MHC class I presentation and regulation by IFN in bony fish determined by molecular analysis of the class I locus in grass carp.
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.
Chen
Weihong
W
Department of Microbiology and Immunology, College of Veterinary Medicine, China Agricultural University, Beijing, People's Republic of China.
Jia
Zhenghu
Z
Zhang
Ting
T
Zhang
Nianzhi
N
Lin
Changyou
C
Gao
Feng
F
Wang
Li
L
Li
Xiaoying
X
Jiang
Yinan
Y
Li
Xin
X
Gao
George F
GF
Xia
Chun
C
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002347:Carps / Q000235:genetics* / Q000276:immunology; D002460:Cell Line; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D015698:Genomic Library; D015395:Histocompatibility Antigens Class I / Q000145:classification / Q000235:genetics* / Q000276:immunology; D016898:Interferon-alpha / Q000235:genetics* / Q000494:pharmacology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D011110:Polymorphism, Genetic; D011487:Protein Conformation; D017510:Protein Folding; D012209:Rhabdoviridae / Q000187:drug effects; D017422:Sequence Analysis, DNA; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
D029941:Fish Proteins; D015395:Histocompatibility Antigens Class I; D016898:Interferon-alpha