Fusion of Lysostaphin to an Albumin Binding Domain Prolongs Its Half-Life and Bactericidal Activity in the Systemic Circulation

Antibacterial lysins are promising proteins that are active against both antibiotic-susceptible and antibiotic-resistant bacterial strains. However, a major limitation of antibacterial lysins is their fast elimination from systemic circulation. PEGylation increases the plasma half-life of lysins but renders them inactive. Here we report the construction of a fusion protein of lysostaphin, a potent anti-staphylococcal lysin, and an albumin-binding domain from streptococcal protein G. The resulting fusion protein was less active than the parent enzyme lysostaphin, but it still retained significant antibacterial activity even when bound to serum albumin. The terminal half-life of the fusion protein in rats was five-fold greater than that of lysostaphin (7.4 vs. 1.5 h), and the area under the curve increased more than 115 times. Most importantly, this increase in systemic circulation time compensated for the decrease in activity. The plasma from rats that received an injection of the fusion protein retained bactericidal activity for up to 7 h, while plasma from rats that received plain lysostaphin lacked any detectable activity after 4 h. To the best of our knowledge, this is the first report of an antibacterial lysin with both improved pharmacokinetic parameters and prolonged bactericidal activity in the systemic circulation

The Influence of Dimerization on the Pharmacokinetics and Activity of an Antibacterial Enzyme Lysostaphin

The increasing prevalence of antibiotic-resistant strains of pathogenic bacteria is a major healthcare problem. Antibacterial lysins are enzymes that cleave the peptidoglycan of the bacterial cell wall. These proteins hold potential as a supplement or an alternative to traditional antibiotics since they are active against antibiotic resistant strains. However, antibacterial lysins are rapidly eliminated from the systemic circulation, which limits their application. Dimerization of an anti-pneumococcal lysin Cpl-1 has been demonstrated to decrease the clearance rate of this protein in mice. In the present work, we constructed a dimer of an anti-staphylococcal lysin lysostaphin by fusing it with an anti-parallel α-helical dimerization domain. Lysostaphin dimer had a more favorable pharmacokinetic profile with increased terminal half-life and area under the curve (AUC) values compared to monomeric lysostaphin. However, the staphylolytic activity of dimerized lysostaphin was decreased. This decrease in activity was likely caused by the dimerization; since the catalytic efficacy of lysostaphin dimer towards pentaglycine peptide was unaltered. Our results demonstrate that, although dimerization is indeed beneficial for the pharmacokinetics of antibacterial lysins, this approach might not be suitable for all lysins, as it can negatively affect the lysin activity

Stimulation of Dectin-1 and Dectin-2 during Parenteral Immunization, but Not Mincle, Induces Secretory IgA in Intestinal Mucosa

Induction of a robust and long-lived mucosal immune response during vaccination is critical to achieve protection against numerous pathogens. However, traditional injected vaccines are generally poor inducers of mucosal immunity. One of the effective strategies to improve vaccine efficacy is incorporation of adjuvant molecules that enhance and polarize adaptive immune reactions. Effects of Syk-coupled lectin receptor agonists as adjuvants to induce mucosal immune reactions during parenteral immunization are not fully studied. We now report that the agonists trehalose-6,6-dibehenate (TDB), curdlan, and furfurman, which stimulate Dectin-1, Dectin-2, and Mincle, respectively, activate transcription factors (NF-κB, NFAT, and AP-1) to various extents in murine RAW 264.7 macrophages, even though similar pathways are activated. The agonists also elicit differential expression of maturation markers in bone marrow-derived dendritic cells, as well as differential cytokine secretion from these cells and from splenic mononuclear cells. In vivo assays also show that agonists of Dectin-1 and Dectin-2, but not Mincle, induce heavy IgA secretion in intestinal mucosa even when delivered parenterally. Strikingly, this effect appears to be formulation-independent. Collectively, the data suggest that adjuvants based on Dectin-1 and Dectin-2 agonists may significantly improve the efficacy of parenteral vaccines by inducing robust local immune reactions in intestinal mucosa

Model-Independent Observation of Exotic Contributions to B0→J/ψK+π−B^0\to J/\psi K^+\pi^- Decays

International audienceAn angular analysis of B0→J/ψK+π- decays is performed, using proton-proton collision data corresponding to an integrated luminosity of 3  fb-1 collected with the LHCb detector. The m(K+π-) spectrum is divided into fine bins. In each m(K+π-) bin, the hypothesis that the three-dimensional angular distribution can be described by structures induced only by K* resonances is examined, making minimal assumptions about the K+π- system. The data reject the K*-only hypothesis with a large significance, implying the observation of exotic contributions in a model-independent fashion. Inspection of the m(J/ψπ-) vs m(K+π-) plane suggests structures near m(J/ψπ-)=4200 and 4600 MeV

