The rockefeller university

Broadly neutralizing antibodies (bNAbs) against HIV-1 provide both effective pre-exposure prophylaxis and treatment of HIV-1 infection in murine and nonhuman primate models, suggesting their potential use in humans. Although much is known about the role of variable domains in the neutralization breadth and potency of these bNAbs, the contribution of Fc domains to their activities is, by contrast, poorly characterized. Assessment of the in vivo activity of several bNAbs revealed that Fc gamma R-mediated effector function contributes substantially to their capacity to block viral entry, suppress viremia, and confer therapeutic activity. Enhanced in vivo potency of anti-HIV-1 bNAbs was associated with preferential engagement of activating, but not inhibitory Fc gamma Rs, and Fc domain-engineered bNAb variants with selective binding capacity for activating Fc gamma Rs displayed augmented protective activity. These findings reveal key roles for Fc effector function in the in vivo activity of anti-HIV-1 bNAbs and provide strategies for generating bNAbs with improved efficacy.

It is widely appreciated that effective human vaccines directed against viral pathogens elicit neutralizing antibodies (NAbs). The passive transfer of anti-HIV-1 NAbs conferring sterilizing immunity to macaques has been used to determine the plasma neutralization titers, which must be present at the time of exposure, to prevent acquisition of SIV/HIV chimeric virus (SHIV) infections. We administered five recently isolated potent and broadly acting anti-HIV neutralizing monoclonal antibodies (mAbs) to rhesus macaques and challenged them intrarectally 24 h later with either of two different R5-tropic SHIVs. By combining the results obtained from 60 challenged animals, we determined that the protective neutralization titer in plasma preventing virus infection in 50% of the exposed monkeys was relatively modest (similar to 1:100) and potentially achievable by vaccination.

Mycosis fungoides, the most common type of cutaneous T-cell lymphoma (CTCL), is characterized by a helper T-cell 2 (Th2) skewing with a mature CD4(+) memory T-cell phenotype. Using skin samples from patients with mycosis fungoides (n = 21), healthy volunteers (n = 17), and individuals with atopic dermatitis (n = 17) and psoriasis (n = 9), we found IL32 mRNA expression significantly higher in mycosis fungoides samples than in samples from benign inflammatory skin diseases, and its expression increases with disease progression. By IHC and immunofluorescence, we confirmed IL32 protein expression in many CD3(+)CD4(+)T cells and some epidermotropic T cells in mycosis fungoides lesions. MyLa cells (a mycosis fungoides cell line) express IL32, which, in turn, could promote cellular proliferation and viability in a dose-dependent fashion. IL32-treated MyLa and CTCL HH cells upregulated cell proliferation and survival genes. Of the major "polarizing" T-cell cytokines, only IFN gamma mRNA increases with mycosis fungoides progression and positively correlates with IL32 mRNA expression. Th2 cytokines do not positively correlate with IL32 mRNA expression or mycosis fungoides progression. Furthermore, by flow cytometry, IL32 production by circulating activated T cells in healthy individuals was found in both IFN gamma(+) and IFN gamma(-) cells but not in IL4(+)or IL13(+) cells. In conclusion, we have identified IL32(+) cells as the likely tumor cells in mycosis fungoides, and demonstrated that IL32 mRNA expression increases with mycosis fungoides progression and is significantly higher than mRNA expression in other skin diseases, and that some IL32(+) T cells are independent from the defined Th subsets. Thus, IL32 may play a unique role in mycosis fungoides progression as an autocrine cytokine. (C)2014 AACR.

Searches for the direct electroweak production of supersymmetric charginos, neutralinos, and sleptons in a variety of signatures with leptons and W, Z, and Higgs bosons are presented. Results are based on a sample of proton-proton collision data collected at center-of-mass energy root s = 8 TeV with the CMS detector in 2012, corresponding to an integrated luminosity of 19.5 fb(-1). The observed event rates are in agreement with expectations from the standard model. These results probe charginos and neutralinos with masses up to 720 GeV, and sleptons up to 260 GeV, depending on the model details.

