Discovery of a unique Ig heavy-chain isotype (IgT) in rainbow trout: Implications for a distinctive B cell developmental pathway in teleost fish.
During the analysis of Ig superfamily members within the available rainbow trout (Oncorhynchus mykiss) EST gene index, we identified a unique Ig heavy-chain (IgH) isotype. cDNAs encoding this isotype are composed of a typical IgH leader sequence and a VDJ rearranged segment followed by four Ig superfamily C-1 domains represented as either membrane-bound or secretory versions. Because teleost fish were previously thought to encode and express only two IgH isotypes (IgM and IgD) for their humoral immune repertoire, we isolated all three cDNA isotypes from a single homozygous trout (OSU-142) to confirm that all three are indeed independent isotypes. Bioinformatic and phylogenetic analysis indicates that this previously undescribed divergent isotype is restricted to bony fish, thus we have named this isotype "IgT" (tau) for teleost fish. Genomic sequence analysis of an OSU-142 bacterial artificial chromosome (BAC) clone positive for all three IgH isotypes revealed that IgT utilizes the standard rainbow trout V(H) families, but surprisingly, the IgT isotype possesses its own exclusive set of D(H) and J(H) elements for the generation of diversity. The IgT D and J segments and tau constant (C) region genes are located upstream of the D and J elements for IgM, representing a genomic IgH architecture that has not been observed in any other vertebrate class. All three isotypes are primarily expressed in the spleen and pronephros (bone marrow equivalent), and ontogenically, expression of IgT is present 4 d before hatching in developing embryos.
Hansen
John D
JD
Western Fisheries Research Center, Biological Resources Division and Department of Pathobiology, U.S. Geological Survey and University of Washington, Seattle, WA 98115, USA. jdh25@u.washington.edu
Landis
Eric D
ED
Phillips
Ruth B
RB
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D001853:Bone Marrow / Q000378:metabolism; D022202:Chromosomes, Artificial, Bacterial; D019295:Computational Biology / Q000379:methods; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D020224:Expressed Sequence Tags; D005803:Genes, Immunoglobulin / Q000235:genetics; D006720:Homozygote; D007072:Immunoglobulin D / Q000737:chemistry; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry*; D007075:Immunoglobulin M / Q000737:chemistry; D008957:Models, Genetic; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss; D010802:Phylogeny; D020161:Physical Chromosome Mapping; D017434:Protein Structure, Tertiary; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045567:VDJ Recombinases / Q000235:genetics; D015027:Zebrafish
D017931:DNA Primers; D018076:DNA, Complementary; D007072:Immunoglobulin D; D007143:Immunoglobulin Heavy Chains; D007075:Immunoglobulin M; D045567:VDJ Recombinases