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Generation of both MHC class I‐ and class II‐restricted antigenic peptides from exogenously added ovalbumin in murine phagosomes

机译:从小鼠吞噬体中外源性添加的卵清蛋白产生 MHC I 类和 II 类限制性抗原肽

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The phagosome fraction derived from a murine macrophage cell line (J774.1), which had internalized ovalbumin (OVA)-coated latex beads, was isolated. The peptides recovered from the phagosome fraction were separated on reverse phase HPLC and each fraction was analyzed for the content of either major histocompatibility complex (MHC) class I- or class II-restricted OVA-derived peptide. Both peptides were detected in the phagosome fraction after less than 15 min of internalization. It was also indicated that phagosomes degrade OVA protein into both MHC class I- and class II-restricted antigenic peptides by employing the same types of cathepsins. Furthermore, the results suggest that the MHC class I-restricted peptide rapidly exits from the phagosome to the cytosol. These findings illustrate a potential role for phagosomes not only in MHC class II-restricted but also in MHC class I-restricted exogenous antigen presentation pathways. Our results also point to the vital role of phagosomes in non-cytosolic antigen presentation pathway, in which further degradation of antigens by the proteasome is dispensable.
机译:分离出来自小鼠巨噬细胞系 (J774.1) 的吞噬体部分,该细胞系具有内化卵清蛋白 (OVA) 包被的乳胶珠。从吞噬体部分回收的肽在反相HPLC上分离,并分析每个组分的主要组织相容性复合物(MHC)I类或II类限制性OVA衍生肽的含量。在内化不到 15 分钟后在吞噬体部分中检测到两种肽。还表明,吞噬体通过使用相同类型的组织蛋白酶将 OVA 蛋白降解为 MHC I 类和 II 类限制性抗原肽。此外,结果表明,MHC I类限制性肽迅速从吞噬体退出到胞质溶胶。这些发现说明了吞噬体不仅在MHC II类限制性中,而且在MHC I类限制性外源性抗原呈递途径中的潜在作用。我们的研究结果还指出了吞噬体在非胞质抗原呈递途径中的重要作用,其中蛋白酶体进一步降解抗原是可有可无的。

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