Validation of a murine proteome-wide phage display library for identification of autoantibody specificities Journal Article
| Authors: | Rackaityte, E.; Proekt, I.; Miller, H. S.; Ramesh, A.; Brooks, J. F.; Kung, A. F.; Mandel-Brehm, C.; Yu, D.; Zamecnik, C. R.; Bair, R.; Vazquez, S. E.; Sunshine, S.; Abram, C. L.; Lowell, C. A.; Rizzuto, G.; Wilson, M. R.; Zikherman, J.; Anderson, M. S.; DeRisi, J. L. |
| Article Title: | Validation of a murine proteome-wide phage display library for identification of autoantibody specificities |
| 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. © 2023, Rackaityte et al. |
| Keywords: | controlled study; unclassified drug; gene mutation; gene sequence; pathogenesis; nonhuman; proteome; mouse; animal; animals; mice; animal tissue; cell protein; validation study; peptide; prediction; molecular cloning; blood sampling; diabetes mellitus; clinical evaluation; peptides; immunoprecipitation; autoantigen; insulin; antibody specificity; autoimmunity; rag2 protein; cross reaction; autoantibody; antibody detection; autoimmune disease; mice, inbred nod; bioinformatics; nonobese diabetic mouse; autoantibodies; phage display; antibody production; adipose tissue; polyendocrinopathy; lupus like syndrome; bacteriophage; bacteriophages; humans; human; male; female; article; genetic profile; pancreatic protein; protein ovalbumin b cell receptor |
| Journal Title: | JCI Insight |
| Volume: | 8 |
| Issue: | 23 |
| ISSN: | 2379-3708 |
| Publisher: | Amer Soc Clinical Investigation Inc |
| Date Published: | 2023-12-08 |
| Start Page: | e174976 |
| Language: | English |
| DOI: | 10.1172/jci.insight.174976 |
| PUBMED: | 37934865 |
| PROVIDER: | scopus |
| PMCID: | PMC10795829 |
| DOI/URL: | |
| Notes: | Article -- Source: Scopus |
