Avilala, J. | bioRxiv Science

bioRxiv2026

Epstein-Barr Virus in HIV-Associated Conjunctival Squamous Cell Carcinoma and Transcriptional Reprogramming of Conjunctival Epithelial Cells

The involvement of Epstein-Barr virus (EBV) in conjunctival squamous cell carcinoma (cSCC) remains incompletely defined. This study integrated analyses of primary tumors with an experimental model of the conjunctival epithelium. Five Nigerian tumors from patients with HIV and three Japanese tumors from patients without HIV were examined by EBV-encoded RNA in situ hybridization (EBER-ISH), EBV DNA PCR, and virome analysis of RNA sequencing data. EBER localized to epithelial tumor cells in four Nigerian specimens and none of the Japanese specimens. EBV DNA and RNA were each detected in three Nigerian tumors, with every EBER-positive tumor supported by at least one additional assay. Human telomerase reverse transcriptase-immortalized conjunctival epithelial cells infected with Akata-GFP-EBV expressed latency-associated and lytic viral genes. Comparison of three infected and three control cultures identified 96 differentially expressed host genes and 23 enriched Hallmark gene sets at a false discovery rate below 0.05. RT-qPCR confirmed increased HES1, CXCL8, CXCL2, and LOX and decreased FGFBP1, CSTA, SAPCD2, and TMEM40 expression. Prominent responses involved inflammatory signaling, epithelial remodeling, metabolism, and cell-cycle regulation. These findings identify tumor-cell-associated EBV in a subset of HIV-associated cSCC and establish an epithelial model for investigating infection-associated host responses.

microbiology↗

bioRxiv2025

Interleukin-17 directly triggers Epstein-Barr virus lytic reactivation in latently infected human B cells

Epstein-Barr virus (EBV) is a ubiquitous human gammaherpesvirus implicated in a wide spectrum of inflammatory and malignant diseases. Although EBV toggles between latent and lytic states, the host cues that control this switch remain incompletely defined. Interleukin-17A (IL-17A; hereafter IL-17), a signature Th17 cytokine, is abundant in EBV-associated tissues, and EBV products can augment IL-17 responses. Whether IL-17 directly modulates the EBV life cycle has remained unknown. Here we show that IL-17 alone is sufficient to induce EBV lytic reactivation in latently infected human B cells. Across RT-qPCR, immunoblotting, and RNA-seq, IL-17 increased the immediate-early regulator BZLF1 (Zta) and upregulated downstream early and late viral genes, consistent with activation of a transcriptome-wide lytic program. Culture supernatants from IL-17-treated cells contained elevated DNase-resistant extracellular EBV DNA, indicating productive replication with encapsidated genomes. Transcriptomic pathway analyses confirmed engagement of IL-17-linked signaling and highlighted inflammatory modules, including NF-{kappa}B and JAK-STAT. Gene Ontology analysis further enriched for regulation of B-cell receptor signaling and B-cell activation, situating IL-17 within B-cell inflammatory circuitry. Together, these findings identify IL-17 as a cytokine cue for EBV lytic entry and provide a mechanistic link between Th17-skewed inflammation and episodic EBV reactivation. This cytokine-driven pathway complements existing models centered on B-cell receptor signaling, hypoxia/HIF-1, COX-2/PGE2, and TGF-{beta} and motivates testing whether modulation of the IL-17/IL-17R axis can alter EBV reactivation in IL-17-rich settings. To our knowledge, this is the first demonstration that IL-17 alone directly triggers EBV lytic reactivation in human cells. IMPORTANCEEpstein-Barr virus (EBV) persists for life by maintaining latency with intermittent lytic reactivation, yet the physiological cues that initiate the latency-lytic switch remain poorly defined. Here we identify the Th17 cytokine interleukin-17A (IL-17) as a direct host trigger of EBV lytic reactivation in latently infected human B cells. IL-17 alone induced the immediate-early transactivator Zta, activated a transcriptome-wide lytic program, and promoted release of DNase-resistant extracellular EBV DNA consistent with encapsidated virions. Transcriptomic analyses confirmed engagement of IL-17-linked inflammatory modules and implicated convergence on signaling nodes shared with canonical B-cell receptor pathways. These findings establish a mechanistic link between Th17-skewed inflammation and EBV reactivation burden, providing a framework to interrogate how IL-17-rich microenvironments influence EBV dissemination and immunopathology and to evaluate whether targeting the IL-17/IL-17R axis can modulate EBV reactivation in disease settings.

microbiology↗