FcγRIIb differentially regulates pre-immune and germinal center B cell tolerance in mouse and human.

Several tolerance “checkpoints” exist throughout B cell development to control autoreactive B cells and prevent the generation of pathogenic autoantibodies. FcγRIIb is an Fc receptor that inhibits B cell activation and, if defective, is associated with autoimmune disease. Its impact on specific B cell tolerance checkpoints is unknown. Here we show that reduced expression of FcγRIIb leads to increased deletion and anergy of autoreactive immature B cells, but despite this autoreactive B cells expand in the germinal center and serum autoantibodies are produced, even in response to exogenous non-self antigen. Thus, we show FcγRIIb has opposing effects on pre- and post-immune tolerance checkpoints, and suggest B cell tolerance requires the control of “bystander” germinal center B cells with low or no affinity for the immunization antigen.This work was funded by the Wellcome Trust (Programme Grant Number 083650/Z/07/Z to KGCS) and supported by the NIHR Cambridge Biomedical Research Centre. ME was funded by the Wellcome Trust (Programme Grant Number 083650/Z/07/Z), by a Junior Team Leader starting grant from the Laboratory of Excellence in Research on Medication and Innovative Therapeutics (LabEx LERMIT) supported by a grant from ANR (ANR-10-LABX-33) under the program “Investissements d'Avenir” (ANR-11-IDEX-0003-01) and by an ANR @RAction starting grant (ANR-14-ACHN- 0008). KGCS is an NIHR Senior Clinical Investigator and a Distinguished Innovator of the Lupus Research Institute

Dynamics of immunoglobulin sequence diversity in HIV-1 infected individuals.

Advances in immunoglobulin (Ig) sequencing technology are leading to new perspectives on immune system dynamics. Much research in this nascent field has focused on resolving immune responses to viral infection. However, the dynamics of B-cell diversity in early HIV infection, and in response to anti-retroviral therapy, are still poorly understood. Here, we investigate these dynamics through bulk Ig sequencing of samples collected over 2 years from a group of eight HIV-1 infected patients, five of whom received anti-retroviral therapy during the first half of the study period. We applied previously published methods for visualizing and quantifying B-cell sequence diversity, including the Gini index, and compared their efficacy to alternative measures. While we found significantly greater clonal structure in HIV-infected patients versus healthy controls, within HIV patients, we observed no significant relationships between statistics of B-cell clonal expansion and clinical variables such as viral load and CD4+ count. Although there are many potential explanations for this, we suggest that important factors include poor sampling resolution and complex B-cell dynamics that are difficult to summarize using simple summary statistics. Importantly, we find a significant association between observed Gini indices and sequencing read depth, and we conclude that more robust analytical methods and a closer integration of experimental and theoretical work is needed to further our understanding of B-cell repertoire diversity during viral infection

Use of prescribed and non-prescribed medication for dyspepsia.

Contains fulltext : 59130.pdf (publisher's version ) (Open Access)OBJECTIVE: To explore patient factors related to the use of prescribed and non-prescribed drugs for dyspepsia in The Netherlands. DESIGN: Patient survey study. SETTING AND SUBJECTS: Questionnaires sent to patients who had a prescription for dyspepsia medication from their general practitioner. MAIN OUTCOME MEASURES: Patient factors related to the on-demand use of prescribed medication and the use of non-prescribed medication for dyspepsia. RESULTS: 74% of the (n=518) patients had been receiving prescribed medication for dyspepsia for more than one year. A quarter of the patients were using the prescribed medication "on demand" instead of adhering to the instructions on the prescription. PPI prescriptions reduced the probability of using the medication on demand, compared with other prescribed drugs (OR 0.39). Some 19% of the patients were using non-prescribed drugs for dyspepsia. More of the patients who had visited their general practitioner in the previous 12 months were using their drugs on demand (OR 2.27) and were using non-prescribed drugs (OR 2.40) than the patients who had not visited their GP. CONCLUSION: Clear information for patients on how to use their medication for dyspepsia may contribute to decreasing unnecessary drug use. Communication about (in)appropriate use of drugs "on demand", non-prescribed drugs, and health education should be addressed to all patient groups. Further studies on these topics should aim to improve medical care based on shared decision-making for patients with dyspepsia

Antigen Receptor Sequence Reconstruction and Clonality Inference from scRNA-Seq Data

In this chapter, we describe TraCeR and BraCeR, our computational tools for reconstruction of paired full-length antigen receptor sequences and clonality inference from single-cell RNA-seq (scRNA-seq) data. In brief, TraCeR reconstructs T-cell receptor (TCR) sequences from scRNA-seq data by extracting sequencing reads derived from TCRs by aligning the reads from each cell against synthetic TCR sequences. TCR-derived reads are then assembled into full-length recombined TCR sequences. BraCeR builds on the TraCeR pipeline and accounts for somatic hypermutations (SHM) and isotype switching. Here we discuss experimental design, use of the tools, and interpretation of the results