MHC Class I Presentation and Regulation by IFN in Bony Fish Determined by Molecular Analysis of the Class I Locus in Grass Carp

Abstract

Beyond their sequences, little is known regarding MHC class I presentation and regulation by IFN in bony fish. In this work, the class I locus (Ctid-UBA) was isolated from a grass carp fosmid library, and its polymorphisms and tissue expression were investigated. The Ctid-UBA and Ctid-beta2-microglobulin genes then were expressed and refolded, and tetramer techniques were used to identify the CTL response. The interaction between grass carp type I IFN and Ctid-UBA genes was investigated. Two fosmids coding for Ctid-UBA *0101 and Ctid-UBA *0201 genes were sequenced. The SXY box and IFN-stimulated regulatory element motifs were located from the start codons to -800 bp in Ctid-UBA. A Southern blot showed three to four bands, suggesting that grass carp contains at least three class I loci. In addition, the Ctid-UBA allelic genes are expressed in all tissue of grass carp. The three-dimensional structure of Ctid-UBA *0102 showed that the peptide-binding domain was formed by the alpha1 and alpha2 domains, which could bind several nonapeptides of grass carp hemorrhagic virus. There were 1.60% more PE-positive cells in P1(QPNEAIRSL)-immunized fish than in blank and adjuvant control groups. Additionally, recombinant grass carp IFN could regulate the expression of Ctid-UBA. These results characterize the class I presentation, CTL response, and regulation by type I IFN in bony fish.