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Mitogenig Responses of Sheep Peripheral Blood T-Lymphocytes To Pha and Lps in a Hanging Drop Microassay

Mitogenrespons for T-lymfocyter fra perifert blod hos får for Phytohæmagglutinin (PHA) og lipopolysakkarid (LPS) ved mikroanalyse i hœngende dråbe.

Acta Veterinaria Scandinavica volume 25, pages 548–560 (1984)Cite this article

Abstract

Culture conditions for studying sheep PBL responses to mitogens (PHA and LPS) in a hanging drop micro-assay were investigated. Using those conditions, the responses of putative sheep T-cells (vide infra) to the two mitogens were examined.

The results show that sheep PBL at a cell density of 1–2×106/ml responded optimally to LPS (50 (µg/ml) and PHA (3.125 µg/ml) when cultured in RPMI-1640 in the presence of 2ME, 5% FGS for 48 to 146 h. The cells were cultured as hanging drops (20 µl) in Terasaki plates. Lamb serum failed to support responses to LPS.

T-cells prepared by the nylon wool technique and Helix pomatia-Sepharose-6MB column chromatography responsed vigorously to PHA, but responses to LPS were weak.

Sammendrag

Dyrkningsforhold for studiet af lymfocyter hos får og deres respons til mitogener (PHA og LPS) i hængende dråbe mikroanalyse er undersøgt. Ved denne teknik undersøgtes responsen af formodede T-celler hos får til de to mitogener.

Resultaterne viste, at lymfocyter gav det største respons til LPS (50µg/ml) og PHA (3.126 µg/m1)» når celletætheden var 1–2×106/ ml, og når dyrkning blev foretaget i RPMI-1640 medium tilsat 2-mercaptoethanol og 5 % føtalt kalveserum, over en periode af 48–146 timer. Cellerne blev dyrket på Terasaki skåle i hængende dråbe kulturer (20 µl). Lammeserum kunne ikke benyttes ved LPS stimulation.

T-celler, fremstillet efter nylonvat teknikken og Helix pomatia Sepharose-6MB søjlekromatografi, gav kraftig respons til PHA, mens LPS udvirkede svag response.

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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. Mitogenig Responses of Sheep Peripheral Blood T-Lymphocytes To Pha and Lps in a Hanging Drop Microassay. Acta Vet Scand 25, 548–560 (1984). https://doi.org/10.1186/BF03546922

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Acta Veterinaria Scandinavica

ISSN: 1751-0147