Oestrous cycle-dependent equine uterine immune response to induced infectious endometritis.
Abstract: Infectious endometritis is a major cause of reduced pregnancy rates in horses. The objectives of this study were to establish a timeline of the innate immune response in the uterus of healthy horses and to investigate the oestrous cycle effect on this. Endometrial biopsies were collected from five horses before and at 3, 12, 24, 48 and 72 h after inoculation of Escherichia coli, once in oestrus and once in dioestrus. They were analysed by quantitative real-time PCR, microbiology and histology. Neutrophil numbers increased from very low levels in the absence of inflammation to severe neutrophilia 3 h after inoculation. The concentrations of mRNAs for Toll-like receptor (TLR)2, TLR4, NOD-like receptor NLRC5, tissue inhibitor of metallopeptidases 1 (TIMP1) and chemokines CCL2, CXCL9, CXCL10 and CXCL11 were all increased 3 h after inoculation of E. coli compared to levels detected prior to inoculation. Chemokine mRNA levels remained elevated for 48 h. Concentrations of mRNAs for the antimicrobial peptides equine β-defensin 1 (EBD1), lysozyme, secretory leukoprotease inhibitor (SLPI), lipocalin 2 (LCN2), lactoferrin and uteroferrin were increased between 3 and 12 h post inoculation. The gene for secreted phospholipase A (sPLA) was expressed constitutively. P19 uterocalin mRNA levels were higher in dioestrus than in oestrus over the first 24 h of inflammation. Neutrophils and many innate immune genes responded rapidly to the introduction of E. coli into the uterus, while the oestrous cycle stage had only a relatively minor effect on the response to E. coli. This study has delineated a useful model of innate immunity in infectious endometritis of healthy animals.
Publication Date: 2016-11-08 PubMed ID: 27825391PubMed Central: PMC5101692DOI: 10.1186/s13567-016-0398-xGoogle Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research article discusses a study carried out to examine the timeline of the uterus’s immune response in healthy horses, focusing on the influence of the oestrous cycle. The aim was to understand better the body’s defense mechanism against infectious endometritis, a significant cause of reduced pregnancy rates in horses.
Objective and Methodology
- The primary aim of this research was to trace the timeframe of the immune response in the uterus of healthy horses following an induced endometritis, an inflammation of the inner lining of the uterus.
- Another objective was to explore the effect of the oestrous cycle, the reproductive cycle in horses, on this immune response.
- To achieve this, endometrial biopsies were taken from five horses at different times after the inoculation of Escherichia coli, a common bacterium known to cause endometritis. The biopsies were obtained once in oestrus and once in dioestrus, the two main stages of the horses’ oestrous cycle.
- The biopsies were then analysed using quantitative real-time PCR, a technology used for detecting and quantifying specific DNA sequences, along with standard microbiological testing and histological analysis, which involves the examination of biological tissues to observe the appearance of diseased cells and tissues in very fine detail.
Findings
- The researchers documented a significant increase in neutrophil numbers, a white blood cell that helps fight infection, from very low levels without inflammation to severe neutrophilia (an increase in neutrophils) within 3 hours after inoculation with E. coli.
- They reported an increased expression of various innate immune genes and chemokines, which are proteins that induce directed chemotaxis (movement of an organism in response to a chemical stimulus) in nearby responsive cells, 3 hours after the inoculation compared to pre-inoculation levels.
- The levels of the antimicrobial peptides, which are short proteins capable of killing or inhibiting the growth of microorganisms, also increased between 3 and 12 hours post-inoculation.
- The research indicated an relatively minor effect of the oestrous cycle stage on the immune response to E. coli.
Conclusion
- The study provided a practical model of innate immunity in infectious endometritis among healthy horses. Understanding this timeline of immune response, as well as the differing responses at different stages of the oestrous cycle, can significantly contribute to improved management and treatment of endometritis in horses.
Cite This Article
APA
Marth CD, Firestone SM, Glenton LY, Browning GF, Young ND, Krekeler N.
(2016).
Oestrous cycle-dependent equine uterine immune response to induced infectious endometritis.
Vet Res, 47(1), 110.
https://doi.org/10.1186/s13567-016-0398-x Publication
Researcher Affiliations
- Asia-Pacific Centre for Animal Health, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, VIC, Australia. christina.marth@unimelb.edu.au.
- Translational Research and Animal Clinical Trial Study Group, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Werribee, VIC, Australia. christina.marth@unimelb.edu.au.
- Asia-Pacific Centre for Animal Health, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, VIC, Australia.
- Asia-Pacific Centre for Animal Health, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, VIC, Australia.
- Translational Research and Animal Clinical Trial Study Group, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Werribee, VIC, Australia.
- Asia-Pacific Centre for Animal Health, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, VIC, Australia.
- Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, VIC, Australia.
- Asia-Pacific Centre for Animal Health, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, VIC, Australia.
- Translational Research and Animal Clinical Trial Study Group, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Werribee, VIC, Australia.
