Nylon wool column fractionation and characterisation of feline lymphocyte subpopulations

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Abstract

Feline separated mononuclear cells (SMC) were obtained from peripheral blood by ficoll-diatrizoate gradient separation. SMC were further fractionated on nlyon wool columns into nylon wool adherent cells (NWAC) and nylon wool effluent cells (NWEC). The three cell populations, SMC, NWAC and NWEC, were characterised using direct immunofluorescent staining for surface immunoglobulin (sIg) as a B cell marker and neuramidase treated guinea pig erythrocyte-rosette formation (E-rosettes) and mitogen-induced lymphocye blastogenesis (LB) as possible T-cell markers.

Feline SMC consisted of 30.1 ± 4.0% sIg+ cells 36.6 + 5.4% E-rosette forming cells and 33.3% null cells i.e. cells which were sIg and non E-rosette forming. Fractionation of SMC on nylon wool columns yielded NWEC which were significantly enriched for T cells in that they contained 68.6 ± 2.9% E-rosette forming cells. NWAC were 51.0% ± 10.8% sIg+, ∼ 20% of cells were lost. The LB responsiveness of NWEC to concanavalin A (Con A) and phytonaemagglutinin-P (PHA-P) was enhanced compared to SMC. NWAC were non-responsive to Con A and PHA-P at all concentrations tested.

It was concluded that nylon wool column fractionation of feline SMC was an efficient procedure for T cell enrichment and that the enriched cells retained the properties of E-rosette formation and blastogenesis by mitogens.

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