Changes in peripheral blood mononuclear cells’ mRNA expression of TLRs and CD14 during puerperal metritis in dairy cattle

Document Type: Full paper (Original article)

Authors

1 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran

2 Graduated from Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran

10.22099/ijvr.2020.33748.5008

Abstract

Background: Peripheral blood mononuclear cells (PBMCs), commonly referred to as lymphocytes and monocytes, representing cells of the innate and adaptive immune systems. Aims: To find out whether changes in PBMCs’ mRNA expression of pattern recognition receptors (PRRs) are associated with puerperal metritis in Holstein cows. Methods:‏ Peripheral blood mononuclear cells were collected from 20 cows with puerperal metritis and 20 cows without metritis at 10 days postpartum. Expression of toll-like receptors 2 and 4 (TLR2 and TLR4), and cluster of differentiation 14 (CD14) genes were assessed in PBMCs using a quantitative real time-polymerase chain reaction (qRT-PCR) technique. The data was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a reference gene, and 2-∆∆Ct methodology was used for relative quantification. Results: The results of the present study demonstrated that the expression of TLR4 (P=0.04) and CD14 (P=0.008) was significantly greater in cows with puerperal metritis compared to the control group. However, the expression of TLR2 (P=0.06) was not significantly different between cows with puerperal metritis and healthy cows. Conclusion: This study suggests that puerperal metritis significantly increases the expression of TLR4 and CD14 genes in the PBMCs which contributes to the proper stimulation of inflammation and uterine clearance of bacteria soon after calving.

Keywords

Main Subjects


Ahmadi, M; Makki, M; Mirzaei, A and Gheisari, H (2019). Effects of hypertonic dextrose and paraffin solution as non- antibiotic treatments of clinical endometritis on reproductive performance of high producing dairy cows. Reprod. Domest. Anim., 54: 762-771.

Bilal, MS; Abaker, JA; Aabdin, ZU; Xu, T; Dai, H; Zhang, K; Liu, X and Shen, X (2016). Lipopolysaccharide derived from the digestive tract triggers an inflammatory response in the uterus of mid-lactating dairy cows during SARA. BMC Vet. Res., 12: 284-293.

Brubaker, SW; Bonham, KS; Zanoni, I and Kagan, JC (2015). Innate immune pattern recognition: a cell biological perspective. Annu. Rev. Immunol., 33: 257-290.

Cai, TQ; Weston, P; Lund, L; Brodie, B; McKenna, D and Wagner, W (1994). Association between neutrophil functions and periparturient disorders in cows. Am. J. Vet. Res., 55: 934-943.

Cunha, F; Jeon, SJ; Daetz, R; Vieira-Neto, A; Laporta, J; Jeong, KC; Barbet, AF; Risco, CA and Galvao, KN (2018). Quantifying known and emerging uterine pathogens, and evaluating their association with metritis and fever in dairy cows. Theriogenology. 114: 25-33.

da Mata Martins, T; da Paixão, TA; Costa, ÉA; de Carvalho Pires, A; Santos, RL and Borges, ÁM (2011). Postpartum toll-like receptors and β-defensin 5 mRNA levels in the endometrium of Holstein cows. Vet. Immunol. Immunopathol., 139: 277-281.

Davies, D; Meade, KG; Herath, S; Eckersall, PD; Gonzalez, D; White, JO; Conlan, RS; O’Farrelly, C and Sheldon, IM (2008). Toll-like receptor and antimicrobial peptide expression in the bovine endometrium. Reprod. Biol. Endocrinol., 6: 53.

Eckel, EF and Ametaj, BN (2016). Role of bacterial endotoxins in the etiopathogenesis of periparturient diseases of transition dairy cows. J. Dairy Sci., 99: 5967-5990.

Fischer, C; Drillich, M; Odau, S; Heuwieser, W; Einspanier, R and Gabler, C (2010). Selected pro-inflammatory factor transcripts in bovine endometrial epithelial cells are regulated during the oestrous cycle and elevated in case of subclinical or clinical endometritis. Reprod. Fertil. Dev., 22: 818-829.

Fu, Y; Liu, B; Feng, X; Liu, Z; Liang, D; Li, F; Li, D; Cao, Y; Feng, S and Zhang, X (2013). Lipopolysaccharide increases toll-like receptor 4 and downstream toll-like receptor signaling molecules expression in bovine endometrial epithelial cells. Vet. Immunol. Immunopathol., 151: 20-27.

Herath, S; Dobson, H; Bryant, C and Sheldon, I (2006a). Use of the cow as a large animal model of uterine infection and immunity. J. Reprod. Immunol., 69: 13-22.

Herath, S; Fischer, DP; Werling, D; Williams, EJ; Lilly, ST; Dobson, H; Bryant, CE and Sheldon, IM (2006b). Expression and function of toll-like receptor 4 in the endometrial cells of the uterus. Endocrinology. 147: 562-570.

Herath, S; Lilly, ST; Santos, NR; Gilbert, RO; Goetze, L; Bryant, CE; White, JO; Cronin, J and Sheldon, IM (2009). Expression of genes associated with immunity in the endometrium of cattle with disparate postpartum uterine disease and fertility. Reprod. Biol. Endocrinol., 7: 55-88.

