title: 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.
authors.affiliation: DairyNZ, c/o University of Auckland, Auckland 1010, New Zealand; Institute of Vet, Animal and Biomedical Sciences, Massey University, Palmerston North 4442, New Zealand. Electronic address: mallory.crookenden@dairynz.co.nz.
authors.forename: M A
authors.initials: MA
authors.lastname: Crookenden
authors.affiliation: Institute of Vet, Animal and Biomedical Sciences, Massey University, Palmerston North 4442, New Zealand; AgResearch, Hopkirk Research Institute, Palmerston North 4442, New Zealand.
authors.forename: A
authors.initials: A
authors.lastname: Heiser
authors.affiliation: Institute of Vet, Animal and Biomedical Sciences, Massey University, Palmerston North 4442, New Zealand.
authors.forename: A
authors.initials: A
authors.lastname: Murray
authors.affiliation: Institute of Vet, Animal and Biomedical Sciences, Massey University, Palmerston North 4442, New Zealand.
authors.forename: V S R
authors.initials: VSR
authors.lastname: Dukkipati
authors.affiliation: DairyNZ, Newstead, Hamilton 3284, New Zealand.
authors.forename: J K
authors.initials: JK
authors.lastname: Kay
authors.affiliation: Department of Animal Sciences, University of Illinois, Urbana 61801.
authors.forename: J J
authors.initials: JJ
authors.lastname: Loor
authors.affiliation: DairyNZ, Newstead, Hamilton 3284, New Zealand.
authors.forename: S
authors.initials: S
authors.lastname: Meier
authors.affiliation: University of Queensland, Centre for Clinical Research, Herston, Queensland 4029, Australia.
authors.forename: M D
authors.initials: MD
authors.lastname: Mitchell
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland, College Park 20742.
authors.forename: K M
authors.initials: KM
authors.lastname: Moyes
authors.affiliation: DairyNZ, c/o University of Auckland, Auckland 1010, New Zealand.
authors.forename: C G
authors.initials: CG
authors.lastname: Walker
authors.affiliation: DairyNZ, Newstead, Hamilton 3284, New Zealand.
authors.forename: J R
authors.initials: JR
authors.lastname: Roche
mesh_terms: D000818:Animals; D002417:Cattle; D016207:Cytokines / Q000378:metabolism; D004032:Diet / Q000662:veterinary; D005260:Female; D015870:Gene Expression; D007774:Lactation; D009504:Neutrophils / Q000276:immunology*; D036801:Parturition; D058725:Peripartum Period
chemical_list: D016207:Cytokines
keywords: Fluidigm; PMN; innate immune function; peripartum period