Molecular and cellular analysis of B-cell populations in the rainbow trout using Pax5 and immunoglobulin markers

To date, the trout B-cell is poorly defined, as many essential molecular markers are not yet available for this species. In mammalian systems, the transcription factor Pax5, expressed from pre-B through plasmablast stages, provides an important marker for B-cell differentiation. In a previous study we showed that Pax5 is expressed in the trout. Here we identify trout B-cell populations that vary in expression of Pax5, membrane and secreted Ig. Immune tissues were separated based on concentration of surface IgM, and analyzed by qPCR and flow cytometry. Results suggest that spleen and PBL contain mostly resting B cells, which lack secreted Ig. While the great majority of splenic B cells become strongly activated upon LPS stimulation, PBLs do not. Additionally, anterior kidney contains both developing B and Ig-secreting B-cell populations, but few resting, mature B cells. Lastly, posterior kidney contains multiple B-cell populations in various states of activation. We conclude that trout immune tissues contain multiple, developmentally diverse and tissue-specific B-cell populations as defined by their relative expression of Pax5, surface IgM, and secreted IgM.