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首页> 外文期刊>Science China Life Sciences >Cross-reactivity of anti-programmed death ligand 2 polyclonal antibody in mouse tissues
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Cross-reactivity of anti-programmed death ligand 2 polyclonal antibody in mouse tissues

机译：抗程序死亡配体2多克隆抗体在小鼠组织中的交叉反应

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The inhibitory co-receptor programmed death 1 (PD-1, encoded by pdcd1 ) and its two ligands PD-L1 and PD-L2 comprise an important immune inhibitory signaling pathway for defense against microbes and for self-tolerance. Unlike other members of the B7-CD28 family, expression of PD-L1 and PD-L2 is not limited to the immune system. In this study, we determined that a polyclonal antibody (pAb) (R&D Systems) against extracellular domains of mouse PD-L2 (mPD-L2) could recognize antigen(s) in diverse mouse tissues, including the anterior and intermediate pituitary gland, olfactory bulbs and olfactory epithelium, tongue epithelium, keratinized epithelial cells and skin and whisker hair follicles. These findings differed from previous reports of mPD-L2 localization. Reverse transcription PCR and Western blot analyses, however, were unable to detect any mPD-L2 transcripts or proteins of the 25-kD predicted molecular weight in RNA and protein extracts, respectively, from the above tissues, suggesting that the anti-mPD-L2 pAb cross-reacts with certain novel antigen(s). Developmental studies revealed that the earliest expression of mPD-L2-like antigen was in the olfactory epithelium at embryonic day 12.5 (E12.5). At E14.5, mPD-L2-like antigen was present in the skin, tongue and follicles of the skin and whiskers. The distribution patterns of mPD-L2-like antigen remained similar from E18.5 to adulthood. The results of bioinformatic analysis and other experiments suggested neural cell adhesion molecule and hemicentin-1 as candidate proteins with cross-reactivity to the anti-mPD-L2 pAb. These results demonstrate that care is required in interpreting staining patterns generated when anti-PD-L2 pAb is used to locate PD-L2-expressing cells in the central nervous system and epithelial tissues, such as the olfactory epithelium. In addition, this anti-PD-L2 pAb may be used as an alternative antibody for labeling the olfactory epithelium during embryonic development in mice.
机译：抑制性共受体程序性死亡1（PD-1，由pdcd1编码）及其两个配体PD-L1和PD-L2构成重要的免疫抑制信号传导途径，可防御微生物和实现自我耐受。与B7-CD28家族的其他成员不同，PD-L1和PD-L2的表达不限于免疫系统。在这项研究中，我们确定针对小鼠PD-L2（mPD-L2）胞外域的多克隆抗体（pAb）（R＆D Systems）可以识别多种小鼠组织中的抗原，包括垂体前叶和中间垂体，嗅觉鳞茎和嗅觉上皮，舌头上皮，角化上皮细胞以及皮肤和晶须的毛囊。这些发现与以前关于mPD-L2定位的报道有所不同。然而，逆转录PCR和蛋白质印迹分析无法从上述组织的RNA和蛋白质提取物中分别检测到任何mPD-L2转录本或25 kD预测分子量的蛋白质，这表明抗mPD-L2 pAb与某些新型抗原发生交叉反应。发育研究表明，在胚胎第12.5天（E12.5），mPD-L2样抗原的最早表达是在嗅觉上皮中。在E14.5，皮肤，舌头和毛囊以及晶须中存在mPD-L2样抗原。从E18.5到成年，mPD-L2样抗原的分布模式保持相似。生物信息学分析和其他实验的结果表明，神经细胞粘附分子和hemicentin-1是与抗mPD-L2 pAb具有交叉反应性的候选蛋白。这些结果表明，在解释使用抗PD-L2 pAb定位中枢神经系统和上皮组织（如嗅觉上皮）中的PD-L2表达细胞时产生的染色模式时需要格外小心。另外，该抗PD-L2 pAb可用作在小鼠胚胎发育过程中标记嗅觉上皮的替代抗体。

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