Study of Υ\Upsilon production in ppPb collisions at sNN=8.16\sqrt{s_{NN}}=8.16 TeV

International audienceThe production of ϒ(nS) mesons (n = 1, 2, 3) in pPb and Pbp collisions at a centre-of-mass energy per nucleon pair $\sqrt{s_{\mathrm{NN}}}=8.16$ TeV is measured by the LHCb experiment, using a data sample corresponding to an integrated luminosity of 31.8 nb$^{−1}$. The ϒ(nS) mesons are reconstructed through their decays into two opposite-sign muons. The measurements comprise the differential production cross-sections of the ϒ(1S) and ϒ(2S) states, their forward-to-backward ratios and nuclear modification factors. The measurements are performed as a function of the transverse momentum p$_{T}$ and rapidity in the nucleon-nucleon centre-of-mass frame y$^{*}$ of the ϒ(nS) states, in the kinematic range p$_{T}$ < 25 GeV/c and 1.5 < y$^{*}$ < 4.0 (−5.0 < y$^{*}$ < −2.5) for pPb (Pbp) collisions. In addition, production cross-sections for ϒ(3S) are measured integrated over phase space and the production ratios between all three ϒ(nS) states are determined. Suppression for bottomonium in proton-lead collisions is observed, which is particularly visible in the ratios. The results are compared to theoretical models

Measurement of the branching fraction and CPCP asymmetry in B+→J/ψρ+B^{+}\rightarrow J/\psi \rho^{+} decays

International audienceThe branching fraction and direct $C\!P$ asymmetry of the decay ${{{B} ^+}} \!\rightarrow {{J /\psi }} {{\rho } ^+}$ are measured using proton-proton collision data collected with the LHCb detector at centre-of-mass energies of 7 and 8 TeV, corresponding to a total integrated luminosity of 3 $\,\text{ fb }^{-1}$ . The following results are obtained: $\begin{aligned} \mathcal {B}({{B} ^+} \!\rightarrow {{J /\psi }} {{\rho } ^+} )&= (3.81^{+0.25-0.24} \pm 0.35) \times 10^{-5},\\ \mathcal {A}^{{C\!P}} ({{B} ^+} \!\rightarrow {{J /\psi }} {{\rho } ^+} )&= -0.045^{+0.056-0.057} \pm 0.008, \end{aligned}$ where the first uncertainties are statistical and the second systematic. Both measurements are the most precise to date

Amplitude analysis of the B(s)0→K∗0K‾∗0B^0_{(s)} \to K^{*0} \overline{K}^{*0} decays and measurement of the branching fraction of the B0→K∗0K‾∗0B^0 \to K^{*0} \overline{K}^{*0} decay

International audienceThe ${B}^0\to {K}^{\ast 0}{\overline{K}}^{\ast 0}$ and ${B}_s^0\to {K}^{\ast 0}{\overline{K}}^{\ast 0}$ decays are studied using proton-proton collision data corresponding to an integrated luminosity of 3 fb$^{−1}$. An untagged and timeintegrated amplitude analysis of B_{( s}_{)}^{0}  → (K$^{+}$π$^{−}$)(K$^{−}$π$^{+}$) decays in two-body invariant mass regions of 150 MeV/c$^{2}$ around the K$^{∗0}$ mass is performed. A stronger longitudinal polarisation fraction in the ${B}^0\to {K}^{\ast 0}{\overline{K}}^{\ast 0}$ decay, f$_{L}$ = 0.724 ± 0.051 (stat) ± 0.016 (syst), is observed as compared to f$_{L}$ = 0.240 ± 0.031 (stat) ± 0.025 (syst) in the ${B}_s^0\to {K}^{\ast 0}{\overline{K}}^{\ast 0}$ decay. The ratio of branching fractions of the two decays is measured and used to determine $\mathrm{\mathcal{B}}\left({B}^0\to {K}^{\ast 0}{\overline{K}}^{\ast 0}\right)=\left(8.0\pm 0.9\left(\mathrm{stat}\right)\pm 0.4\left(\mathrm{syst}\right)\right)\times {10}^{-7}$

Measurement of CPCP-violating and mixing-induced observables in Bs0→ϕγB_s^0 \to \phi\gamma decays

International audienceA time-dependent analysis of the Bs0→ϕγ decay rate is performed to determine the CP -violating observables Sϕγ and Cϕγ and the mixing-induced observable AϕγΔ. The measurement is based on a sample of pp collision data recorded with the LHCb detector, corresponding to an integrated luminosity of 3  fb-1 at center-of-mass energies of 7 and 8 TeV. The measured values are Sϕγ=0.43±0.30±0.11, Cϕγ=0.11±0.29±0.11, and AϕγΔ=-0.67-0.41+0.37±0.17, where the first uncertainty is statistical and the second systematic. This is the first measurement of the observables S and C in radiative Bs0 decays. The results are consistent with the standard model predictions