A search for supersymmetry through the direct pair production of top squarks, with Higgs (H) or Z bosons in the decay chain, is performed using a data sample of proton-proton collisions at root s = 8 TeVcollected in 2012 with the CMS detector at the LHC. The sample corresponds to an integrated luminosity of 19.5 fb(-1). The search is performed using a selection of events containing leptons and bottom-quark jets. No evidence for a significant excess of events over the standard model background prediction is observed. The results are interpreted in the context of simplified supersymmetric models with pair production of a heavier top-squark mass eigenstate (t) over tilde (2) decaying to a lighter top-squark mass eigenstate (t) over tilde (1) via either (t) over tilde (2) -> H (t) over tilde (1) or (t) over tilde (2) -> Z (t) over tilde (1), followed in both cases by (t) over tilde (1) -> t (chi) over tilde (0)(1), where (chi) over tilde (0)(1) is an undetected, stable, lightest supersymmetric particle. The interpretation is performed in the region where the mass difference between the (t) over tilde (1) and (chi) over tilde (0)(1) states is approximately equal to the top-quark mass (m((t) over tilde )1) - m((chi) over tilde 10) similar or equal to m(t)) which is not probed by searches for direct (t) over tilde (1) squark pair production. The analysis excludes top squarks with masses m((t) over tilde2) < 575 GeVand m(<(t)over tilde>1) < 400 GeVat a 95% confidence level. (C) 2014 The Authors. Published by Elsevier B.V.

We have previously reported on a unique patient in whom homozygosity for a mutation at IRF8 (IRF8(K108E)) causes a severe immunodeficiency. Laboratory evaluation revealed a highly unusual myeloid compartment, remarkable for the complete absence of CD14(+) and CD16(+) monocytes, absence of CD11c(+) conventional dendritic cells (DCs) and CD11c(+)/CD123(+) plasmacytoid DCs, and striking granulocytic hyperplasia. The patient initially presented with severe disseminated mycobacterial and mucocutaneous fungal infections and was ultimately cured by cord blood transplant. Sequencing RNA from the IRF8(K108E) patient's primary blood cells prior to transplant shows not only depletion of IRF8-bound and IRF8-regulated transcriptional targets, in keeping with the distorted composition of the myeloid compartment, but also a paucity of transcripts associated with activated CD4(+) and CD8(+) T lymphocytes. This suggests that T cells reared in the absence of a functional antigen-presenting compartment in IRF8(K108E) are anergic. Biochemical characterization of the IRF8(K108E) mutant in vitro shows that loss of the positively charged side chain at K108 causes loss of nuclear localization and loss of transcriptional activity, which is concomitant with decreased protein stability, increased ubiquitination, increased small ubiquitin-like modification, and enhanced proteasomal degradation. These findings provide functional insight into the molecular basis of immunodeficiency associated with loss of IRF8.

Production of prompt J/psi meson pairs in proton-proton collisions at root s = 7 TeV is measured with the CMS experiment at the LHC in a data sample corresponding to an integrated luminosity of about 4.7 fb(-1). The two J/psi mesons are fully reconstructed via their decays into mu(+)mu(-) pairs. This observation provides for the first time access to the high-transverse-momentum region of J/psi pair production where model predictions are not yet established. The total and differential cross sections are measured in a phase space defined by the individual J/psi transverse momentum (p(T)(J/psi)) and rapidity (vertical bar y(J/psi)vertical bar): vertical bar y(J/psi)vertical bar < 1.2 for p(T)(J/psi) > 6.5 GeV/c; 1.2 < vertical bar y(J/psi)vertical bar < 1.43 for a pT threshold that scales linearly with vertical bar y(J/psi)vertical bar from 6.5 to 4.5 GeV/c; and 1.43 < vertical bar y(J/psi)vertical bar < 2.2 for p(T)(J/psi) > 4.5 GeV/c. The total cross section, assuming unpolarized prompt J/psi pair production is 1.49 +/- 0.07 (stat) +/- 0.13 (syst) nb. Different assumptions about the J/psi polarization imply modifications to the cross section ranging from -31% to +27%.

