Regulation of Activation Induced Deaminase (AID) by the hormone estrogen has important implications for understanding adaptive immune responses as well as the involvement of AID in autoimmune diseases and tumorigenesis. This chapter describes the general laboratory techniques for analyzing AID expression and activity induced by estrogen, focusing on the isolation and preparation of cells for hormone treatment and the subsequent analysis of AID responsiveness to estrogen at the RNA level and for determining the regulation of AID activity via estrogen by analyzing Ig switch circle transcripts and mutations in switch region loci.
Methods mol biol
561 - 572
AID, Activation induced deaminase, Adaptive immune response s, Autoimmune disease s, Estrogen, Ig switch circle transcript s, Somatic hypermutation s, Splenic B-cell s, Animals, B-Lymphocytes, Cell Separation, Cytidine Deaminase, Dose-Response Relationship, Drug, Estradiol, Estrogen Antagonists, Estrogens, Female, Gene Expression Regulation, Mice, Inbred BALB C, Mutation, RNA, Messenger, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Spleen, Time Factors, Transcription, Genetic, Workflow