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characterization of the interferon-α response of pigs to the weaning stress

Characterization of the interferon-α response of pigs to the weaning stress.

机构信息

Laboratory of Cellular Immunology, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia-Romagna, Brescia, Italy.

出版信息

J Interferon Cytokine Res. 2011 Feb;31(2):237-47. doi: 10.1089/jir.2010.0041. Epub 2010 Oct 15.

Abstract

The interferon (IFN)-α response of pigs to the stressing event of early weaning was investigated in a field trial. All the animals under study remained healthy and tested negative for common viral infections. However, a low-titered IFN-α response was detected in many sera by a bioassay on Madin-Darby bovine kidney (MDBK) cells on day +6 after weaning. Porcine IFN-α was unambiguously identified by a neutralization assay on a pool of IFN-α-positive sera. By gel filtration chromatography, the antiviral activity of sera on MDBK cells could be traced back to 3 components of apparent molecular mass 27/18/<14 kDa. Additional components of apparent molecular mass 58 and 41 kDa were revealed by ELISA in Nonidet P-40 lysates of peripheral blood mononuclear cells (PBMC). Also, many pigs tested positive in flow cytometry assays on PBMC for intracellular IFN-α. The expression of porcine IFN-α genes was investigated by reverse transcriptase (RT) real-time polymerase chain reaction at days -1, +6, and +12 with regard to weaning in PBMC of 9 piglets. On days -1 and +12, IFN A5, A6, A12, as well as (in fewer pigs) A1, A7, A11, and A2 genes were shown to be expressed. On the contrary, none of the above genes was expressed on day +6, when plenty of pig sera were IFN-α-positive. Our results indicate that weaning causes the release of IFN-α and the transient shut-off of the corresponding gene transcriptions in PBMC. Interestingly, only IFN A9 gene transcription was shown in vitro to be virus induction-dependent.

摘要

本研究通过现场试验,调查了猪在早期断奶应激事件下的干扰素(IFN)-α反应。所有研究动物均保持健康,经常见病毒感染检测呈阴性。然而,在断奶后第 6 天,通过用 Madin-Darby 牛肾(MDBK)细胞进行生物测定,在许多血清中检测到低滴度的 IFN-α反应。通过对一组 IFN-α阳性血清的中和测定,明确鉴定了猪 IFN-α。通过凝胶过滤色谱法,可将 MDBK 细胞血清的抗病毒活性追溯到 3 种表观分子量为 27/18/<14 kDa 的成分。在非离子表面活性剂 P-40 裂解的外周血单核细胞(PBMC)中,通过 ELISA 还发现了表观分子量为 58 和 41 kDa 的其他成分。此外,许多猪在 PBMC 的流式细胞术检测中,细胞内 IFN-α呈阳性。通过逆转录(RT)实时聚合酶链反应,在 PBMC 中调查了 9 头仔猪断奶前后(-1、+6 和+12 天)的猪 IFN-α基因表达。在-1 和+12 天,IFN A5、A6、A12 以及(在较少的猪中)A1、A7、A11 和 A2 基因被证明表达。相反,在大量猪血清 IFN-α呈阳性的+6 天,上述基因均未表达。我们的结果表明,断奶会导致 IFN-α的释放,并使 PBMC 中相应基因转录短暂关闭。有趣的是,仅 IFN A9 基因转录被证明是病毒诱导依赖性的。

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