Impaired precursor B cell differentiation in Bruton's tyrosine kinase-deficient mice

Bruton's tyrosine kinase (Btk) is a cytoplasmic signaling molecule that is crucial for precursor (pre-B) cell differentiation in humans. In this study, we show that during the transition of large cycling to small resting pre-B cells in the mouse, Btk-deficient cells failed to efficiently modulate the expression of CD43, surrogate L chain, CD2, and CD25. In an analysis of the kinetics of pre-B cell differentiation in vivo, Btk-deficient cells manifested a specific developmental delay within the small pre-B cell compartment of about 3 h, when compared with wild-type cells. Likewise, in in vitro bone marrow cultures, Btk-deficient large cycling pre-B cells showed increased IL-7 mediated expansion and reduced developmental progression into noncycling CD2(+)CD25(+) surrogate L chain-negative small pre-B cells and subsequently into Ig-positive B cells. Furthermore, the absence of Btk resulted in increased proliferative responses to IL-7 in recombination-activating gene-1-deficient pro-B cells. These findings identify a novel role for Btk in the regulation of the differentiation stage-specific modulation of IL-7 responsiveness in pro-B and pre-B cells. Moreover, our results show that Btk is critical for an efficient transit through the small pre-B cell compartment, thereby regulating cell surface phenotype changes during the developmental progression of cytoplasmic mu H chain expressing pre-B cells into immature IgM(+) B cells

Function of Bruton's tyrosine kinase during B cell development is partially independent of its catalytic activity

The Tec family member Bruton's tyrosine kinase (Btk) is a cytoplasmic protein tyrosine kinase that transduces signals from the pre-B and B cell receptor (BCR). Btk is involved in pre-B cell maturation by regulating IL-7 responsiveness, cell surface phenotype changes, and the activation of lambda L chain gene rearrangements. In mature B cells, Btk is essential for BCR-mediated proliferation and survival. Upon BCR stimulation, Btk is transphosphorylated at position Y551, which promotes its catalytic activity and subsequently results in autophosphorylation at position Y223 in the Src homology 3 domain. To address the significance of Y223 autophosphorylation and the requirement of enzymatic activity for Btk function in vivo, we generated transgenic mice that express the autophosphorylation site mutant Y223F and the kinase-inactive mutant K430R, respectively. We found that Y223 autophosphorylation was not required for the regulation of IL-7 responsiveness and cell surface phenotype changes in differentiating pre-B cells, or for peripheral B cell differentiation. However, expression of the Y223F-Btk transgene could not fully rescue the reduction of lambda L chain usage in Btk-deficient mice. In contrast, transgenic expression of kinase-inactive K430R-Btk completely reconstituted lambda usage in Btk-deficient mice, but the defective modulation of pre-B cell surface markers, peripheral B cell survival, and BCR-mediated NF-kappaB induction were partially corrected. From these findings, we conclude that: 1) autophosphorylation at position Y223 is not essential for Btk function in vivo, except for regulation of lambda L chain usage, and 2) during B cell development, Btk partially acts as an adapter molecule, independent of its catalytic activity

Recent progress on the manipulation of single atoms in optical tweezers for quantum computing

This paper summarizes our recent progress towards using single rubidium atoms trapped in an optical tweezer to encode quantum information. We demonstrate single qubit rotations on this system and measure the coherence of the qubit. We move the quantum bit over distances of tens of microns and show that the coherence is reserved. We also transfer a qubit atom between two tweezers and show no loss of coherence. Finally, we describe our progress towards conditional entanglement of two atoms by photon emission and two-photon interferences.Comment: Proceedings of the ICOLS07 conferenc

VAMP8-mediated NOX2 recruitment to endosomes is necessary for antigen release

Contains fulltext : 178043.pdf (publisher's version ) (Open Access)Cross-presentation of foreign antigen in major histocompatibility complex (MHC) class I by dendritic cells (DCs) requires activation of the NADPH-oxidase NOX2 complex. We recently showed that NOX2 is recruited to phagosomes by the SNARE protein VAMP8 where NOX2-produced reactive oxygen species (ROS) cause lipid oxidation and membrane disruption, promoting antigen translocation into the cytosol for cross-presentation. In this study, we extend these findings by showing that VAMP8 is also involved in NOX2 trafficking to endosomes. Moreover, we demonstrate in both human and mouse DCs that absence of VAMP8 leads to decreased ROS production, lipid peroxidation and antigen translocation, and that this impairs cross-presentation. In contrast, knockdown of VAMP8 did not affect recruitment of MHC class I and the transporter associated with antigen processing 1 (TAP1) to phagosomes, although surface levels of MHC class I were reduced. Thus, in addition to a secretory role, VAMP8-mediates trafficking of NOX2 to endosomes and phagosomes and this promotes induction of cytolytic T cell immune responses

Using animations to connect macroscopic, sub-microscopic, and symbolic representations of the world

