Parturition in dairy cows temporarily alters the expression of genes in circulating neutrophils

Abstract

Extensive metabolic and physiologic changes occur during the peripartum, concurrent with a high incidence of infectious disease. Immune dysfunction is a likely contributor to the increased risk of disease at this time. Studies using high-yielding, total mixed ration-fed cows have indicated that neutrophil function is perturbed over the transition period; however, this reported dysfunction has yet to be investigated in moderate-yielding, grazing dairy cows. Therefore, we investigated changes in the expression of genes involved in neutrophil function. Blood was collected from cows at 5 time points over the transition period: precalving (-1wk; n=46), day of calving (d 0; n=46), and postcalving at wk 1 (n=46), wk 2 (n=45), and wk 4 (n=43). Neutrophils were isolated by differential centrifugation and gene expression was investigated. Quantitative reverse transcriptase PCR with custom-designed primer pairs and Roche Universal Probe Library (Roche, Basel, Switzerland) chemistry, combined with microfluidics integrated fluidic circuit chips (96.96 Dynamic Array, San Francisco, CA) were used to investigate the expression of 78 genes involved in neutrophil function and 18 endogenous control genes. Statistical significance between time points was determined using a repeated measures ANOVA. Genes that were differentially expressed over the transition period included those involved in neutrophil adhesion (SELL, ITGB2, and ITGBX), mediation of the immune response (TLR4, HLA-DRA, and CXCR2), maturation, cell cycle progression, apoptosis (MCL1, BCL2, FASLG, and RIPK1), and control of gene expression (PPARG, PPARD, and STAT3). We noted reduced gene expression of proinflammatory cytokines (IFNG, TNF, IL12, and CCL2) on the day of calving, whereas anti-inflammatory cytokine gene expression (IL10) was upregulated. Increased gene expression of antimicrobial peptides (BNBD4, DEFB10, and DEFB1) occurred on the day of calving. Collectively, transcription profiles are indicative of functional changes in neutrophils of grazing dairy cows over the transition period and align with studies in cows of conventional total mixed ration systems. This altered function may predispose cows to disease over the transition period and is likely to be a natural change in function due to parturition.

Effects of precalving body condition score and prepartum feeding level on production, reproduction, and health parameters in pasture-based transition dairy cows

“…Roche (2012) estimated that the cost of failing to transition optimally between pregnancy and lactation cost the New Zealand dairy industry in excess of $1.0 billion per annum. Immune competence is compromised during this period (Burton et al, 2001), even in low-to moderate-yielding grazing dairy cows (Crookenden et al, 2014;Heiser et al, 2015); this is likely a major reason for cows failing to transition and Effects of precalving body condition score and prepartum feeding level on production, reproduction, and health parameters in pasture-based transition dairy cows may be responsible for the estimated 30 to 50% of dairy cows that experience health disorders in the period immediately postcalving (LeBlanc, 2010).…”

Distinct neutrophil C5a receptor inflammatory events in cows initiated by chemoattractant C5a and lipopolysaccharide around parturition and in mid lactation

“…Caspase-3 is a key factor in apoptosis execution (Thornberry, 1998). Supporting evidence of this change in neutrophil apoptosis was also reported on the gene expression level by Madsen et al (2002) and Crookenden et al (2016). They reported a decreased expression in several proapoptotic genes with a concomitant increase in the expression of antiapoptotic genes on the day of parturition.…”

“…A possible explanation for the difference in viability between both lactation stages is that parturition primes the neutrophils, by which they get more sensitive to inflammatory stimuli. Stress, including parturition, increases the release of neutrophils from the bone marrow, followed by an increase in immature (band) cells in the circulation (Crookenden et al, 2016). Although our isolation protocol is set up to isolate mature neutrophils, a considerable percentage (25%) of the isolated neutrophils obtained from periparturient cows are band neutrophils.…”

“…Although our isolation protocol is set up to isolate mature neutrophils, a considerable percentage (25%) of the isolated neutrophils obtained from periparturient cows are band neutrophils. To a certain extent, these band neutrophils might also contribute to the increased viability observed in periparturient cells, which might be linked to the high cortisol levels at parturition (Burton et al, 1995;Crookenden et al, 2016). Although a shift in neutrophil population during the periparturient period could provide an explanation for the prolonged viability, the question remains whether the increased viability is solely due to this shift.…”

Effects of precalving body condition and prepartum feeding level on gene expression in circulating neutrophils

“…Immune function is altered during the transition period, which may contribute to the increased risk of health disorders (Detilleux et al, 1995;Mallard et al, 1998;Crookenden et al, 2016). It is increasingly evi-dent that prepartum nutrition and nutrient availability play an important role in the successful transition of dairy cows; for example, BCS and level of feeding affect indicators of health and metabolic function during the transition period (Roche et al, 2005(Roche et al, , 2013(Roche et al, , 2015Douglas et al, 2006;Loor et al, 2006).…”