Long-term in vivo expression of a broad and potent entry inhibitor could circumvent the need for a conventional vaccine for HIV-1. Adeno-associated virus (AAV) vectors can stably express HIV-1 broadly neutralizing antibodies (bNAbs)1,2. However even the best bNAbs neutralize 10–50% of HIV-1 isolates inefficiently (IC80 > 5 μg/ml), suggesting that high concentrations of these antibodies would be necessary to achieve general protection3–6. Here we show that eCD4-Ig, a fusion of CD4-Ig with a small CCR5-mimetic sulfopeptide, binds avidly and cooperatively to the HIV-1 envelope glycoprotein (Env) and is more potent than the best bNAbs (geometric mean IC50 < 0.05 μg/ml). Because eCD4-Ig binds only conserved regions of Env, it is also much broader than any bNAb. For example, eCD4-Ig efficiently neutralized 100% of a diverse panel of neutralization-resistant HIV-1, HIV-2, and SIV isolates, including a comprehensive set of isolates resistant to the CD4-binding site bNAbs VRC01, NIH45-46, and 3BNC117. Rhesus macaques inoculated with an AAV vector stably expressed 17 to 77 μg/ml of fully functional rhesus eCD4-Ig for 40 weeks, and these macaques were protected from multiple infectious challenges with SHIV-AD8. Rhesus eCD4-Ig was also markedly less immunogenic than rhesus forms of four well characterized bNAbs. Our data suggest that AAV-delivered eCD4-Ig can function like an effective HIV-1 vaccine.

Highlights d Microglia engulf and eliminate synapses in the visual thalamus of MS patients d MS-relevant animal models show synapse engulfment and loss occur early in disease d Complement C3, but not C1q, localizes to synapses in demyelinating disease d AAV-Crry inhibits C3 and microglia-mediated synapse loss and preserves function

The cJun NH2-terminal kinase (JNK) stress signaling pathway is implicated in the metabolic response to the consumption of a high fat diet, including the development of obesity and insulin resistance. These metabolic adaptations involve altered liver function. Here we demonstrate that hepatic JNK potently represses the nuclear hormone receptor peroxisome proliferator-activated receptor α (PPARα). JNK therefore causes decreased expression of PPARα target genes that increase fatty acid oxidation / ketogenesis and promote the development of insulin resistance. We show that the PPARα target gene fibroblast growth factor 21 (Fgf21) plays a key role in this response because disruption of the hepatic PPARα - FGF21 hormone axis suppresses the metabolic effects of JNK-deficiency. This analysis identifies the hepatokine FGF21 as a critical mediator of JNK signaling in the liver.

Increased anxiety is a predominant withdrawal symptom in abstinent smokers, yet the neuroanatomical and molecular bases underlying it are unclear. Here, we show that withdrawal-induced anxiety increases activity of neurons in the interpeduncular intermediate (IPI), a subregion of the interpeduncular nucleus (IPN). IPI activation during nicotine withdrawal was mediated by increased corticotropin releasing factor (CRF) receptor-1 expression and signaling, which modulated glutamatergic input from the medial habenula (MHb). Pharmacological blockade of IPN CRF1 receptors or optogenetic silencing of MHb input reduced IPI activation and alleviated withdrawal-induced anxiety; whereas IPN CRF infusion in mice increased anxiety. We identified a meso-interpeduncular circuit, consisting of ventral tegmental area (VTA) dopaminergic neurons projecting to the IPN, as a potential source of CRF. Knock-down of CRF synthesis in the VTA prevented IPI activation and anxiety during nicotine withdrawal. These data indicate that increased CRF receptor signaling within a VTA-IPN-MHb circuit triggers anxiety during nicotine withdrawal.

Rhesus and cynomolgus macaques were challenged with 107 PFU of a clinical isolate of the severe acute respiratory syndrome (SARS) coronavirus. Some of the animals developed a mild self-limited respiratory infection very different from that observed in humans with SARS. The macaque model as it currently exists will have limited utility in the study of SARS and the evaluation of therapies.A novel coronavirus was recently identified as the causative agent of severe acute respiratory syndrome (SARS) (3, 7). One factor that led to this conclusion was the demonstration by Fouchier et al. and Kuiken et al. that exposure of the SARS coronavirus (SARS CoV) to a related host (i.e., cynomolgus macaques) led to the development of a comparable disease (1, 4). In their studies, SARS CoV administered intratracheally to effect delivery to the lower respiratory tract and conjunctiva of cynomolgus macaques resulted in respiratory and constitutional symptoms, shedding of virus, and pulmonary pathology. The availability of an authentic animal model for SARS is essential for a greater understanding of pathogenesis as well as the development and evaluation of effective vaccines and pharmacologic therapies.Our studies included a 12-to 14-day life phase of evaluation with sequential clinical and virologic analyses, followed by a thorough evaluation of tissues harvested at necropsy. This differed from the previous studies, which emphasized short-term necropsies (i.e., 4 to 6 days) and end points of pathology. Individually housed macaques were acclimated in a biosafety level 3 facility prior to initiation of the experiment.Cynomolgus macaques (Philippine origin and captive bred, 1.5 to 2.0 kg) and rhesus macaques (Indian origin and captive bred, 2.9 to 4.9 kg) were administered 10 7 PFU of Tor2 SARS CoV by direct instillation into the trachea via an endotracheal tube or intravenous infusion via a saphenous vein. These experiments were approved by the Animal Care and Use Committees of the University of Pennsylvania and the Southern Research Institute. All in vivo and in vitro experiments with SARS CoV were performed under biosafety level 3 with investigators using respirators. The challenge stock was 1 passage removed from the passage 2 stock provided to us from Heinz Feldmann (Winnipeg, Canada). An aliquot of the passage stock was converted to DNA with reverse transcriptase (RT), and open reading frames spanning N, M, E, and S were PCR amplified and subcloned, and one clone from each open reading frame was sequenced (both strands to 99.9% confidence level). The nucleotide sequences of the cloned open reading frames were 99.9% identical to the published Tor2 sequence (5). The abundance of SARS CoV RNA was quantitated in tissues by TaqMan PCR using oligonucleotides or probe to N. To minimize cross-contamination between samples during the necropsy, disposable tools were used for tissue and sample processing. In the TaqMan assay, cloned SARS CoV gene fragments were used to establish standard curves. Samples not treated with RT were incorpora...