PROBLEM Chemistry is considered a difficult subject mainly due to multiplicity of representations, i.e. the Johnstone’s triangle of macroscopic, sub-microscopic, and symbolic domains (Johnstone, 1982). When students learn chemistry, moving between these domains is a well-recognised challenge (Cardellini, 2012). PLAN Teaching resources must address all three domains. For example, sub-microscopic level can be represented by cartoons, macroscopic level by video recordings of laboratory experiments, and symbolic by chemical reactions and mathematical calculations. ACTION To address this challenge, we developed a series of animations to illustrate the chemical phenomena most difficult for students. They were accompanied with the audio and text narration designed to get students to address their assumptions and misconceptions and to assist hearing-impaired and non-English speakers. The animations were evaluated using engagement analytics, test results, and student/academic comments. REFLECTION To develop these animations, we combined evidence-based pedagogy and technology, to improve student learning and enrich student experience. Animations address shifting expectations of our students by providing them with learner-centred approaches to enhance learning engagement and impact. Teaching and learning with animations is flexible and self-paced

Tumor suppressor function of Bruton tyrosine kinase is independent of its catalytic activity

During B-cell development in the mouse, Bruton tyrosine kinase (Btk) and the adaptor protein SLP-65 (Src homology 2 [SH2] domain-containing leukocyte protein of 65 kDa) limit the expansion and promote the differentiation of pre-B cells. Btk is thought to mainly function by phosphorylating phospholipase Cgamma2, which is brought into close proximity of Btk by SLP-65. However, this model was recently challenged by the identification of a role for Btk as a tumor suppressor in the absence of SLP-65 and by the finding that Btk function is partially independent of its kinase activity. To investigate if enzymatic activity is critical for the tumor suppressor function of Btk, we crossed transgenic mice expressing the kinase-inactive K430R-Btk mutant onto a Btk/SLP-65 double-deficient background. We found that K430R-Btk expression rescued the severe developmental arrest at the pre-B-cell stage in Btk/SLP-65 double-deficient mice. Moreover, K430R-Btk co

Bruton's Tyrosine Kinase Cooperates with the B Cell Linker Protein SLP-65 as a Tumor Suppressor in Pre-B Cells

Expression of the pre-B cell receptor (pre-BCR) leads to activation of the adaptor molecule SLP-65 and the cytoplasmic kinase Btk. Mice deficient for one of these signaling proteins have an incomplete block in B cell development at the stage of large cycling pre-BCR+CD43+ pre-B cells. Our recent findings of defective SLP-65 expression in ∼50% of childhood pre-B acute lymphoblastic leukemias and spontaneous pre-B cell lymphoma development in SLP-65−/− mice demonstrate that SLP-65 acts as a tumor suppressor. To investigate cooperation between Btk and SLP-65, we characterized the pre-B cell compartment in single and double mutant mice, and found that the two proteins have a synergistic role in the developmental progression of large cycling into small resting pre-B cells. We show that Btk/SLP-65 double mutant mice have a dramatically increased pre-B cell tumor incidence (∼75% at 16 wk of age), as compared with SLP-65 single deficient mice (<10%). These findings demonstrate that Btk cooperates with SLP-65 as a tumor suppressor in pre-B cells. Furthermore, transgenic low-level expression of a constitutive active form of Btk, the E41K-Y223F mutant, prevented tumor formation in Btk/SLP-65 double mutant mice, indicating that constitutive active Btk can substitute for SLP-65 as a tumor suppressor

Ray chaos in optical cavities based upon standard laser mirrors

We present a composite optical cavity made of standard laser mirrors; the cavity consists of a suitable combination of stable and unstable cavities. In spite of its very open nature the composite cavity shows ray chaos, which may be either soft or hard, depending on the cavity configuration. This opens a new, convenient route for experimental studies of the quantum aspects of a chaotic wave field.Comment: 4 pages, 3 figures, 1 tabl

Generation of pulsed and continuous-wave squeezed light with 87^{87}Rb vapor

We present experimental studies on the generation of pulsed and continuous-wave squeezed vacuum via nonlinear rotation of the polarization ellipse in a $^{87}$Rb vapor. Squeezing is observed for a wide range of input powers and pump detunings on the D1 line, while only excess noise is present on the D2 line. The maximum continuous-wave squeezing observed is -$1.4 \pm0.1$ dB (-2.0 dB corrected for losses). We measure -1.1 dB squeezing at the resonance frequency of the $^{85}$Rb $F=3 \to F'$ transition, which may allow the storage of squeezed light generated by $^{87}$Rb in a $^{85}$Rb quantum memory. Using a pulsed pump, pulsed squeezed light with -1 dB of squeezing for 200 ns pulse widths is observed at 1 MHz repetition rate.Comment: 9 pages, 5 figure

Experimental observation of wave chaos in a conventional optical resonator

Quantum Matter and Optic