Mouse model for acute Epstein-Barr virus infection

Epstein-Barr Virus (EBV) infects human B cells and drives them into continuous proliferation. Two key viral factors in this process are the latent membrane proteins LMP1 and LMP2A, which mimic constitutively activated CD40 receptor and B-cell receptor signaling, respectively. EBV-infected B cells elicit a powerful T-cell response that clears the infected B cells and leads to life-long immunity. Insufficient immune surveillance of EBV-infected B cells causes life-threatening lymphoproliferative disorders, including mostly germinal center (GC)-derived B-cell lymphomas. We have modeled acute EBV infection of naive and GC B cells in mice through timed expression of LMP1 and LMP2A. Although lethal when induced in all B cells, induction of LMP1 and LMP2A in just a small fraction of naive B cells initiated a phase of rapid B-cell expansion followed by a proliferative T-cell response, clearing the LMP-expressing B cells. Interfering with T-cell activity prevented clearance of LMP-expressing B cells. This was also true for perforin deficiency, which in the human causes a life-threatening EBV-related immunoproliferative syndrome. LMP expression in GC B cells impeded the GC reaction but, upon loss of T-cell surveillance, led to fatal B-cell expansion. Thus, timed expression of LMP1 together with LMP2A in subsets of mouse B cells allows one to study major clinically relevant features of human EBV infection in vivo, opening the way to new therapeutic approaches

Level density of a Fermi gas: average growth and fluctuations

We compute the level density of a two--component Fermi gas as a function of the number of particles, angular momentum and excitation energy. The result includes smooth low--energy corrections to the leading Bethe term (connected to a generalization of the partition problem and Hardy--Ramanujan formula) plus oscillatory corrections that describe shell effects. When applied to nuclear level densities, the theory provides a unified formulation valid from low--lying states up to levels entering the continuum. The comparison with experimental data from neutron resonances gives excellent results.Comment: 4 pages, 1 figur

Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes

BACKGROUND: The CRISPR/Cas9 system is increasingly used for gene inactivation in mouse zygotes, but homology-directed mutagenesis and use of inbred embryos are less established. In particular, Rosa26 knock-in alleles for the insertion of transgenes in a genomic 'safe harbor' site, have not been produced. Here we applied CRISPR/Cas9 for the knock-in of 8-11 kb inserts into Rosa26 of C57BL/6 zygotes. RESULTS: We found that 10-20 % of live pups derived from microinjected zygotes were founder mutants, without apparent off-target effects, and up to 50 % knock-in embryos were recovered upon coinjection of Cas9 mRNA and protein. Using this approach, we established a new mouse line for the Cre/loxP-dependent expression of Cas9. CONCLUSIONS: Altogether, our protocols and resources support the fast and direct generation of new Rosa26 knock-in alleles and of Cas9-mediated in vivo gene editing in the widely used C57BL/6 inbred strain

Efficient CRISPR/Cas9-mediated gene knockin in mouse hematopoietic stem and progenitor cells

Mutations accumulating in hematopoietic stem and progenitor cells (HSPCs) during development can cause severe hematological disorders. Modeling these mutations in mice is essential for understanding their functional consequences. Here, we describe an efficient CRISPR/Cas9-based system to knock in and repair genes in mouse HSPCs. CRISPR/Cas9 ribonucleoproteins, in combination with recombinant adeno-associated virus (rAAV)-DJ donor templates, led to gene knockin efficiencies of up to 30% in the Lmnb1 and Actb loci of mouse HSPCs in vitro. The targeted HSPCs engraft and reconstitute all immune cell lineages in the recipient mice. Using this approach, we corrected a neomycin-disrupted Rag2 gene. The Rag2-corrected HSPCs restore B and T cell development in vivo, confirming the functionality of the approach. Our method provides an efficient strategy to study gene function in the hematopoietic system and model hematological disorders in vivo, without the need for germline mutagenesis

Two Skyrmion Dynamics with Omega Mesons

We present our first results of numerical simulations of two skyrmion dynamics using an $\omega$-meson stabilized effective Lagrangian. We consider skyrmion-skyrmion scattering with a fixed initial velocity of $\beta=0.5$, for various impact parameters and groomings. The physical picture that emerges is surprisingly rich, while consistent with previous results and general conservation laws. We find meson radiation, skyrmion scattering out of the scattering plane, orbiting and capture to bound states.Comment: 19 pages, 22 figure

Symmetry Decomposition of Potentials with Channels

We discuss the symmetry decomposition of the average density of states for the two dimensional potential $V=x^2y^2$ and its three dimensional generalisation $V=x^2y^2+y^2z^2+z^2x^2$. In both problems, the energetically accessible phase space is non-compact due to the existence of infinite channels along the axes. It is known that in two dimensions the phase space volume is infinite in these channels thus yielding non-standard forms for the average density of states. Here we show that the channels also result in the symmetry decomposition having a much stronger effect than in potentials without channels, leading to terms which are essentially leading order. We verify these results numerically and also observe a peculiar numerical effect which we associate with the channels. In three dimensions, the volume of phase space is finite and the symmetry decomposition follows more closely that for generic potentials --- however there are still non-generic effects related to some of the group elements

