Transcription analysis of two Eomesodermin genes in lymphocyte subsets of two teleost species

• Two Eomes genes Eomes-a and -b were cloned from rainbow trout and ginbuna.
• Both Eomes transcripts were detected in lymphoid organs.
• Pronephric CD8α+ lymphocytes expressed both Eomes transcripts.
• Rainbow trout Eomes were differently expressed between non-mucosal and mucosal CD8α+ lymphocytes.
• Rainbow trout CD8α+ mucosal lymphocytes displayed high amounts of IFN-γ transcripts.

Eomesodermin (Eomes), a T-box transcription factor, is a key molecule associated with function and differentiation of CD8+ T cells and NK cells. Previously, two teleost Eomes genes (Eomes-a and -b), which are located on different chromosomes, were identified and shown to be expressed in zebrafish lymphocytes. For the present study, we identified these genes in rainbow trout and ginbuna crucian carp. Deduced Eomes-a and -b amino acid sequences in both fish species contain a highly conserved T-box DNA binding domain. In RT-PCR, both Eomes transcripts were readily detectable in a variety of tissues in rainbow trout and ginbuna. The high expression of Eomes-a and -b in brain and ovary suggests involvement in neurogenesis and oogenesis, respectively, while their expression in lymphoid tissues presumably is associated with immune functions. Investigation of separated lymphocyte populations from pronephros indicated that both Eomes-a and -b transcripts were few or absent in IgM+ lymphocytes, while relatively abundant in IgM−/CD8α+ and IgM−/CD8α− populations. Moreover, we sorted trout CD8α+ lymphocytes from mucosal and non-mucosal lymphoid tissues and compared the expression profiles of Eomes-a and -b with those of other T cell-related transcription factor genes (GATA-3, T-bet and Runx3), a Th1 cytokine gene (IFN-γ) and a Th2 cytokine gene (IL-4/13A). Interestingly, the tissue distribution of Eomes-a/b, T-bet, and Runx3 versus IFN-γ transcripts did not reveal simple correlations, suggesting tissue-specific properties of CD8α+ lymphocytes and/or multiple modes that drive IFN-γ expressions.