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

Perifere T-lymfocyter fra fåreblod: Isolering, separering og ouerflade receptorer for Helix pomatia agglutinin (HP) og jordnødde agglutinin (PNA) fra Arachis hypogaea.

Acta Veterinaria Scandinavica volume 25, pages 536–547 (1984)Cite this article

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.

Sammendrag

Undersøgelsen blev gennemført med henblik på at udvikle og evaluere teknikker til isolering af ovine, perifere blodlymfocyter (PBL) og med henblik på at undersøge egnetheden af jornødde agglutinin (peanut agglutinin, PNA) og Helix pomatia agglutinin (HP) som markør for fare T celler.

Resultaterne viser, at ovine PBL kan prsepareres reproducerbart ved at inkubere färeblod med carbonyl jern, centrifugere i Percoll (kolloidt silicium polyvinylpyrrolidin) separeringsmedium og derefter at høste PBL fra interfasen. PBL, der er præpareret efter denne metode, agglutinerer sjældent spontant, hvilket ellers ofte ses med buffy coat celler og perifere blod mononukleœre celler.

PNA og HP kan benyttes som ovine T celle markører, fordi disse lektiner under egnede betingelser ikke bindes til B celler. Højt oprensede perifere blod-lymfocyter kunne med godt resultat præpareres ved hjælp af nylon uld teknikken og affinitetssøjle kromatografi med HP, men derimod ikke med HP-Sepharose-6MB kromatografi søjler.

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  1. Department of Veterinary Virology and Immunology, Royal Veterinary and Agricultural University, Copenhagen, Denmark

    J. E. D. Mlangwa

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Mlangwa, J.E.D. Sheep Peripheral Blood-T-Lymphocytes: Isolation, Separation and Surface Receptors for Helix Pomatia Agglutinin and Peanut Agglutinin From Arachis Hypogaea. Acta Vet Scand 25, 536–547 (1984). https://doi.org/10.1186/BF03546921

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Acta Veterinaria Scandinavica

ISSN: 1751-0147