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- W2077463014 abstract "Gamma interferon produced by porcine lymphocytes (nPoIFNγ) in response to stimulation with phorbol-myristate-acetate (PMA) and phytohemagglutinin (PHA) was monitored by a radioimmunoassay (RIA). The RIA was developed with antibodies raised in rabbits against recombinant porcine IFNγ (rPoIFNγ) and made monospecific on a rPoIFNγ/Sepharose matrix. This anti-PoIFNγ antibody was shown to bind and neutralize rPoIFNγ specifically and to cross-react with bovine IFNγ but not with murine IFNγ or porcine IFNα. The nPoIFNγ levels produced by lymphocytes in response to PMA and PHA were at least two-fold higher than control lymphocytes as measured by the RIA in culture fluids. These culture fluids were fractionated on Concanavalin A Sepharose (Con A/Seph) and anti-rPoIFNγ/Seph in attempts to evaluate the induced nPoIFNγ further. The separation was monitored by RIA and showed that nPoIFNγ was retained on Con A/Seph suggesting the presence of sugar residues on the molecules. Pools of Con A/Seph fractions, positive in RIA, were separated further on the anti-rPoIFNγ/Seph matrix where a total adsorption of nPoIFNγ occurred. On a weight basis, the eluates from the anti-rPoIFNγ/Seph had a reactivity in RIA at least four times higher than the fractions derived from Con A/Seph. This indicated that the nPoIFNγ remained immunochemically reactive after being eluted from Con A/Seph and that the separation on anti-rPoIFNγ/Seph chromatography was specific. Purified nPoIFNγ exhibited a major band with the same migration characteristics of rPoIFNγ in PAGE-SDS electrophoresis and several minor bands when reacted with 125I anti-rPoIFNγ antibody in Western blots. A total loss of reactivity to antibody after radioiodination of nPoIFNγ, however, prevented the confirmation of these results by immunoprecipitation. A direct, rapid and sensitive immunoassay for measurement of the relative levels of nPoIFNγ present in biological fluids is presented and this method could be a useful complement to bioassays for IFNγ." @default.
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- W2077463014 date "1994-03-01" @default.
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- W2077463014 title "Porcine lymphocyte gamma interferon responses to mitogenic stimuli monitored by a direct immunoassay" @default.
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- W2077463014 doi "https://doi.org/10.1016/0165-2427(94)90020-5" @default.
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