Ortín-Bustillo Alba, Martínez-Subiela Silvia, Cerón José Joaquín, González-Bulnes Antonio, Tvarijonaviciute Asta, Muñoz-Prieto Alberto
Salilab-UMU, Interdisciplinary Laboratory of Clinical Analysis (Interlab), Regional Campus of International Excellence 'Campus Mare Nostrum', University of Murcia, Murcia, Spain.
Department of Animal Health and Production, Faculty of Veterinary, Universidad Cardenal Herrera-CEU, CEU Universities, C/Tirant lo Blanc, 7, Alfara del Patriarca, Valencia, Spain.
PLoS One. 2025 Oct 24;20(10):e0335133. doi: 10.1371/journal.pone.0335133. eCollection 2025.
Inter-alpha-trypsin inhibitor protein heavy chain 4 (ITIH4), also named porcine major acute phase protein (Pig-MAP), is a positive acute phase protein (APP) in pigs and can be measured in plasma and also in saliva to assess the animal health. The objectives of this report were (1) to evaluate if different commercially available porcine ELISA kits can measure ITIH4 concentrations in saliva samples of pigs, and (2) to study the distribution of ITIH4 components in saliva and compare it to the distribution in plasma. The results showed that two of the ELISA kits used in this report could measure salivary ITIH4 with precision and accuracy, but only one showed significant differences between pigs with tail biting and control animals without this condition. Western blotting analysis revealed the presence of a different distribution of ITIH4 bands in saliva samples compared to plasma samples. In conclusion, in saliva of pigs ITIH4 can be quantified using a commercial ELISA kit increasing its concentration in cases of tail biting. In addition, ITIH4 shows bands at western blot in saliva that are different from serum but are compatible with different ITIH4 forms. These data confirm that ITIH4 can be detected in saliva and be potentially used as a biomarker of inflammation.
α-胰蛋白酶抑制剂重链4（ITIH4），也称为猪主要急性期蛋白（Pig-MAP），是猪体内的一种阳性急性期蛋白（APP），可在血浆和唾液中检测，用于评估动物健康状况。本报告的目的是：（1）评估不同市售猪ELISA试剂盒能否检测猪唾液样本中的ITIH4浓度；（2）研究ITIH4成分在唾液中的分布，并与血浆中的分布进行比较。结果表明，本报告中使用的两种ELISA试剂盒能够精确、准确地检测唾液中的ITIH4，但只有一种试剂盒显示，咬尾猪与无此情况的对照动物之间存在显著差异。蛋白质印迹分析显示，与血浆样本相比，唾液样本中ITIH4条带的分布有所不同。总之，在猪的唾液中，ITIH4可以使用商业ELISA试剂盒进行定量检测，在咬尾情况下其浓度会升高。此外，ITIH4在唾液蛋白质印迹中显示出与血清不同的条带，但与不同的ITIH4形式相符。这些数据证实，ITIH4可以在唾液中检测到，并有可能用作炎症的生物标志物。
