Department of Range and Animal Sciences, New Mexico State University, Las Cruces, NM, 88003, USA.
Department of Animal and Food Sciences, Oklahoma State University, Stillwater, OK 74078, USA.
J Anim Sci. 2023 Jan 3;101. doi: 10.1093/jas/skad221.
Bovine respiratory disease (BRD) remains the greatest challenge facing the beef industry. Calves affected by BRD can manifest illness ranging from subclinical infection to acute death. In pathologies similar to BRD, extracellular histones have been implicated as major contributors to lung tissue damage. Histones are basic proteins responsible for DNA organization in cell nuclei, however when released extracellularly during cell injury or via neutrophil activation they become cytotoxic. Cattle suffering severe cases of BRD demonstrate reduced capacity to protect against the cytotoxic effects of histones, however, the protective mechanism(s) of serum remain(s) unknown. Therefore, the objective was to identify components within serum that contribute to protection against histone toxicity. Serum proteins from animals considered protective (P; N = 4) and nonprotective (NP; N = 4) against the toxic effects of histones were precipitated by the addition and incubation of exogenous histones. Proteins that interact with histones from both groups were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified via label free "shotgun" proteomics. Sixteen candidate proteins increased by ≥2-fold change in P vs. NP animals were identified, with several associated with the complement system. A subsequent study was conducted to evaluate complement system activity and serum protective capacity against exogenous histones in feedlot heifers. Serum samples were collected from 118 heifer calves (BW at arrival = 229 ± 2.4 kg) at feedlot arrival. Animals were retrospectively assigned to groups consisting of: calves not requiring treatment with antibiotics for BRD (CONT; N = 80), calves treated once (1TRT; N = 21), calves treated twice (2TRT; N = 5), calves treated thrice (3TRT; N = 3), or calves that died from BRD within 1 wk of entering the feedlot (DA; N = 9). Serum from DA animals was less protective than CONT (P = 0.0005) animals against histone toxicity. Complement activity of DA animals was reduced compared to CONT (P = 0.0044) animals. Additionally, the use of both assays as a ratio resulted in increased ability to detect DA animals. Results suggest that cattle predisposed to severe cases of respiratory disease may have impaired complement activity presumably contributing to reduced protective capacity against histone toxicity.
牛呼吸道疾病(BRD)仍然是牛肉行业面临的最大挑战。受 BRD 影响的牛犊可能会出现从亚临床感染到急性死亡等各种疾病。在与 BRD 相似的病理中,细胞外组蛋白已被认为是导致肺组织损伤的主要因素。组蛋白是负责细胞核内 DNA 组织的碱性蛋白,但在细胞损伤或中性粒细胞激活时释放到细胞外时,它们会变得具有细胞毒性。患有严重 BRD 的牛表现出降低的能力来抵抗组蛋白的细胞毒性作用,然而,血清的保护机制仍然未知。因此,本研究的目的是确定血清中有助于抵抗组蛋白毒性的成分。通过添加和孵育外源性组蛋白,将被认为对组蛋白毒性具有保护作用的(P;N=4)和非保护作用的(NP;N=4)动物的血清蛋白沉淀。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离与两组组蛋白相互作用的蛋白质,并通过无标记“shotgun”蛋白质组学进行鉴定。在 P 与 NP 动物中,有 16 种候选蛋白的表达增加了 2 倍以上,其中一些与补体系统有关。随后进行了一项研究,以评估育肥场小母牛的补体系统活性和血清对外源性组蛋白的保护能力。在育肥场到达时,从 118 头小母牛(到达时体重为 229±2.4kg)采集血清样本。动物回顾性地分为以下几组:不需要用抗生素治疗 BRD 的牛(CONT;N=80)、单次治疗的牛(1TRT;N=21)、两次治疗的牛(2TRT;N=5)、三次治疗的牛(3TRT;N=3)或在进入育肥场后 1 周内死于 BRD 的牛(DA;N=9)。与 CONT 动物(P=0.0005)相比,DA 动物的血清对组蛋白毒性的保护作用较差。与 CONT 动物(P=0.0044)相比,DA 动物的补体活性降低。此外,将两种检测方法作为比值使用可以提高检测 DA 动物的能力。结果表明,易患严重呼吸道疾病的牛可能补体活性受损,这可能导致其对组蛋白毒性的保护能力降低。