herpesvirus 4, Human*
Humans*
Image Cytometry
In Situ Hybridization
Mass Screening
Membranes
Pathology
Placenta*
Pre-Eclampsia
Pregnant Women
RNA, Messenger
Silver
Stillbirth
Virus Latency
RNA, Messenger
Silver

Figure

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  Fig. 1 Expression of various viral genes in an EBV-positive case. Hematoxylin-eosin staining (A). Positive staining is represented by black color in SISH of EBER1 gene by Ventana (B). ISH staining by RNAscopy of four EBV latent genes, including EBER1 (C), EBNA1 (D), LMP1 (E), and RPMS1 (F) was positive demonstrating the brown color at the nucleus. EBV = Epstein-Barr virus, SISH = silver in situ hybridization, EBER1 = EBV-encoding RNA-1, ISH = in situ hybridization, EBNA1 = Epstein-Barr virus nuclear antigen-1, LMP1 = late membrane antigen.

• 

  Fig. 2 Comparison of endometrial glands in the decidua of EBV-infected (left) and EBV-uninfected (right) chorioamniotic membranes. EBER1 SISH (A), EBER1 mRNA ISH (B), and EBNA1 mRNA ISH (C) were positive only in EBV-infected cases. Immunohistochemical staining of epithelial cells was positive for E-cadherin (D) and cytokeratin (E) and was negative for CD31 (F) regardless of EBV infection. EBV = Epstein-Barr virus, EBER1 = EBV-encoding RNA-1, SISH = silver in situ hybridization, ISH = in situ hybridization, EBNA1 = Epstein-Barr virus nuclear antigen-1.

• 

  Fig. 3 Cytological difference between EBV-positive (left) and EBV-negative glandular epithelial cells. Epithelial cells positive for EBV latent gene expression show drastic cellular changes including nuclear inclusion, less condense chromatin contour, increased nucleus and cytoplasm in volume, and perinuclear clearing of cytoplasm. EBV = Epstein-Barr virus.

• 

  Fig. 4 Example of difference in nuclear morphology based on EBV-positivity of individual cells. These nuclei were included in the analysis for objective comparison (Supplementary Table 2). EBV = Epstein-Barr virus.

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