Although mosquitoes are major transmission vectors for pathogenic arboviruses, viral infection has little impact on mosquito health. This immunity is caused in part by mosquito RNA interference (RNAi) pathways that generate antiviral small interfering RNAs (siRNAs) and Piwi-interacting RNAs (piRNAs). RNAi also maintains genome integrity by potently repressing mosquito transposon activity in the germline and soma. However, viral and transposon small RNA regulatory pathways have not been systematically examined together in mosquitoes. Therefore, we developed an integrated mosquito small RNA genomics (MSRG) resource that analyzes the transposon and virus small RNA profiles in mosquito cell cultures and somatic and gonadal tissues across four medically important mosquito species. Our resource captures both somatic and gonadal small RNA expression profiles within mosquito cell cultures, and we report the evolutionary dynamics of a novel Mosquito-Conserved piRNA Cluster Locus (MCpiRCL) made up of satellite DNA repeats. In the larger culicine mosquito genomes we detected highly regular periodicity in piRNA biogenesis patterns coinciding with the expansion of Piwi pathway genes. Finally, our resource enables detection of cross talk between piRNA and siRNA populations in mosquito cells during a response to virus infection. The MSRG resource will aid efforts to dissect and combat the capacity of mosquitoes to tolerate and spread arboviruses.

1Although mosquitoes are major transmission vectors for pathogenic arboviruses, 2 viral infection has little impact on mosquito health. This immunity is due in part to 3 mosquito RNA interference (RNAi) pathways that generate antiviral small interfering 4 RNAs (siRNAs) and Piwi-interacting RNAs (piRNAs). RNAi also maintains genome 5 integrity by potently repressing mosquito transposon activity in the germline and soma. 6 However, viral and transposon small RNA regulatory pathways have not been 7 systematically examined together in mosquitoes. Therefore, we developed an integrated 8 Mosquito Small RNA Genomics (MSRG) resource that analyzes the transposon and 9 virus small RNA profiles in mosquito cell cultures and somatic and gonadal tissues 10 across four medically important mosquito species. Our resource captures both somatic 11 and gonadal small RNA expression profiles within mosquito cell cultures, and we report 12 the evolutionary dynamics of a novel Mosquito-Conserved piRNA Cluster Locus 13 (MCpiRCL) composed of satellite DNA repeats. In the larger culicine mosquito genomes 14 we detected highly regular periodicity in piRNA biogenesis patterns coinciding with the 15

Common variable immunodeficiency (CVID) is the most frequently diagnosed primary antibody deficiency. About half of CVID patients develop chronic non-infectious complications thought to be due to intrinsic immune dysregulation, including autoimmunity, gastrointestinal disease, and interstitial lung disease (ILD). Multiple studies have found ILD to be a significant cause of morbidity and mortality in CVID. Yet, the precise mechanisms underlying this complication in CVID are poorly understood. CVID ILD is marked by profound pulmonary infiltration of both T and B cells as well as granulomatous inflammation in many cases. B cell depletive therapy, whether done as a monotherapy or in combination with another immunosuppressive agent, has become a standard of therapy for CVID ILD. However, CVID is a heterogeneous disorder, as is its lung pathology, and the precise patients that would benefit from B cell depletive therapy, when it should administered, and how long it should be repeated all remain gaps in our knowledge. Moreover, some have ILD recurrence after B cell depletive therapy and the relative importance of B cell biology remains incompletely defined. Developmental and functional abnormalities of B cell compartments observed in CVID ILD and related conditions suggest that imbalance of B cell signaling networks may promote lung disease. Included within these potential mechanisms of disease is B cell activating factor (BAFF), a cytokine that is upregulated by the interferon gamma (IFN-γ):STAT1 signaling axis to potently influence B cell activation and survival. B cell responses to BAFF are shaped by the divergent effects and expression patterns of its three receptors: BAFF receptor (BAFF-R), transmembrane activator and CAML interactor (TACI), and B cell maturation antigen (BCMA). Moreover, soluble forms of BAFF-R, TACI, and BCMA exist and may further influence the pathogenesis of ILD. Continued efforts to understand how dysregulated B cell biology promotes ILD development and progression will help close the gap in our understanding of how to best diagnose, define, and manage ILD in CVID.

Miceand other small rodent animal models are known to have greater resistance tointoxication by the organophosphorus (OP) nerve agents sarin, soman, andcyclosarin than do humans and non‐human primates. This resistance has beendirectly attributed to the presence of carboxylesterase in the blood plasma of these animals. Carboxylesterase acts asan endogenous bioscavenger and is not found in the blood plasma of humans andnon‐human primates. To create an improved small animal model of nerve agentintoxication, the gene encoding serum carboxylesterase (Es1) in C57BL/6 mice was deleted to generate a transgenic strain of mice (Es1 KO) that no longer expresses this protein. In contrast with previousgenetic modification efforts to remove other endogenous bioscavenger enzymes frommice, the median lethal dose for several OP nerve agents in Es1 KO mice was significantly lower than that in wild type control mice. Physiological and behavioral characterizations of these mice have been conducted in an effort to determine their suitability as a small animal model for OP nerve agentintoxication.Removal of serum carboxylesterasere presents one piece of a gestalt in vivo model not only for predicting human OP nerve agent susceptibility but also for testing medical countermeasures for that intoxication. One of the primary molecular targets of many current and novel OP countermeasures isacetylcholinesterase (AChE), the enzyme which hydrolyzes the neurotransmitter acetylcholineto terminate nerve signaling. When the active site of this enzyme is covalently inhibited by OP nerve agents, the resulting cholinergic crisis can quickly result in death. Several countermeasures have been developed which act directly upon inhibited AChE to remove the OP nerve agent and restore enzyme activity. While AChE is found in all species, significant biochemical differences resulting from species‐specific amino acid variations have been observed in a variety of in vitro experiments. Efforts to quantify these differences using in vitro models have been undertaken in this study.Support or Funding Information*This research was supported in part by an appointment to the Postgraduate Research Participation Program at the U.S. Army Medical Research Institute of Chemical Defense administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the U.S. Department of Energy and the U. S. Army Medical Research and Materiel Command. The experimental protocol was approved by the Animal Care and Use Committee at the United States Army Medical Research Institute of Chemical Defense and all procedures were conducted in accordance with the principles stated in the Guide for the Care and Use of Laboratory Animals (National Research Council, 2011), and the Animal Welfare Act of 1966 (P.L. 89‐544), as amended. This research was supported by the Defense Threat Reduction Agency – Joint Science and Technology Office, Medical S&T Division. The views expressed in this abstract are those of the author(s) and do not reflect the official policy of the Department of Army, Department of Defense, or the U.S. Government.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.