Investigation of variable lymphocyte receptors in the alternative adaptive immune response of hagfish

• The transcription levels of VLRs, Ikaros and CD45 were time-dependently up-regulated in immunized hagfish.
• The characteristics of the recombinant VLRB proteins varied depending on the number of LRRV modules.
• Production of VLRB in plasma by immunization was both time- and dose-dependent.
• The antigen used for immunization was specifically recognized by VLRB in plasma.

Jawless vertebrates have an alternative adaptive immune system mediated by variable lymphocyte receptors (VLRs), VLRA, VLRC and VLRB. In investigation on the adaptive immunity of hagfish, avian influenza virus hemagglutinin (H9N2-HA1) was used as a model antigen, with mRNA expression levels of VLRA, VLRC and Ikaros were up-regulated in the first week post-immunization. CD45 was up-regulated after the first week; and expression of VLRB progressively increased over the course of the trial. The transcriptional/translational activation of VLRB in blood was verified. The VLRBs cloned from these transcripts showed diversity in their leucine-rich repeats (LRRs). The production of specific VLRB increased in a time- and dose-dependent manner, detected by an anti-VLRB antibody (11G5). The plasma VLRB could distinguish H9N2-HA1 from unrelated proteins, but not from other HA1 subtypes. Together, our findings show that VLRs play a major role in the alternative adaptive immune system of hagfish by responding to specific foreign substances, such as H9N2-HA1.