RESUMO
A method is described for the purification of sheep lymphocytes carrying class II MHC antigens. After incubation of purified blood lymphocytes on anti-IgM-coated petri dishes, the adherent fraction contained 95% sIg-positive cells determined by immunofluorescence. When tested with cross-reacting anti-class II (bovine and human) monoclonal antibodies, more than 95% of these cells were positive either by immunofluorescence or cytotoxicity. This technique will permit studies of the polymorphism of sheep class II antigens.
Assuntos
Linfócitos B/imunologia , Antígenos de Histocompatibilidade/análise , Leucócitos/imunologia , Receptores de Antígenos de Linfócitos B/análise , Ovinos/imunologia , Animais , Anticorpos Monoclonais , Separação Celular/métodos , Cabras/imunologia , Complexo Principal de Histocompatibilidade , Especificidade da Espécie , Suínos/imunologiaRESUMO
A horse family in which a recombination occurred in the chromosome region coding for the serological specificities of the ELA complex and those of the A blood group system of a mare was further analysed by mixed lymphocyte reaction (MLR) and Southern blot hybridization. This family consisted of a stallion, a mare and five full sibs. The stallion and the mare were heterozygous for internationally recognized ELA specificities while only the mare was heterozygous for the A blood group system. MLR between all members of the family confirmed that the stallion possessed two different ELA haplotypes and suggested that recombination in the mare occurred outside the segment delimited by the ELA-A locus and the MLR region. DNA samples from all individuals were investigated by Southern blot analysis using three restriction enzymes (EcoRI, HindIII or TaqI), three human HLA probes (one of class I cDNA and two of class II probes), one cDNA (DR beta) and one genomic (DQ alpha). Class I and class II restriction fragments of the mare segregated in accordance to the ELA specificities and thus clearly confirming that the crossing-over did not occur between the ELA-A gene and the class I, class II region nor between DR beta and DQ alpha subsets. The A blood group genetic determinants would thus be situated outside the ELA region defined by class I and class II genes.