Protein Interactions in Antibody Diversity

National Institute On Aging

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Abstract

Single-cell sequencing of B cell receptors was done to obtain heavy and light chain pairs. We identified and expressed 35 mouse over-represented antibody pairings. Antigen determination was performed by binding the antibodies to proteome arrays and phage immunoprecipitations. 14 mouse protein targets with significant affinity and specificity for the antibodies were selected for further interrogation. Mouse antigens were scrutinized by immunohistochemistry to confirm their presence in inflamed plaques, and these proteins were then expressed to probe autoreactive antibody titers in diseased sera by ELISA. ApoE-deficient mice were immunized with the 7 most promising antigens and were successively placed on a high-fat Western Diet. Serum antibody measurement showed strong, rapid, and early onset of antigen-specific antibody responses in immunized animals, and the mice had increased plaque accumulation in the aortic arch and descending artery.