B-cell-specific Myd88 L252P expression causes a premalignant gammopathy resembling IgM MGUS

A highly recurrent somatic L265P mutation in the TIR domain of the signaling adapter MYD88 constitutively activates NF-κB. It occurs in nearly all human patients with Waldenström's macroglobulinemia (WM), a B cell malignancy caused by IgM-expressing cells. Here, we introduced an inducible leucine to proline point mutation into the mouse Myd88 locus, at the orthologous position L252P. When the mutation was introduced early during B cell development, B cells developed normally. However, IgM-expressing plasma cells accumulated with age in spleen and bone, leading to more than 20-fold elevated serum IgM titers. When introduced into germinal center B cells in the context of an immunization, the Myd88(L252P) mutation caused prolonged persistence of antigen-specific serum IgM and elevated numbers of antigen-specific IgM plasma cells. Myd88(L252P)-expressing B cells switched normally, but plasma cells expressing other immunoglobulin isotypes did not increase in numbers, implying that IgM expression may be required for the observed cellular expansion. In order to test whether the Myd88(L252P) mutation can cause clonal expansions, we introduced it into a small fraction of CD19-positive B cells. In this scenario, five out of five mice developed monoclonal IgM serum paraproteins accompanied by an expansion of clonally related plasma cells that expressed mostly hypermutated VDJ regions. Taken together, our data suggest that the Myd88(L252P) mutation is sufficient to promote aberrant survival and expansion of IgM-expressing plasma cells which in turn can cause IgM monoclonal gammopathy of undetermined significance (MGUS), the premalignant condition that precedes WM

Collision-Induced Decay of Metastable Baby Skyrmions

Many extensions of the standard model predict heavy metastable particles which may be modeled as solitons (skyrmions of the Higgs field), relating their particle number to a winding number. Previous work has shown that the electroweak interactions admit processes in which these solitons decay, violating standard model baryon number. We motivate the hypothesis that baryon-number-violating decay is a generic outcome of collisions between these heavy particles. We do so by exploring a 2+1 dimensional theory which also possesses metastable skyrmions. We use relaxation techniques to determine the size, shape and energy of static solitons in their ground state. These solitons could decay by quantum mechanical tunneling. Classically, they are metastable: only a finite excitation energy is required to induce their decay. We attempt to induce soliton decay in a classical simulation by colliding pairs of solitons. We analyze the collision of solitons with varying inherent stabilities and varying incident velocities and orientations. Our results suggest that winding-number violating decay is a generic outcome of collisions. All that is required is sufficient (not necessarily very large) incident velocity; no fine-tuning of initial conditions is required.Comment: 24 pages, 7 figures, latex. Very small changes onl

Hartree Fock Calculations in the Density Matrix Expansion Approach

The density matrix expansion is used to derive a local energy density functional for finite range interactions with a realistic meson exchange structure. Exchange contributions are treated in a local momentum approximation. A generalized Slater approximation is used for the density matrix where an effective local Fermi momentum is chosen such that the next to leading order off-diagonal term is canceled. Hartree-Fock equations are derived incorporating the momentum structure of the underlying finite range interaction. For applications a density dependent effective interaction is determined from a G-matrix which is renormalized such that the saturation properties of symmetric nuclear matter are reproduced. Intending applications to systems far off stability special attention is paid to the low density regime and asymmetric nuclear matter. Results are compared to predictions obtained from Skyrme interactions. The ground state properties of stable nuclei are well reproduced without further adjustments of parameters. The potential of the approach is further exemplified in calculations for A=100...140 tin isotopes. Rather extended neutron skins are found beyond 130Sn corresponding to solid layers of neutron matter surrounding a core of normal composition.Comment: Revtex, 29 pages including 14 eps figures, using epsfig.st

Semiclassical Trace Formulas for Noninteracting Identical Particles

We extend the Gutzwiller trace formula to systems of noninteracting identical particles. The standard relation for isolated orbits does not apply since the energy of each particle is separately conserved causing the periodic orbits to occur in continuous families. The identical nature of the particles also introduces discrete permutational symmetries. We exploit the formalism of Creagh and Littlejohn [Phys. Rev. A 44, 836 (1991)], who have studied semiclassical dynamics in the presence of continuous symmetries, to derive many-body trace formulas for the full and symmetry-reduced densities of states. Numerical studies of the three-particle cardioid billiard are used to explicitly illustrate and test the results of the theory.Comment: 29 pages, 11 figures, submitted to PR