In Vitro Evolution of Antibodies Inspired by In Vivo Evolution

In vitro generation of antibodies often requires variable domain sequence evolution to adapt the protein in terms of affinity, specificity, or developability. Such antibodies, including those that are of interest for clinical development, may have their origins in a diversity of immunoglobulin germline genes. Others and we have previously shown that antibodies of different origins tend to evolve along different, preferred trajectories. Apart from substitutions within the complementary determining regions, evolution may also, in a germline gene-origin-defined manner, be focused to residues in the framework regions, and even to residues within the protein core, in many instances at a substantial distance from the antibody’s antigen-binding site. Examples of such germline origin-defined patterns of evolution are described. We propose that germline gene-preferred substitution patterns offer attractive alternatives that should be considered in efforts to evolve antibodies intended for therapeutic use with respect to appropriate affinity, specificity, and product developability. We also hypothesize that such germline gene-origin-defined in vitro evolution hold potential to result in products with limited immunogenicity, as similarly evolved antibodies will be parts of conventional, in vivo-generated antibody responses and thus are likely to have been seen by the immune system in the past

Women In the weighing room: gender discrimination on the thoroughbred racetrack

Women jockeys are a small minority on the thoroughbred racetrack and securesignificantly fewer racing mounts than their male counterparts. This suggests female jockeys are facing discriminatory barriers, in one of the only major professional sportswhere men and women compete against each other on equal terms. This exploratorystudy considers discriminatory barriers that exist and the effects they have on women’scomparative profile and participation in the flat racing industry. Six participants wererecruited for the study from different areas within the industry, and with at least threeyears experience. Information was derived from semi structured individual interviews. The data was analysed using discourse analysis techniques. Five main themes wereevident: a culture of sexism, including the sense that women are more nurturing;opportunities, including for women to become trainers; body shape and strength; riskand danger; industry fashion and trends. The results from this study suggest women face discrimination in horseracing onaccount of a number of factors, the three main perceived reasons are due to theirphysical strength, body shape and the tradition and history embedded within theindustry. Whilst there is a shift starting to occur where more women are coming throughin flat racing, this is slow. Participants consider that women may find these barriers andperceptions held by others difficult to overcome, which may result in their inability toachieve equality in this sport. Given the exploratory character of the study, conclusionsare tentative and we propose a number of areas for further research

Single-cell BCR and transcriptome analysis after influenza infection reveals spatiotemporal dynamics of antigen-specific B cells

B cell responses are critical for antiviral immunity. However, a comprehensive picture of antigen-specific B cell differentiation, clonal proliferation, and dynamics in different organs after infection is lacking. Here, by combining single-cell RNA and B cell receptor (BCR) sequencing of antigen-specific cells in lymph nodes, spleen, and lungs after influenza infection in mice, we identify several germinal center (GC) B cell subpopulations and organ-specific differences that persist over the course of the response. We discover transcriptional differences between memory cells in lungs and lymphoid organs and organ-restricted clonal expansion. Remarkably, we find significant clonal overlap between GC-derived memory and plasma cells. By combining BCR-mutational analyses with monoclonal antibody (mAb) expression and affinity measurements, we find that memory B cells are highly diverse and can be selected from both low- and high-affinity precursors. By linking antigen recognition with transcriptional programming, clonal proliferation, and differentiation, these finding provide important advances in our understanding of antiviral immunity

Inferred Allelic Variants of Immunoglobulin Receptor Genes: a system for their evaluation, documentation, and naming

Immunoglobulins or antibodies are the main effector molecules of the B-cell lineage and are encoded by hundreds of variable (V), diversity (D), and joining (J) germline genes, which recombine to generate enormous IG diversity. Recently, high-throughput adaptive immune receptor repertoire sequencing (AIRR-seq) of recombined V-(D)-J genes has offered unprecedented insights into the dynamics of IG repertoires in health and disease. Faithful biological interpretation of AIRR-seq studies depends upon the annotation of raw AIRR-seq data, using reference germline gene databases to identify the germline genes within each rearrangement. Existing reference databases are incomplete, as shown by recent AIRR-seq studies that have inferred the existence of many previously unreported polymorphisms. Completing the documentation of genetic variation in germline gene databases is therefore of crucial importance. Lymphocyte receptor genes and alleles are currently assigned by the Immunoglobulins, T cell Receptors and Major Histocompatibility Nomenclature Subcommittee of the International Union of Immunological Societies (IUIS) and managed in IMGT®, the international ImMunoGeneTics information system® (IMGT). In 2017, the IMGT Group reached agreement with a group of AIRR-seq researchers on the principles of a streamlined process for identifying and naming inferred allelic sequences, for their incorporation into IMGT®. These researchers represented the AIRR Community, a network of over 300 researchers whose objective is to promote all aspects of immunoglobulin and T-cell receptor repertoire studies, including the standardization of experimental and computational aspects of AIRR-seq data generation and analysis. The Inferred Allele Review Committee (IARC) was established by the AIRR Community to devise policies, criteria, and procedures to perform this function. Formalized evaluations of novel inferred sequences have now begun and submissions are invited via a new dedicated portal (https://ogrdb.airr-community.org). Here, we summarize recommendations developed by the IARC—focusing, to begin with, on human IGHV genes—with the goal of facilitating the acceptance of inferred allelic variants of germline IGHV genes. We believe that this initiative will improve the quality of AIRR-seq studies by facilitating the description of human IG germline gene variation, and that in time, it will expand to the documentation of TR and IG genes in many vertebrate species

Rocken som murbräcka

Hur mådde rock'n'rollen i DDR? Bättre än man kan tro, menar Mats Greiff. Genom den underjordiska musikkulturen lärde sig flera generationer östtyskar hur de skulle organisera sig, samarbeta och föra statens kontrollanter. Rockmusiken var läroprocessen som förberedde östtyskarna för den fredliga revolutionen

Hästkapplöpningar och kapitalismens framväxt i Storbritannien

Recension av Mike Huggins, ”Horse Racing and British Society in the Long Eighteenth Century