MeSH Terms
- Animals
- Chemokines / physiology
- Endometritis / immunology
- Endometritis / microbiology
- Endometritis / veterinary
- Escherichia coli Infections / immunology
- Escherichia coli Infections / microbiology
- Escherichia coli Infections / veterinary
- Estrous Cycle / physiology
- Female
- Horse Diseases / immunology
- Horse Diseases / microbiology
- Horses
- Immunity, Innate / immunology
- Immunity, Innate / physiology
- Real-Time Polymerase Chain Reaction / veterinary
- Toll-Like Receptors / physiology
- Uterus / immunology
- Uterus / physiopathology
References
This article includes 63 references
- Zent WW, Troedsson MHT, Xue J-L. Postbreeding uterine fluid accumulation in a normal population of Thoroughbred mares: a field study. Proceedings of the annual convention of the American Association of Equine Practitioners 1998.
- Riddle WT, LeBlanc MM, Stromberg AJ. Relationships between uterine culture, cytology and pregnancy rates in a Thoroughbred practice.. Theriogenology 2007 Aug;68(3):395-402.
- Troedsson MH, Loset K, Alghamdi AM, Dahms B, Crabo BG. Interaction between equine semen and the endometrium: the inflammatory response to semen.. Anim Reprod Sci 2001 Dec 3;68(3-4):273-8.
- Watson ED. Post-breeding endometritis in the mare.. Anim Reprod Sci 2000 Jul 2;60-61:221-32.
- Fumuso EA, Aguilar J, Giguère S, Rivulgo M, Wade J, Rogan D. Immune parameters in mares resistant and susceptible to persistent post-breeding endometritis: effects of immunomodulation.. Vet Immunol Immunopathol 2007 Jul 15;118(1-2):30-9.
- Woodward EM, Christoffersen M, Campos J, Betancourt A, Horohov D, Scoggin KE, Squires EL, Troedsson MH. Endometrial inflammatory markers of the early immune response in mares susceptible or resistant to persistent breeding-induced endometritis.. Reproduction 2013 Mar 1;145(3):289-96.
- Christoffersen M, Woodward E, Bojesen AM, Jacobsen S, Petersen MR, Troedsson MH, Lehn-Jensen H. Inflammatory responses to induced infectious endometritis in mares resistant or susceptible to persistent endometritis.. BMC Vet Res 2012 Mar 29;8:41.
- Bain AM. The rôle of infection in infertility in the thoroughbred mare.. Vet Rec 1966 Jan 29;78(5):168-73.
- Collins S. A study of the incidence of cervical and uterine infection in Thoroughbred mares in Ireland. Vet Rec 1964;66:673–676.
- Albihn A, Båverud V, Magnusson U. Uterine microbiology and antimicrobial susceptibility in isolated bacteria from mares with fertility problems.. Acta Vet Scand 2003;44(3-4):121-9.
- Nikolakopoulos E, Watson ED. Uterine contractility is necessary for the clearance of intrauterine fluid but not bacteria after bacterial infusion in the mare.. Theriogenology 1999 Aug;52(3):413-23.
- Troedsson MH, Liu IK, Thurmond M. Function of uterine and blood-derived polymorphonuclear neutrophils in mares susceptible and resistant to chronic uterine infection: phagocytosis and chemotaxis.. Biol Reprod 1993 Sep;49(3):507-14.
- Troedsson MH, Liu IK, Ing M, Pascoe J, Thurmond M. Multiple site electromyography recordings of uterine activity following an intrauterine bacterial challenge in mares susceptible and resistant to chronic uterine infection.. J Reprod Fertil 1993 Nov;99(2):307-13.
- LeBlanc MM, Neuwirth L, Asbury AC, Tran T, Mauragis D, Klapstein E. Scintigraphic measurement of uterine clearance in normal mares and mares with recurrent endometritis.. Equine Vet J 1994 Mar;26(2):109-13.
- Evans MJ, Hamer JM, Gason LM, Graham CS, Asbury AC, Irvine CH. Clearance of bacteria and non-antigenic markers following intra-uterine inoculation into maiden mares: Effect of steroid hormone environment.. Theriogenology 1986 Jul;26(1):37-50.
- Marth CD, Young ND, Glenton LY, Noden DM, Browning GF, Krekeler N. Effect of ovarian hormones on the healthy equine uterus: a global gene expression analysis.. Reprod Fertil Dev 2015 May 20;.
- Christoffersen M, Baagoe CD, Jacobsen S, Bojesen AM, Petersen MR, Lehn-Jensen H. Evaluation of the systemic acute phase response and endometrial gene expression of serum amyloid A and pro- and anti-inflammatory cytokines in mares with experimentally induced endometritis.. Vet Immunol Immunopathol 2010 Nov 15;138(1-2):95-105.
- Marth CD, Young ND, Glenton LY, Noden DM, Browning GF, Krekeler N. Deep sequencing of the uterine immune response to bacteria during the equine oestrous cycle.. BMC Genomics 2015 Nov 14;16:934.
- Atli M, Kurar E, Kayis S, Aslan S, Semacan A, Celik S, Guzeloglu A. Expression of Toll-like receptors (TLRs) in the equine endometrium during the estrous cycle. Reprod Domest Anim 2010;45:58.
