Child Protective Services Reports in the Context of Intimate Partner Violence: A Delphi Process Examining Best Practices.

OBJECTIVES: Child protective services (CPS) reporting for families experiencing intimate partner violence (IPV) is complex. The goal of this study was to develop expert-driven best practices for pediatric providers filing CPS reports in the context of IPV. METHODS: We conducted a Delphi study with experts in IPV and child abuse and neglect (CAN) through 3 rounds of surveys. In Round 1, participants selected clinical scenarios for which they would file, as well as best practices when CPS reporting is indicating. In Round 2, participants described how strongly they agreed that a provider should file for each clinical scenario and how important each best practice was on a 5-point Likert scale. Finally, in Round 3 participants reviewed Round 1 and 2 results, then reported their final determination by selecting yes or no for each option. Consensus was achieved in Round 3 if >80% of participants agreed. In each round, participants could provide further detail via free-text answers. RESULTS: Twenty-three (40%) of the invited experts participated. Consensus was not achieved for children directly witnessing IPV or experiencing health symptoms due to IPV exposure. Participants were in consensus regarding need for CPS reporting when CAN was present and that reporting should not occur for exposure to IPV only. Best practices included supporting IPV survivors, developing healthcare-based IPV advocacy programs, and optimizing the child welfare system. CONCLUSION: This study provides expert-driven recommendations for filing CPS reports in the context of IPV and highlights the inherent complexity of filing and the need for further guidelines.

Epstein-Barr virus microRNAs regulate B cell receptor signal transduction and lytic reactivation.

MicroRNAs (miRNAs) are post-transcriptional regulatory RNAs that can modulate cell signaling and play key roles in cell state transitions. Epstein-Barr virus (EBV) expresses >40 viral miRNAs that manipulate both viral and cellular gene expression patterns and contribute to reprogramming of the host environment during infection. Here, we identified a subset of EBV miRNAs that desensitize cells to B cell receptor (BCR) stimuli, and attenuate the downstream activation of NF-kappaB or AP1-dependent transcription. Bioinformatics and pathway analysis of Ago PAR-CLIP datasets identified multiple EBV miRNA targets related to BCR signal transduction, including GRB2, SOS1, MALT1, RAC1, and INPP5D, which we validated in reporter assays. BCR signaling is critical for B cell activation, proliferation, and differentiation, and for EBV, is linked to reactivation. In functional assays, we demonstrate that EBV miR-BHRF1-2-5p contributes to the growth of latently infected B cells through GRB2 regulation. We further determined that activities of EBV miR-BHRF1-2-5p, EBV miR-BART2-5p, and a cellular miRNA, miR-17-5p, directly regulate virus reactivation triggered by BCR engagement. Our findings provide mechanistic insight into some of the key miRNA interactions impacting the proliferation of latently infected B cells and importantly, governing the latent to lytic switch.

An Epstein-Barr Virus MicroRNA Blocks Interleukin-1 (IL-1) Signaling by Targeting IL-1 Receptor 1.

Epstein-Barr virus (EBV) encodes >44 viral microRNAs (miRNAs) that are differentially expressed throughout infection, can be detected in Epstein-Barr virus (EBV)-positive tumors, and manipulate several biological processes, including cell proliferation, apoptosis, and immune responses. Here, we show that EBV BHRF1-2 miRNAs block NF-κB activation following treatment with proinflammatory cytokines, specifically interleukin-1ß (IL-1ß). Analysis of EBV PAR-CLIP miRNA targetome data sets combined with pathway analysis revealed multiple BHRF1-2 miRNA targets involved in interleukin signaling pathways. By further analyzing changes in cellular gene expression patterns, we identified the IL-1 receptor 1 (IL1R1) as a direct target of miR-BHRF1-2-5p. Targeting the IL1R1 3' untranslated region (UTR) by EBV miR-BHRF1-2-5p was confirmed using 3'-UTR luciferase reporter assays and Western blot assays. Manipulation of EBV BHRF1-2 miRNA activity in latently infected B cells altered steady-state cytokine levels and disrupted IL-1ß responsiveness. These studies demonstrate functionally relevant BHRF1-2 miRNA interactions during EBV infection, which is an important step in understanding their roles in pathogenesis.IMPORTANCE IL-1 signaling plays an important role in inflammation and early activation of host innate immune responses following virus infection. Here, we demonstrate that a viral miRNA downregulates the IL-1 receptor 1 during EBV infection, which consequently alters the responsiveness of cells to IL-1 stimuli and changes the cytokine expression levels within infected cell populations. We postulate that this viral miRNA activity not only disrupts IL-1 autocrine and paracrine signaling loops that can alert effector cells to sites of infection but also provides a survival advantage by dampening excessive inflammation that may be detrimental to the infected cell.