Kawai, T and Akira, S (2010). The rule of pattern recognition receptors in innate immunity: update on toll-like receptors. Nat. Immunol., 11: 327-384.

Kim, IH; Na, KJ and Yang, MP (2005). Immune responses during the peripartum period in dairy cows with postpartum endometritis. J. Reprod. Develop. 6: 757-764.

Koneman, EW; Minckler, TM; Shires, DB and De Jonch, DS (1971). Postmortem bacteriology: II. Selection of cases for culture. Am. J. Clin. Pathol., 55: 17-23.

Lange, J; McCarthy, A; Kay, J; Meier, S; Walker, C; Crookenden, MA; Mitchell, MD, Loor, JJ; Roche, JR and Heiser, A (2016). Prepartum feeding level and body condition score affect immunological performance in grazing dairy cows during the transition period. J. Dairy Sci., 99: 2329-2338.

Mahnani, A; Sadeghi-Sefidmazgi, A and Cabrera, VE (2015). Consequences and economics of metritis in Iranian Holstein dairy farms. J. Dairy Sci., 98: 6048-6057.

Miyake, K (2006). Invited review: roles for accessory molecules in microbial recognition by toll-like receptors. J. Endotoxin Res., 12: 195-204.

O’Neill, LA; Bryant, CE and Doyle, SL (2009). Therapeutic targeting of toll-like receptors for infectious and inflammatory diseases and cancer. Pharmacol. Rev., 61: 1073-1094.

Reppert, EJ (2015). Evidence for the use of ceftiofur for treatment of metritis in dairy cattle. Vet. Clin. Anim. Pract., 31: 139-149.

Schöniger, S; Gräfe, H and Schoon, HA (2017). Expression of toll-like receptors 2, 4 and 6 in different cell populations of the equine endometrium. Vet. Immunol. Immunopathol., 185: 7-13.

Sheldon, IM and Dobson, H (2004). Postpartum uterine health in cattle. Anim. Reprod. Sci., 82: 295-306.

Sheldon, IM; Lewis, GS; LeBlanc, S and Gilbert, RO (2006). Defining postpartum uterine disease in cattle. Theriogenology. 65: 1516-1530.

Sheldon, IM; Noakes, D; Rycroft, A; Pfeiffer, D and Dobson, H (2002). Influence of uterine bacterial contamination after parturition on ovarian dominant follicle selection and follicle growth and function in cattle. Reproduction. 123: 837-845.

Sheldon, IM; Rycroft, AN; Dogan, B; Craven, M; Bromfield, JJ; Chandler, A; Roberts, MH; Price, SB; Gilbert, RO and Simpson, KW (2010). Specific strains of Escherichia coli are pathogenic for the endometrium of cattle and cause pelvic inflammatory disease in cattle and mice. Plos One. 5: e9192.

Swangchan-Uthai, T; Lavender, CR; Cheng, Z; Fouladi-Nashta, AA and Wathes, DC (2012). Time course of defense mechanisms in bovine endometrium in response to lipopolysaccharide. Biol. Reprod., 87: 131-113, 135.

Takeuchi, O and Akira, S (2010). Pattern recognition receptors and inflammation. Cell. 140: 805-820.

Takeuchi, O; Hoshino, K and Akira, S (2000). Cutting edge: TLR2-deficient and MyD88-deficient mice are highly susceptible to Staphylococcus aureus infection. J. Immunol., 165: 5392-5396.

Triantafilou, M and Triantafilou, K (2002). Lipopoly-saccharide recognition: CD14, TLRs and the LPS-activation cluster. Trends Immunol., 23: 301-304.

van der Molen, RG; Schutten, JHF; van Cranenbroek, B; ter Meer, M; Donckers, J; Scholten, RR; van der Heijden, OWH; Spaanderman, MEA and Joosten, I (2014). Menstrual blood closely resembles the uterine immune micro-environment and is clearly distinct from peripheral blood. Hum. Reprod., 29: 303-314.

Williams, EJ; Fischer, DP; Pfeiffer, DU; England, GC; Noakes, DE; Dobson, H and Sheldon, IM (2005). Clinical evaluation of postpartum vaginal mucus reflects uterine bacterial infection and the immune response in cattle. Theriogenology. 63: 102-117.

Yapan, S; Liping, S; Aizhen, G and Liguo, Y (2014). Effects of toll-like receptor 2 gene mutation on resistance to bovine brucellosis. Livest. Sci., 170: 30-34.

Yoshimura, A; Lien, E; Ingalls, R; Heine, H; Henneke, P; Teti, G; Espevik, T; Kato, I and Golenbock, D (2000). Recognition of gram-positive bacterial cell wall components by the innate immune system occurs via toll-like receptor 2. J. Endotoxin Res., 6: S13-S13.

Zähringer, U; Lindner, B; Inamura, S; Heine, H and Alexander, C (2008). TLR2-promiscuous or specific? A critical re-evaluation of a receptor expressing apparent broad specificity. Immunobiology. 213: 205-224.