■妊娠结束,随后分娩,新生儿期代表了母亲和后代免疫学变化的高度动态阶段。妊娠晚期母胎界面先天免疫细胞的调节,出生后,在新生儿肠道和其他粘膜表面的微生物定植过程中,对于控制炎症和维持体内平衡至关重要。固有免疫细胞和粘膜上皮细胞表达抗白细胞蛋白酶(SLPI),具有抗炎和抗蛋白酶活性,可以调节细胞活化。
■这里,我们首次在马中开发并验证了新的单克隆抗体(mAb)来表征SLPI.分娩后直接从健康的成年马和一群母马及其小马驹中收集外周血和粘膜样本,以评估这一关键阶段。
■首先,我们通过流式细胞术定义了外周血中产生SLPI的细胞类型,突出显示中性粒细胞和CD14+单核细胞的子集作为SLPI分泌免疫细胞。使用新的SLPImAb开发了基于荧光珠的测定,并用于建立血清和粘膜表面分泌样品中分泌的SLPI的基线浓度。包括唾液,鼻腔分泌物,初乳,和牛奶。这证明了SLPI在各种马组织中的组成型分泌,包括高初乳浓度。使用免疫荧光,我们确定了粘膜组织中SLPI的产生。最后,对临床健康的母马和小马驹进行纵向采样,可以监测血清SLPI浓度.在新生儿和产后母马中,SLPI在分娩当天达到顶峰,母马在一周内恢复成年正常,小马驹在三个月大之前保持明显较高的SLPI分泌。
■这证明了母马及其小马驹的SLPI的生理系统变化,特别是在出生时,在这个关键时期,可能有助于调节先天免疫反应。
UNASSIGNED: The end of gestation, ensuing parturition, and the neonatal period represent highly dynamic phases for immunological changes in both mother and offspring. The regulation of innate immune cells at the maternal-fetal interface during late term pregnancy, after birth, and during microbial colonization of the neonatal gut and other mucosal surfaces, is crucial for controlling inflammation and maintaining homeostasis. Innate immune cells and mucosal epithelial cells express antileukoproteinase (SLPI), which has anti-inflammatory and anti-protease activity that can regulate cellular activation.
UNASSIGNED: Here, we developed and validated new monoclonal antibodies (mAbs) to characterize SLPI for the first time in horses. Peripheral blood and mucosal samples were collected from healthy adults horses and a cohort of mares and their foals directly following parturition to assess this crucial stage.
UNASSIGNED: First, we defined the cell types producing SLPI in peripheral blood by flow cytometry, highlighting the neutrophils and a subset of the CD14+ monocytes as SLPI secreting immune cells. A fluorescent bead-based assay was developed with the new SLPI mAbs and used to establish baseline concentrations for secreted SLPI in serum and secretion samples from mucosal surfaces, including saliva, nasal secretion, colostrum, and milk. This demonstrated constitutive secretion of SLPI in a variety of equine tissues, including high colostrum concentrations. Using immunofluorescence, we identified production of SLPI in mucosal tissue. Finally, longitudinal sampling of clinically healthy mares and foals allowed monitoring of serum SLPI concentrations. In neonates and postpartum mares, SLPI peaked on the day of parturition, with mares returning to the adult normal within a week and foals maintaining significantly higher SLPI secretion until three months of age.
UNASSIGNED: This demonstrated a physiological systemic change in SLPI in both mares and their foals, particularly at the time around birth, likely contributing to the regulation of innate immune responses during this critical period.