Epstein Barr virus (EBV) gains access to the host through tonsillar crypts. Our aim was to characterize microenvironment composition around EBV+ cells in tonsils from pediatric carriers, to disclose its role on viral pathogenesis. LMP1 expression, assessed by immunohistochemistry (IHC), was used to discriminate EBV + and – zones in 41 tonsil biopsies. Three regions were defined: Subepithelial (SE), interfollicular (IF) and germinal center (GC). CD8, GrB, CD68, IL10, Foxp3, PD1, CD56 and CD4 markers were evaluated by IHC; positive cells/100 total cells were counted. CD8+, GrB+, CD68+ and IL10+ cells were prevalent in EBV+ zones at the SE region (p < 0.0001, p = 0.03, p = 0.002 and p = 0.002 respectively, Wilcoxon test). CD4+ and CD68+ cell count were higher in EBV + GC (p = 0.01 and p = 0.0002 respectively, Wilcoxon test). Increment of CD8, GrB and CD68 at the SE region could indicate a specific response that may be due to local homing at viral entry, which could be counterbalanced by IL10, an immunosuppressive cytokine. Additionally, it could be hypothesized that CD4 augment at the GC may be involved in the EBV-induced B-cell growth control at this region, in which macrophages could also participate.
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This study was supported in part by a Grant from National Agency for Science and Technology Promotion (PICT 2014 nº0748 and PIDC 2013 n°048). P.Ch. and MVP are members of the National Research Council (CONICET), Research Career Program, MC is a CONICET postdoctoral fellow and AGV is a CONICET doctoral fellow. The authors thank Barbara Cao; Silvana Romero and Maria Jose Andrade (Histopathological Laboratory, at the Ricardo Gutierrez Children’s Hospital) for his helpful histotechnical work and the Otorhinolaryngology Service at the Ricardo Gutierrez Children’s Hospital who kindly help us and provide us with the samples.
Institutional guidelines regarding human experimentation were followed, according to the Helsinki Declaration of 1975. The protocol was approved by the Ethical Committee of our hospital, and written informed assent and consent was obtained from all patients or patient´s parents depending on age.
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The authors declare no conflict of interest.
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Supplementary Fig. 1: EBNA2 cytoplasmic positive staining at the germinal center macrophages, located in tonsils from pediatric EBV carriers. The stains were developed using diaminobenzidine (DAB). Digital images were obtained with an AxioCamErc 5s (Zeiss) camera and acquired using Digital Axio Vision Rel. 4.8 image acquisition software. Original magnification: 1000X (DOCX 10 KB)
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Vistarop, A.G., Cohen, M., Huaman, F. et al. The interplay between local immune response and Epstein–Barr virus-infected tonsillar cells could lead to viral infection control. Med Microbiol Immunol 207, 319–327 (2018). https://doi.org/10.1007/s00430-018-0553-2
- Epstein Barr virus
- Immune response