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Primary Structure and Complementarity-Determining Region (CDR) 3 Spectratyping of Rainbow Trout TCR Transcripts Identify Ten V Families with V6 Displaying Unusual CDR2 and Differently Spliced Forms¹

Institut National de la Recherche Agronomique, Unité de Virologie et Immunologie Moléculaires, Jouy-en-Josas, France

VDJ rearrangement at the teleost TCR locus leads to a highly diverse repertoire of junctions for each VJ combination. From a rainbow trout 5' RACE library of TCR transcripts, 47 clones encompassing a full V-D-J-C sequence were selected and analyzed. A similarity analysis of the sequences evidenced 10 V families, of which 6 were not previously described. Immunoscope and sequence analysis of the V-D-J junctions of the new families confirmed that they create a polyclonal and diverse repertoire. Multiple alignments showed that rainbow trout Vs possess most of the conserved residues typical of V segments. However, this study revealed a high complementarity-determining region 2 (CDR2) and CDR1 length diversity among rainbow trout V families, suggesting that the spatial orientation of the TCR could fluctuate in the TCR/peptide/MHC complex, depending on the V expressed. Among the new V families, V6 displayed the strongest deviance from typical hypervariable CDR1 and CDR2 loops, with an unusually short CDR2. Moreover, the V6 sequence is overall divergent from typical V sequence, raising the question of its functional relevance. Immunoscope experiments identified a V6-J3 junction, which was amplified during the response against viral hemorrhagic septicemia virus, a fish rhabdovirus. V6 seems therefore to be expressed functionally in a selected TCR. However, the shorter V6 transcripts produced through an alternative splicing lack the C', C'', D, and E strands of the V domain and are probably nonfunctional.

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