ABSTRACT
B cell receptor (BCR) sequencing is a powerful tool for interrogating immune responses to infection and vaccination, but it provides limited information about the antigen specificity of the sequenced BCRs. Here, we present LIBRA-seq (linking B cell receptor to antigen specificity through sequencing), a technology for high-throughput mapping of paired heavy- and light-chain BCR sequences to their cognate antigen specificities. B cells are mixed with a panel of DNA-barcoded antigens so that both the antigen barcode(s) and BCR sequence are recovered via single-cell next-generation sequencing. Using LIBRA-seq, we mapped the antigen specificity of thousands of B cells from two HIV-infected subjects. The predicted specificities were confirmed for a number of HIV- and influenza-specific antibodies, including known and novel broadly neutralizing antibodies. LIBRA-seq will be an integral tool for antibody discovery and vaccine development efforts against a wide range of antigen targets.
Subject(s)
Epitope Mapping/methods , Epitopes/chemistry , Receptors, Antigen, B-Cell/chemistry , Sequence Analysis, DNA/methods , Single-Cell Analysis/methods , Antibodies, Neutralizing/chemistry , Antibodies, Neutralizing/immunology , Antigens/chemistry , Antigens/immunology , Cells, Cultured , Epitopes/immunology , HEK293 Cells , HIV Antibodies/chemistry , HIV Antibodies/immunology , High-Throughput Nucleotide Sequencing/methods , High-Throughput Screening Assays/methods , Humans , Receptors, Antigen, B-Cell/immunology , THP-1 CellsABSTRACT
Diaphragm inactivity during invasive mechanical ventilation leads to diaphragm atrophy and weakness, hemodynamic instability, and ventilatory heterogeneity. Absent respiratory drive and effort can, therefore, worsen injury to both lung and diaphragm and is a major cause of failure to wean. Phrenic nerve stimulation (PNS) can maintain controlled levels of diaphragm activity independent of intrinsic drive and as such may offer a promising approach to achieving lung and diaphragm protective ventilatory targets. Whereas PNS has an established role in the management of chronic respiratory failure, there is emerging interest in how its multisystem putative benefits may be temporarily harnessed in the management of invasively ventilated patients with acute respiratory failure.