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Antibodies must recognize a great diversity of antigens to protect us from infectious disease. The binding properties of antibodies are determined by the sequences of their corresponding B cell receptors (BCRs). These BCR sequences are created in "draft" form by VDJ recombination, which randomly selects and deletes from the ends of V, D, and J genes, then joins them together with additional random nucleotides. If they pass initial screening and bind an antigen, these sequences then undergo an evolutionary process of mutation and selection, "revising" the BCR to improve binding to its cognate antigen. It has recently become possible to determine the antibody-determining BCR sequences resulting from this process in high throughput. Although these sequences implicitly contain a wealth of information about both antigen exposure and the process by which we learn to resist pathogens, this information can only be extracted using computer algorithms. In this talk, I will describe two recent projects to develop model-based inferential tools for analyzing BCR sequences. In the first, we find that large modern data sets suggest a model using parameter-rich per-allele categorical distributions for HMM transition probabilities and per-allele-per-position mutation probabilities, and that using such a model for VDJ annotation inference leads to significantly improved results. In the second, we investigate selection on BCRs using a novel method that side-steps the difficulties encountered by previous work in differentiating between selection and motif-driven mutation; this is done through stochastic mapping and empirical Bayes estimators that compare the evolution of in-frame and out-of-frame rearrangements. We use this new method to derive a per-residue map of selection on millions of reads, which provides a more nuanced view of the constraints on framework and variable regions.
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Antibodies must recognize a great diversity of antigens to protect us from infectious disease. The binding properties of antibodies are determined by the sequences of their corresponding B cell receptors (BCRs). These BCR sequences are created in "draft" form by VDJ recombination, which randomly selects and deletes from the ends of V, D, and J genes, then joins them together with additional random nucleotides. If they pass initial screening and bind an antigen, these sequences then undergo an evolutionary process of mutation and selection, "revising" the BCR to improve binding to its cognate antigen. It has recently become possible to determine the antibody-determining BCR sequences resulting from this process in high throughput. Although these sequences implicitly contain a wealth of information about both antigen exposure and the process by which we learn to resist pathogens, this information can only be extracted using computer algorithms. In this talk, I will describe two recent projects to develop model-based inferential tools for analyzing BCR sequences. In the first, we find that large modern data sets suggest a model using parameter-rich per-allele categorical distributions for HMM transition probabilities and per-allele-per-position mutation probabilities, and that using such a model for VDJ annotation inference leads to significantly improved results. In the second, we investigate selection on BCRs using a novel method that side-steps the difficulties encountered by previous work in differentiating between selection and motif-driven mutation; this is done through stochastic mapping and empirical Bayes estimators that compare the evolution of in-frame and out-of-frame rearrangements. We use this new method to derive a per-residue map of selection on millions of reads, which provides a more nuanced view of the constraints on framework and variable regions.
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