Chicken major histocompatibility complex class I definition using antisera induced by cloned class I sequences.
Alloantisera directed against chicken class I MHC (BFIV) antigens were produced by using transfected cell lines expressing cloned BFIV sequences. The cloned BFIV sequences were from haplotypes *12, *13, and *21. Two laboratory-derived class I mutant sequences (BFIV13m126 and BFIV21m78) were developed to analyze cross-reactive epitopes and to induce specific alloantisera. Antisera were tested in hemagglutination and flow cytometry assays. The antisera produced were highly specific and had minimal cross-reactivity. The antisera induced by the BF1V21m78 mutant confirmed the significance of amino acids 78 and 81 in cross-reactivity between haplotypes B*21 and B*5. The highly specific antisera were tested by hemagglutination on red blood cells of 31 different MHC haplotypes. The consistency of hemagglutination patterns and minimal cross-reactivity demonstrated that these BFIV antisera are extremely valuable in defining MHC haplotype in various chicken lines. Because of the extreme low level of recombination between the chicken class I and class II loci, identification of BFIV allele can be used to define MHC haplotype within a line. Complete identity between the transfected cell line and the chicken used to produce the antiserum is required to ensure the monospecificity.
Fulton
J E
JE
USDA/ARS, Avian Disease and Oncology Lab, East Lansing, Michigan 48823, USA.
Hunt
H D
HD
Bacon
L D
LD
D000483:Alleles; D000818:Animals; D000918:Antibody Specificity; D002460:Cell Line; D002645:Chickens / Q000276:immunology*; D003001:Cloning, Molecular; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D005434:Flow Cytometry; D006239:Haplotypes; D006384:Hemagglutination; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology*; D016296:Mutagenesis; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D014162:Transfection
D000939:Epitopes; D015395:Histocompatibility Antigens Class I; D007106:Immune Sera; D010446:Peptide Fragments