Sheep Peripheral Blood-T-Lymphocytes: Isolation, Separation and Surface Receptors for Helix Pomatia Agglutinin and Peanut Agglutinin From Arachis Hypogaea

Abstract

A study was made to develop and evaluate techniques for the isolation of ovine peripheral blood lymphocytes (PBL) and to investigate the suitability of peanut agglutinin (PNA) and Helix pomatia agglutinin (HP) as markers for sheep T-cells. The results show that sheen PBL can be prepared reproducibly by incubating ovine blood with carbonyl iron, centrifuging the blood over Percoll (colloidal silica polyvinylpyrrolidine) separating media (Pharmacia, Sweden) and harvesting the PBL at the interface. PBL so prepared, rarely undergo spontaneous agglutination, which is frequently seen with buffy coat cells and peripheral blood mononuclear cells. PNA and HP can be used as markers for ovine T-lymphocytes, because, under appropriate conditions, these lectins do not bind to B cells. Highly enriched peripheral blood T-lymphocytes were successfully prepared by the nylon wool technique and affinity column chromatography using HP. Highly purified B-cell sub-populations could not be prepared using the HP-Sepharose-6MB chromatography columns.