Treatment with granulocyte colony-stimulating factor has been reported to increase circulating neutrophil count and enhance neutrophil function in the periparturient cow. It was hypothesized that a commercially available recombinant bovine granulocyte colony-stimulating factor product (pegbovigrastim) affects gene expression profiles of neutrophils and supports neutrophil function in periparturient cows. Hence this study was undertaken to analyze expression of genes involved in neutrophil functions, including migration, interaction with pathogens, and cell survival. It also assessed the hypothesis that gene expression profiles in neutrophils are modulated by negative energy balance in the peripartum period. Holstein-Friesian, Jersey, and mixed-breed cows on pasture were blocked by expected calving date and body condition score and randomly assigned in a 2 × 2 factorial design. Cows were fed to exceed energy requirements prepartum (122%) or restricted to approximately 85% of prepartum energy requirements. At approximately 7 d before expected calving date, half the cows in each feed group were randomly assigned to be injected with pegbovigrastim or saline. Treatments were repeated within 24 h after calving. Blood samples were collected pretreatment approximately 7 d (d −7) before calving. Blood, uterine flush, and milk samples were collected at 4 (d 4) and 7 d in milk (d 7) to measure the expression of a panel of 20 genes representing cell adhesion, pattern recognition, inflammation and cytokine response, antimicrobial capacity, and apoptosis functions in neutrophils using NanoString technology (NanoString Technologies Inc., Seattle, WA) to quantify RNA copy numbers. No effects were observed of prepartum feeding group or a feeding group × treatment interaction for any of the investigated genes. An effect was observed of time on expression of several genes in blood neutrophils. After calving, expression of 2 of 4 cell adhesion-related genes, 3 of 4 pattern recognition receptors, 2 of 4 inflammatory genes, 2 antimicrobial genes, and 2 of 4 cell survival genes was significantly greater at d 4 or 7 or both compared with before calving (d −7). Expression of ICAM1, TLR2, and PTGS2 was significantly higher in blood neutrophils from animals treated with pegbovigrastim compared with untreated controls, suggesting greater migration, pattern recognition, and inflammatory response ability. Pegbovigrastim also affected RNA expression in uterine cells with ICAM1, NOD1, CLEC6A, PTGS2, MPO, DEFB5, and CATHL6 being expressed at higher levels and SELL, ITGB8, IL8RB, and IL10 at lower levels. Milk somatic cells showed a similar pattern but with fewer significant changes. In contrast to the reported decline in neutrophil function in the transition period, neutrophil gene expression was increased for many of the genes studied, an apparent attempt to compensate for reduced neutrophil function. Treatment with pegbovigrastim further increased expression of several genes involved in these processes in blood neutrophils and changed uterine cells to a phenotype with increased antimicrobial capacity, typical for neutrophils that have migrated into their target tissue.

据报道，用粒细胞集落刺激因子治疗可增加围产期母牛的循环中性粒细胞计数并增强中性粒细胞功能。假设市售重组牛粒细胞集落刺激因子产品（pegbovigrastim）影响中性粒细胞的基因表达谱并支持围产期母牛的中性​​粒细胞功能。因此，进行了这项研究以分析涉及嗜中性粒细胞功能的基因的表达，包括迁移，与病原体的相互作用和细胞存活。它还评估了以下假设：在围产期中性粒细胞中的基因表达谱受到负能量平衡的调节。荷斯坦-弗里斯兰，泽西岛，牧场上的奶牛和混种奶牛被预期的产犊日期和身体状况评分所阻止，并以2×2因子设计随机分配。母牛的摄食量超过了产前的能量需求（122％）或限制在产前能量需求的约85％。在预期产犊日期前约7天，将每个饲料组中的一半奶牛随机分配给匹格维格列汀或生理盐水注射。产犊后24小时内重复治疗。产犊前约7 d（d -7）预处理收集血样。在第4天（第4天）和第7天（第7天）收集血液，子宫冲洗液和牛奶样品，以测量一组20个基因的表达，这些基因代表细胞粘附，模式识别，炎症和细胞因子反应，抗菌能力，使用NanoString技术（NanoString Technologies Inc.，西雅图，华盛顿州）对嗜中性粒细胞的凋亡和凋亡功能进行定量。对于任何研究的基因，均未观察到产前喂养组或喂养组×治疗相互作用的影响。观察到时间对血液中性粒细胞中几个基因表达的影响。产犊后，第4天或第7天或两者相比，4个细胞粘附相关基因中的2个，4个模式识别受体中的3个，4个炎症基因中的2个，2个抗菌素基因和4个细胞存活基因中的2个的表达显着高于产犊前（d -7）。表达 观察到时间对血液中性粒细胞中几个基因表达的影响。产犊后，第4天或第7天或两者相比，4个细胞粘附相关基因中的2个，4个模式识别受体中的3个，4个炎症基因中的2个，2个抗菌素基因和4个细胞存活基因中的2个的表达显着高于产犊前（d -7）。表达 观察到时间对血液中性粒细胞中几个基因表达的影响。产犊后，第4天或第7天或两者相比，4个细胞粘附相关基因中的2个，4个模式识别受体中的3个，4个炎症基因中的2个，2个抗菌素基因和4个细胞存活基因中的2个的表达显着高于产犊前（d -7）。表达与未经治疗的对照组相比，接受培高斯汀治疗的动物的血液中性粒细胞中的ICAM1，TLR2和PTGS2显着更高，表明其迁移，模式识别和炎症反应能力更高。Pegbovigrastim也影响子宫细胞中RNA的表达，ICAM1，NOD1，CLEC6A，PTGS2，MPO，DEFB5和CATHL6的表达较高，而SELL，ITGB8，IL8RB和IL10在较低的水平。牛奶体细胞表现出相似的模式，但变化不大。与过渡期中性粒细胞功能下降的报道相反，许多研究的基因中性粒细胞基因表达增加，这显然是为了弥补中性粒细胞功能下降。pegbovigrastim的治疗进一步增加了涉及这些过程的几个基因在血液中性粒细胞中的表达，并使子宫细胞变为具有增加的抗菌能力的表型，这对于已迁移到其靶组织中的中性粒细胞是典型的。