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能够识别质谱法无法检测到的肽污染物的 T 细胞。

T cells recognizing a peptide contaminant undetectable by mass spectrometry.

机构信息

INSERM, U986, DeAR Lab Avenir, Saint Vincent de Paul Hospital, Paris, France.

出版信息

PLoS One. 2011;6(12):e28866. doi: 10.1371/journal.pone.0028866. Epub 2011 Dec 14.

DOI:10.1371/journal.pone.0028866
PMID:22194932
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3237501/
Abstract

Synthetic peptides are widely used in immunological research as epitopes to stimulate their cognate T cells. These preparations are never completely pure, but trace contaminants are commonly revealed by mass spectrometry quality controls. In an effort to characterize novel major histocompatibility complex (MHC) Class I-restricted β-cell epitopes in non-obese diabetic (NOD) mice, we identified islet-infiltrating CD8+ T cells recognizing a contaminating peptide. The amount of this contaminant was so small to be undetectable by direct mass spectrometry. Only after concentration by liquid chromatography, we observed a mass peak corresponding to an immunodominant islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP)(206-214) epitope described in the literature. Generation of CD8+ T-cell clones recognizing IGRP(206-214) using a novel method confirmed the identity of the contaminant, further underlining the immunodominance of IGRP(206-214). If left undetected, minute impurities in synthetic peptide preparations may thus give spurious results.

摘要

合成肽被广泛用于免疫研究，作为刺激其同源 T 细胞的表位。这些制剂从来都不是完全纯净的，但痕量污染物通常会通过质谱质量控制来揭示。为了鉴定非肥胖型糖尿病 (NOD) 小鼠中新的主要组织相容性复合物 (MHC) Ⅰ类限制性β细胞表位，我们鉴定了识别浸润胰岛的 CD8+T 细胞的一个污染肽。这种污染物的量太小，无法通过直接质谱检测到。只有经过液相色谱浓缩后，我们才观察到与文献中描述的免疫优势胰岛特异性葡萄糖-6-磷酸酶催化亚基相关蛋白 (IGRP)(206-214) 表位相对应的质量峰。使用一种新方法生成识别 IGRP(206-214)的 CD8+T 细胞克隆，证实了污染物的身份，进一步强调了 IGRP(206-214)的免疫优势。如果不加以检测，合成肽制剂中的微量杂质可能会导致虚假结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/dad347d40f94/pone.0028866.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/484577626a08/pone.0028866.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/b85ad40ceed3/pone.0028866.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/adecaf721dbb/pone.0028866.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/32d64dc124df/pone.0028866.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/31de8e1cc218/pone.0028866.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/dad347d40f94/pone.0028866.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/484577626a08/pone.0028866.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/b85ad40ceed3/pone.0028866.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/adecaf721dbb/pone.0028866.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/32d64dc124df/pone.0028866.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/31de8e1cc218/pone.0028866.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc6d/3237501/dad347d40f94/pone.0028866.g006.jpg

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Antigen-based immune therapeutics for type 1 diabetes: magic bullets or ordinary blanks?用于1型糖尿病的基于抗原的免疫疗法：神奇子弹还是普通空白？
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