Enforced Expression of the Transcriptional Coactivator OBF1 Impairs B Cell Differentiation at the Earliest Stage of Development

OBF1, also known as Bob.1 or OCA-B, is a B lymphocyte-specific transcription factor which coactivates Oct1 and Oct2 on B cell specific promoters. So far, the function of OBF1 has been mainly identified in late stage B cell populations. The central defect of OBF1 deficient mice is a severely reduced immune response to T cell-dependent antigens and a lack of germinal center formation in the spleen. Relatively little is known about a potential function of OBF1 in developing B cells. Here we have generated transgenic mice overexpressing OBF1 in B cells under the control of the immunoglobulin heavy chain promoter and enhancer. Surprisingly, these mice have greatly reduced numbers of follicular B cells in the periphery and have a compromised immune response. Furthermore, B cell differentiation is impaired at an early stage in the bone marrow: a first block is observed during B cell commitment and a second differentiation block is seen at the large preB2 cell stage. The cells that succeed to escape the block and to differentiate into mature B cells have post-translationally downregulated the expression of transgene, indicating that expression of OBF1 beyond the normal level early in B cell development is deleterious. Transcriptome analysis identified genes deregulated in these mice and Id2 and Id3, two known negative regulators of B cell differentiation, were found to be upregulated in the EPLM and preB cells of the transgenic mice. Furthermore, the Id2 and Id3 promoters contain octamer-like sites, to which OBF1 can bind. These results provide evidence that tight regulation of OBF1 expression in early B cells is essential to allow efficient B lymphocyte differentiation

The regulated long-term delivery of therapeutic proteins using antigen-specific B lymphocytes

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Early embryonic expression patterns of the mouse Flamingo and Prickle orthologues

The Drosophila melanogaster proteins Flamingo and Prickle act in the planar cell polarity (PCP) pathway, which is required for acquisition of epithelial polarity in the wing, eye, and epidermis. In mammals, PCP signaling has been shown to regulate cell movements and polarity in a variety of tissues. Here, we show that the murine Flamingo orthologues Celsr1-3 and the Prickle orthologues Prickle1, Prickle2, and Testin have dynamic patterns of expression during pregastrulation and gastrulation stages. Celsr1 is expressed in the anterior visceral endoderm and nascent mesoderm, Celsr2 and Celsr3 mark the prospective neuroectoderm, Prickle1 is expressed in the primitive streak and mesoderm, Prickle2 in the node, and Testin in the anterior visceral endoderm, the extraembryonic ectoderm, primitive streak, and mesoderm. Analysis of a gene-trap mutation in Testin indicates that this gene is not required for embryogenesis; therefore, other Prickle homologues may compensate for its function during development

Fiber Buckling in Confined Viscous Flows: An Absolute Instability Described by the Linear Ginzburg-Landau Equation

International audienceWe explore the dynamics of a flexible fiber transported by a viscous flow in a Hele-Shaw cell of height comparable to the fiber height. We show that long fibers aligned with the flow experience a buckling instability. Competition between viscous and elastic forces leads to the deformation of the fiber into a wavy shape convolved by a Bell-shaped envelope. We characterize the wavelength and phase velocity of the deformation as well as the growth and spreading of the envelope. Our study of the spatiotemporal evolution of the deformation reveals a linear and absolute instability arising from a local mechanism well described by the Ginzburg-Landau equation

Microfluidic in situ mechanical testing of photopolymerized gels

International audienceGels are a functional template for micro-particle fabrication and microbiology experiments. The control and knowledge of their mechanical properties is critical in a number of applications, but no simple in situ method exists to determine these properties. We propose a novel microfluidic based method that directly measures the mechanical properties of the gel upon its fabrication. We measure the deformation of a gel beam under a controlled flow forcing, which gives us a direct access to the Young's modulus of the material itself. We then use this method to determine the mechanical properties of poly(ethylene glycol) diacrylate (PEGDA) under various experimental conditions. The mechanical properties of the gel can be highly tuned, yielding two order of magnitude in the Young's modulus. The method can be easily implemented to allow for an in situ direct measurement and control of Young's moduli under various experimental conditions

Fiber Buckling in Confined Viscous Flows: An Absolute Instability Described by the Linear Ginzburg-Landau Equation

International audienceWe explore the dynamics of a flexible fiber transported by a viscous flow in a Hele-Shaw cell of height comparable to the fiber height. We show that long fibers aligned with the flow experience a buckling instability. Competition between viscous and elastic forces leads to the deformation of the fiber into a wavy shape convolved by a Bell-shaped envelope. We characterize the wavelength and phase velocity of the deformation as well as the growth and spreading of the envelope. Our study of the spatiotemporal evolution of the deformation reveals a linear and absolute instability arising from a local mechanism well described by the Ginzburg-Landau equation

Transport of flexible fibers in confined microchannels

When transported in confined geometries rigid fibers show interesting transport dynamics induced by friction with the top and bottom walls. Fiber flexibility causes an additional coupling between fiber deformation and transport and is expected to lead to more complex dynamics. A first crucial step for their understanding is the characterization of the deformed fiber shape. Here we characterize this shape for a fiber transported in a confined plug flow perpendicular to the flow direction using a combination of microfluidic experiments and numerical simulations. In the experiments, size, initial orientation, and mechanical properties of the fibers are controlled using microfabrication techniques and in situ characterization methods. The numerical simulations use modified Brinkman equations as well as full three-dimensional simulations. We show that the bending of a perpendicular fiber results from the force distribution acting on the elongated object and is proportional to the elasto-viscous number, which compares viscous to elastic forces. We quantitatively characterize the influence of the confinement on the fiber deformation. The precise understanding of the deformation of a flexible fiber in a confined geometry can also be used in future to understand the deformation and transport of more complex deformable particles in confined flows, such as vesicles or red blood cells

Deformation of a flexible fiber settling in a quiescent viscous fluid

International audienceThe equilibrium state of a flexible fiber settling in a viscous fluid is examined using a combination of macroscopic experiments, numerical simulations, and scaling arguments. We identify three regimes having different signatures on this equilibrium configuration of the elastic filament: weak and large deformation regimes wherein the drag is proportional to the settling velocity as expected in Stokes flow and an intermediate elastic reconfiguration regime where the filament deforms to adopt a shape with a smaller drag which is no longer linearly proportional to the velocity