Objective-Aortic valve stenosis (AS) is characterized by fibrosis and calcification of valves leading to aortic valve narrowing, resulting in high wall shear stress across the valves. We previously demonstrated that high shear stress can activate platelet-derived transforming growth factor-beta 1 (TGF-beta 1), a cytokine that induces fibrosis and calcification. The aim of this study was to investigate the role of shear-induced platelet release of TGF-beta 1 and its activation in AS. Approach and Results-We studied hypercholesterolemic Ldlr(-/-)Apob(100/100)/Mttp(fl/fl)/Mx1Cre(+/+) (Reversa) mice that develop AS on Western diet and a surgical ascending aortic constriction mouse model that acutely simulates the hemodynamics of AS to study shear-induced platelet TGF-beta 1 release and activation. Reversa mice on Western diet for 6 months had thickening of the aortic valves, increased wall shear stress, and increased plasma TGF-beta 1 levels. There were weak and moderate correlations between wall shear stress and TGF-beta 1 levels in the progression and reversed Reversa groups and a stronger correlation in the ascending aortic constriction model in wild-type mice but not in mice with a targeted deletion of megakaryocyte and platelet TGF-beta 1 (Tgfb1f(lox)). Plasma total TGF-beta 1 levels correlated with collagen deposition in the stenotic valves in Reversa mice. Although active TGF-beta 1 levels were too low to be measured directly, we found (1) canonical TGF-beta 1 (phosphorylated small mothers against decapentaplegic 2/3) signaling in the leukocytes and canonical and noncanonical (phosphorylated extracellular signal-regulated kinases 1/2) TGF-beta 1 signaling in aortic valves of Reversa mice on a Western diet, and (2) TGF-beta 1 signaling of both pathways in the ascending aortic constriction stenotic area in wild-type but not Tgfb1f(lox) mice. Conclusions-Shear-induced, platelet-derived TGF-beta 1 activation may contribute to AS.

The transforming growth factor-beta (TGF-beta) pathway plays a fundamental role in development and disease. Despite its importance, the dynamics of signaling activity downstream of ligand stimulation have remained largely unexplored. The recent study by Vizan et al. demonstrates that loss of signaling-capable receptors from the cell surface leads to a refractory period during which cells are incapable of responding to additional signals. In this letter and in our previous work, we show that although receptor dynamics determine Smad2/3 activity, signaling activity at the level of transcription terminates far earlier in a receptor- and Smad2/3-independent manner. Thus, Smad2/3 activity does not reveal the dynamics of transcription, and downstream measures must be examined directly.

The circumsporozoite protein (CSP) of Plasmodium falciparum is a major surface protein, which forms a dense coat on the sporozoite's surface. Preclinical research on CSP and clinical evaluation of a CSP fragment-based RTS, S/AS01 vaccine have demonstrated a modest degree of protection against P. falciparum, mediated in part by humoral immunity and in part by cell-mediated immunity. Given the partial protective efficacy of the RTS, S/AS01 vaccine in a recent Phase 3 trial, further improvement of CSP-based vaccines is crucial. In this report, we describe the preclinical development of a full-length, recombinant CSP (rCSP)-based vaccine candidate against P. falciparum malaria suitable for current Good Manufacturing Practice (cGMP) production. Utilizing a novel high-throughput Pseudomonas fluorescens expression platform, we demonstrated greater efficacy of full-length rCSP as compared to N-terminally truncated versions, rapidly down-selected a promising lead vaccine candidate, and developed a high-yield purification process to express immunologically active, intact antigen for clinical trial material production. The rCSP, when formulated with various adjuvants, induced antigen-specific antibody responses as measured by enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assay (IFA), as well as CD4+ T-cell responses as determined by ELISpot. The adjuvanted rCSP vaccine conferred protection in mice when challenged with transgenic P. berghei sporozoites containing the P. falciparum repeat region of CSP. Furthermore, heterologous prime/boost regimens with adjuvanted rCSP and an adenovirus type 35-vectored CSP (Ad35CS) showed modest improvements in eliciting CSP-specific T-cell responses and anti-malarial protection, depending on the order of vaccine delivery. Collectively, these data support the importance of further clinical development of adjuvanted rCSP, either as a stand-alone product or as one of the components in a heterologous prime/boost strategy, ultimately acting as an effective vaccine candidate for the mitigation of P. falciparum-induced malaria.