학술논문
Validation of a murine proteome-wide phage display library for identification of autoantibody specificities
Document Type
article
Author
Rackaityte, Elze; Proekt, Irina; Miller, Haleigh S; Ramesh, Akshaya; Brooks, Jeremy F; Kung, Andrew F; Mandel-Brehm, Caleigh; Yu, David JL; Zamecnik, Colin R; Bair, Rebecca; Vazquez, Sara E; Sunshine, Sara; Abram, Clare L; Lowell, Clifford A; Rizzuto, Gabrielle; Wilson, Michael R; Zikherman, Julie; Anderson, Mark S; DeRisi, Joseph L
Source
JCI Insight. 8(23)
Subject
Language
Abstract
Autoimmunity is characterized by loss of tolerance to tissue-specific as well as systemic antigens, resulting in complex autoantibody landscapes. Here, we introduce and extensively validate the performance characteristics of a murine proteome-wide library for phage display immunoprecipitation and sequencing (PhIP-seq) in profiling mouse autoantibodies. This library was validated using 7 genetically distinct mouse lines across a spectrum of autoreactivity. Mice deficient in antibody production (Rag2-/- and μMT) were used to model nonspecific peptide enrichments, while cross-reactivity was evaluated using anti-ovalbumin B cell receptor-restricted OB1 mice as a proof of principle. The PhIP-seq approach was then utilized to interrogate 3 distinct autoimmune disease models. First, serum from Lyn-/- IgD+/- mice with lupus-like disease was used to identify nuclear and apoptotic bleb reactivities. Second, serum from nonobese diabetic (NOD) mice, a polygenic model of pancreas-specific autoimmunity, was enriched in peptides derived from both insulin and predicted pancreatic proteins. Lastly, Aire-/- mouse sera were used to identify numerous autoantigens, many of which were also observed in previous studies of humans with autoimmune polyendocrinopathy syndrome type 1 carrying recessive mutations in AIRE. These experiments support the use of murine proteome-wide PhIP-seq for antigenic profiling and autoantibody discovery, which may be employed to study a range of immune perturbations in mouse models of autoimmunity profiling.