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猪初乳和外周血中T细胞基因表达谱的比较。

Comparison of gene expression profiles of T cells in porcine colostrum and peripheral blood.

作者信息

Ogawa Shohei, Okutani Mie, Tsukahara Takamitsu, Nakanishi Nobuo, Kato Yoshihiro, Fukuta Kikuto, Romero-Pérez Gustavo A, Ushida Kazunari, Inoue Ryo

出版信息

Am J Vet Res. 2016 Sep;77(9):961-8. doi: 10.2460/ajvr.77.9.961.

Abstract

OBJECTIVE To compare gene expression patterns of T cells in porcine colostrum and peripheral blood. ANIMALS 10 multiparous sows. PROCEDURES Cytotoxic and CD4-CD8 double-positive T cells were separated from porcine colostrum and peripheral blood. Total RNA was extracted. The cDNA prepared from RNA was amplified, labeled, fragmented, and competitively hybridized to DNA microarray slides. The DNA microarray data were validated by use of a real-time reverse-transcription PCR assay, and expression of the genes FOS, NFKBI, IFNG, CXCR6, CCR5, ITGB2, CCR7, and SELL was assessed. Finally, DNA microarray data were validated at the protein level by use of flow cytometry via expression of c-Fos and integrin β-2. RESULTS Evaluation of gene expression profiles indicated that in contrast to results for peripheral blood, numerous cell-signaling pathways might be activated in colostrum. Profile analysis also revealed that FOS and NFKBI (genes of transcription factors) were involved in most cell-signaling pathways and that expression of these genes was significantly higher in colostral T cells than in peripheral blood T cells. Furthermore, CCR7 and SELL (genes of T-cell differentiation markers) in colostral T cells had expression patterns extremely similar to those found in effector or effector memory T cells. CONCLUSIONS AND CLINICAL RELEVANCE All or most of the T cells in colostrum had an effector-like phenotype and thus were more activated than those in peripheral blood. This gene expression profile would enable T cells to migrate to mammary glands, be secreted in colostrum, and likely contribute to passive immunity provided by sows to newborn pigs.

摘要

目的 比较猪初乳和外周血中T细胞的基因表达模式。动物 10头经产母猪。方法 从猪初乳和外周血中分离出细胞毒性T细胞和CD4 - CD8双阳性T细胞。提取总RNA。将从RNA制备的cDNA进行扩增、标记、片段化,并与DNA微阵列玻片进行竞争性杂交。通过实时逆转录PCR测定验证DNA微阵列数据,并评估FOS、NFKBI、IFNG、CXCR6、CCR5、ITGB2、CCR7和SELL基因的表达。最后,通过流式细胞术检测c - Fos和整合素β - 2的表达,在蛋白质水平验证DNA微阵列数据。结果 基因表达谱评估表明,与外周血结果相反,初乳中可能激活了众多细胞信号通路。谱分析还显示,FOS和NFKBI(转录因子基因)参与了大多数细胞信号通路,且这些基因在初乳T细胞中的表达显著高于外周血T细胞。此外,初乳T细胞中的CCR7和SELL(T细胞分化标志物基因)的表达模式与效应或效应记忆T细胞中的表达模式极为相似。结论与临床意义 初乳中所有或大多数T细胞具有类似效应细胞的表型,因此比外周血中的T细胞更具活性。这种基因表达谱使T细胞能够迁移至乳腺,分泌于初乳中,并可能有助于母猪为新生仔猪提供被动免疫。

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