Molecular and functional profiling of antigen-driven B-cell responses at single cell resolution in autoimmunity and infectious disease
Abstract/Contents
- Abstract
- Innovations in deep sequencing have enabled the analysis of single cell transcriptomes and immune repertoires in immune cell populations. Despite this, the mechanisms underlying humoral responses in many autoimmune and infectious diseases contexts remain poorly understood due to technical limitations that prevent the characterization of relationships between immune receptor and transcriptome as well as functional analyses of prominent antibodies. Here, we develop and utilize next-generation sequencing approaches to understand B-cell responses in patients with rheumatoid arthritis and in patients with Staphylococcus aureus infection. First, we utilize a novel single-cell RNA-seq approach that enables simultaneous transcriptomic profiling and B-cell receptor (BCR) reconstitution to analyze subset-specific dysregulation in tetramer-positive B cells from RA patients that target RF (RF) and citrullinated proteins (ACPA). We find that B cells in naive, memory, and plasmablast stages all target RF and ACPA, indicating mechanisms of defective central and peripheral tolerance. We also elucidate affinity maturation gene signatures, innate immune signatures, and coexpression pathways that differentiate the RF and ACPA responses, demonstrating that unique costimulatory pathways are required for different autoantigen responses. Second, we perform deep sequencing of paired IgG plasmablast antibody repertoires in Staphylococcus aureus bacteremia patients who eventually recovered from infection to identify rapidly expanding B cell clones during infection and enable rational selection of antibodies for recombinant expression. Through functional characterization of antibodies from S. aureus patients, we identify fibronectin binding protein A as a prominent antigen in productive humoral responses and show that recombinant antibodies promote opsonophagocytosis. Taken together, these studies demonstrate the ability of single cell B-cell analyses to uncover the functional heterogeneity of the humoral response, characterize transcriptional profiles at different stages of B-cell differentiation, and identify antigens targeted by the active antibody response.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2017 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Lu, Daniel Ryan | |
|---|---|---|
| Associated with | Stanford University, Department of Immunology. | |
| Primary advisor | Robinson, William (William Hewitt) | |
| Thesis advisor | Robinson, William (William Hewitt) | |
| Thesis advisor | Amieva, Manuel | |
| Thesis advisor | Fathman, C. Garrison | |
| Thesis advisor | Goronzy, J. J | |
| Advisor | Amieva, Manuel | |
| Advisor | Fathman, C. Garrison | |
| Advisor | Goronzy, J. J |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Daniel Ryan Lu. |
|---|---|
| Note | Submitted to the Department of Immunology. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2017. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2017 by Daniel Ryan Lu
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