Characterization of Novel Splice Variants of Zinc Finger Antiviral Protein (ZAP).

Given the unprecedented scale of the recent Ebola and Zika viral epidemics, it is crucial to understand the biology of host factors with broad antiviral action in order to develop novel therapeutic approaches. Here, we look into one such factor: zinc finger antiviral protein (ZAP) inhibits a variety of RNA and DNA viruses. Alternative splicing results in two isoforms that differ at their C termini: ZAPL (long) encodes a poly(ADP-ribose) polymerase (PARP)-like domain that is missing in ZAPS (short). Previously, it has been shown that ZAPL is more antiviral than ZAPS, while the latter is more induced by interferon (IFN). In this study, we discovered and confirmed the expression of two additional splice variants of human ZAP: ZAPXL (extralong) and ZAPM (medium). We also found two haplotypes of human ZAP. Since ZAPL and ZAPS have differential activities, we hypothesize that all four ZAP isoforms have evolved to mediate distinct antiviral and/or cellular functions. By taking a gene-knockout-and-reconstitution approach, we have characterized the antiviral, translational inhibition, and IFN activation activities of individual ZAP isoforms. Our work demonstrates that ZAPL and ZAPXL are more active against alphaviruses and hepatitis B virus (HBV) than ZAPS and ZAPM and elucidates the effects of splice variants on the action of a broad-spectrum antiviral factor.IMPORTANCE ZAP is an IFN-induced host factor that can inhibit a wide range of viruses, and there is great interest in fully characterizing its antiviral mechanism. This is the first study that defines the antiviral capacities of individual ZAP isoforms in the absence of endogenous ZAP expression and, hence, cross talk with other isoforms. Our data demonstrate that ZAP is expressed as four different forms: ZAPS, ZAPM, ZAPL, and ZAPXL. The longer ZAP isoforms better inhibit alphaviruses and HBV, while all isoforms equally inhibit Ebola virus transcription and replication. In addition, there is no difference in the abilities of ZAP isoforms to enhance the induction of type I IFN expression. Our results show that the full spectrum of ZAP activities can change depending on the virus target and the relative levels of basal expression and induction by IFN or infection.

Cutting Edge: Drebrin-Regulated Actin Dynamics Regulate IgE-Dependent Mast Cell Activation and Allergic Responses.

Mast cells play critical roles in allergic responses. Calcium signaling controls the function of these cells, and a role for actin in regulating calcium influx into cells has been suggested. We have previously identified the actin reorganizing protein Drebrin as a target of the immunosuppressant 3,5-bistrifluoromethyl pyrazole, which inhibits calcium influx into cells. In this study, we show that Drebrin(-/-) mice exhibit reduced IgE-mediated histamine release and passive systemic anaphylaxis, and Drebrin(-/-) mast cells also exhibit defects in FcεRI-mediated degranulation. Drebrin(-/-) mast cells exhibit defects in actin cytoskeleton organization and calcium responses downstream of the FcεRI, and agents that relieve actin reorganization rescue mast cell FcεRI-induced degranulation. Our results indicate that Drebrin regulates the actin cytoskeleton and calcium responses in mast cells, thus regulating mast cell function in vivo.

Drebrin Regulation of Calcium Signaling in Immune Cells.

Store-operated Ca2+ channels are plasma membrane channels that are activated by depletion of intracellular Ca2+ stores, resulting in an increase in intracellular Ca2+; however, little is known about their regulation. Our work has shown that the immunosuppressant compound BTP2, which blocks Ca2+ influx into cells, interacts with the actin-reorganizing protein, drebrin. Here we review the role of drebrin in the regulation of calcium signaling, with a focus on immune cells.

Engineered bacteria launch and control an oncolytic virus.

The ability of bacteria and viruses to selectively replicate in tumors has led to synthetic engineering of new microbial therapies. Here we design a cooperative strategy whereby S. typhimurium bacteria transcribe and deliver the Senecavirus A RNA genome inside host cells, launching a potent oncolytic viral infection. Then, we engineer the virus to require a bacterially delivered protease in order to achieve virion maturation, demonstrating bacterial control over the virus. This work extends bacterially delivered therapeutics to viral genomes, and the governing of a viral population through engineered microbial interactions. One-Sentence Summary: Bacteria are engineered to act as a synthetic "capsid" delivering Senecavirus A genome and controlling its spread.

A robust cell culture system supporting the complete life cycle of hepatitis B virus.

The discovery of sodium taurocholate cotransporting polypeptide (NTCP) as the hepatitis B virus (HBV) receptor enabled researchers to create hepatoma cell lines susceptible to HBV infection. Infection in current systems, however, is inefficient and virus fails to spread. Infection efficiency is enhanced by treating cells with polyethylene glycol 8000 (PEG) during infection. However, this alone does not promote virus spread. Here we show that maintaining PEG in culture medium increases the rate of infection by at least one order of magnitude, and, most importantly, promotes virus spread. To demonstrate the utility of this system, we show that two interferon-stimulated genes (ISGs), ISG20 and tetherin, restrict HBV spread in NTCP-expressing hepatoma cells. Thus, this protocol can be easily applied to existing cell culture systems to study the complete HBV life cycle, including virus spread.

Spinal pleomorphic xanthoastrocytoma: case report and literature review / Xantoastrocitoma pleomórfico espinal: reporte de caso y revisión de la literatura

El Xantoastrocitoma Pleomórfico Espinal (XAPE) es una neoplasia primaria infrecuente del sistema nervioso central, descrita por primer vez en 1979. De estirpe astrocítica y aunque clasificada como grado II por la OMS, pueden existir variantes anaplásicas o malignas. Con localización usualmente temporal, pocos casos espinales se han descrito a la fecha, debido probablemente a un subdiagnóstico o un subregistro. El pronóstico es generalmente bueno y depende en gran medida del tipo histológico y extensión de la resección quirurgica. Describimos el caso de un hombre de 30 años de edad con dorsalgiay paresia del miembro inferior izquierdo. Se encontró en imágenes de resonancia magnética una lesión aparentemente intradural, extraaxial en los segmentos T8-T9 que se llevó a resección. Se confirmó por histopatología la presencia de un XAPE primario. En la presente publicación realizamos una revisión de la literatura disponible.

Spinal Pleomorphic Xanthoastrocytoma (SPXA) is a rare CNS primary neoplasm, first described in 1979. Although of astrocytic lineage and classified as a grade II neoplasm by the WHO, it may be have anaplastic or malignant variants. Usually located in the temporal lobe, few spinal cases have been described to date, probably due to underdiagnosis and underreporting. It usually has a good prognosis, but it depends on its histological type and extent of surgical resection. In this article, we describe the case of a 30-year old male who complained of low back pain and left lower limb paresis. The MRI showed an apparently intradural, extraaxial lesion at T8-T9 segments. The diagnosis of a primary SPXA was confirmed by histopathological studies. In this article, a review of the available literature is presented.