1D-IEF analysis of BoLA class I expression using allo-antisera reveals additional complexity.
The use of the bovine allo-antisera in lymphocyte microcytotoxicity assays suggests that there is a single highly polymorphic class I product expressed by the BoLA system encoded by one locus. In contrast, biochemical techniques, such as 1D-IEF, reveal a complex pattern of bands for BoLA class I molecules from each animal. In order to understand the origins of this heterogeneity bovine allo-antisera were used in the immunoprecipitation step of 1D-IEF and the results compared with those from immunoprecipitation using the monoclonal antibody W6/32. By modifying existing protocols to include Gammabind G a range of bovine allo-antisera were used successfully to immunoprecipitate bovine MHC class I molecules. The results indicate that the bovine allo-antisera do not recognize all molecules previously assigned to BoLA class I serotypes by 1D-IEF. Furthermore, some of the allo-antisera immunoprecipitated molecules are not recognized by W6/32 and vice versa. This suggests that more than one polymorphic locus is expressed from the bovine MHC and that each allo-antiserum recognizes molecules encoded by different loci. Examination of the results also suggests the existence of linkage disequilibrium in the BoLA class I region.
al-Murrani
S W
SW
AFRC Roslin Institute (Edinburgh), Roslin, Midlothian, UK.
Glass
E J
EJ
Williams
J L
JL
Oliver
R A
RA
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions / Q000235:genetics; D002417:Cattle / Q000235:genetics* / Q000276:immunology*; D004247:DNA / Q000032:analysis; D015870:Gene Expression; D005805:Genes, MHC Class I / Q000235:genetics*; D006239:Haplotypes; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D007518:Isoantibodies / Q000235:genetics / Q000276:immunology*; D007525:Isoelectric Focusing / Q000662:veterinary; D015810:Linkage Disequilibrium; D008214:Lymphocytes; D011233:Precipitin Tests / Q000662:veterinary
D000911:Antibodies, Monoclonal; D015395:Histocompatibility Antigens Class I; D007106:Immune Sera; D007518:Isoantibodies; D004247:DNA