- Chow JC, Young DW, Golenbock DT, Christ WJ, Gusovsky F. Toll-like receptor-4 mediates lipopolysaccharide-induced signal transduction.. J Biol Chem 1999 Apr 16;274(16):10689-92.
- Hoshino K, Takeuchi O, Kawai T, Sanjo H, Ogawa T, Takeda Y, Takeda K, Akira S. Cutting edge: Toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysaccharide: evidence for TLR4 as the Lps gene product.. J Immunol 1999 Apr 1;162(7):3749-52.
- Matsuguchi T, Musikacharoen T, Ogawa T, Yoshikai Y. Gene expressions of Toll-like receptor 2, but not Toll-like receptor 4, is induced by LPS and inflammatory cytokines in mouse macrophages.. J Immunol 2000 Nov 15;165(10):5767-72.
- Barbé F, Douglas T, Saleh M. Advances in Nod-like receptors (NLR) biology.. Cytokine Growth Factor Rev 2014 Dec;25(6):681-97.
- Zlotnik A, Yoshie O. The chemokine superfamily revisited.. Immunity 2012 May 25;36(5):705-16.
- Cole AM, Ganz T, Liese AM, Burdick MD, Liu L, Strieter RM. Cutting edge: IFN-inducible ELR- CXC chemokines display defensin-like antimicrobial activity.. J Immunol 2001 Jul 15;167(2):623-7.
- Yang D, Chen Q, Hoover DM, Staley P, Tucker KD, Lubkowski J, Oppenheim JJ. Many chemokines including CCL20/MIP-3alpha display antimicrobial activity.. J Leukoc Biol 2003 Sep;74(3):448-55.
- Bruhn O, Grötzinger J, Cascorbi I, Jung S. Antimicrobial peptides and proteins of the horse--insights into a well-armed organism.. Vet Res 2011 Sep 2;42(1):98.
- Linde A, Ross CR, Davis EG, Dib L, Blecha F, Melgarejo T. Innate immunity and host defense peptides in veterinary medicine.. J Vet Intern Med 2008 Mar-Apr;22(2):247-65.
- Nevalainen TJ, Graham GG, Scott KF. Antibacterial actions of secreted phospholipases A2. Review.. Biochim Biophys Acta 2008 Jan-Feb;1781(1-2):1-9.
- Tomee JF, Koëter GH, Hiemstra PS, Kauffman HF. Secretory leukoprotease inhibitor: a native antimicrobial protein presenting a new therapeutic option?. Thorax 1998 Feb;53(2):114-6.
- Couto MA, Harwig SS, Cullor JS, Hughes JP, Lehrer RI. eNAP-2, a novel cysteine-rich bactericidal peptide from equine leukocytes.. Infect Immun 1992 Dec;60(12):5042-7.
- Couto MA, Harwig SS, Lehrer RI. Selective inhibition of microbial serine proteases by eNAP-2, an antimicrobial peptide from equine neutrophils.. Infect Immun 1993 Jul;61(7):2991-4.
- Flo TH, Smith KD, Sato S, Rodriguez DJ, Holmes MA, Strong RK, Akira S, Aderem A. Lipocalin 2 mediates an innate immune response to bacterial infection by sequestrating iron.. Nature 2004 Dec 16;432(7019):917-21.
- Farnaud S, Evans RW. Lactoferrin--a multifunctional protein with antimicrobial properties.. Mol Immunol 2003 Nov;40(7):395-405.
- Appelmelk BJ, An YQ, Geerts M, Thijs BG, de Boer HA, MacLaren DM, de Graaff J, Nuijens JH. Lactoferrin is a lipid A-binding protein.. Infect Immun 1994 Jun;62(6):2628-32.
- Roberts RM, Raub TJ, Bazer FW. Role of uteroferrin in transplacental iron transport in the pig.. Fed Proc 1986 Sep;45(10):2513-8.
- Wooding FB, Morgan G, Fowden AL, Allen WR. Separate sites and mechanisms for placental transport of calcium, iron and glucose in the equine placenta.. Placenta 2000 Sep;21(7):635-45.
- Stewart F, Kennedy MW, Suire S. A novel uterine lipocalin supporting pregnancy in equids.. Cell Mol Life Sci 2000 Sep;57(10):1373-8.
- Van den Steen PE, Proost P, Wuyts A, Van Damme J, Opdenakker G. Neutrophil gelatinase B potentiates interleukin-8 tenfold by aminoterminal processing, whereas it degrades CTAP-III, PF-4, and GRO-alpha and leaves RANTES and MCP-2 intact.. Blood 2000 Oct 15;96(8):2673-81.
- McQuibban GA, Gong JH, Wong JP, Wallace JL, Clark-Lewis I, Overall CM. Matrix metalloproteinase processing of monocyte chemoattractant proteins generates CC chemokine receptor antagonists with anti-inflammatory properties in vivo.. Blood 2002 Aug 15;100(4):1160-7.
- Mercer PF, Shute JK, Bhowmik A, Donaldson GC, Wedzicha JA, Warner JA. MMP-9, TIMP-1 and inflammatory cells in sputum from COPD patients during exacerbation.. Respir Res 2005 Dec 22;6(1):151.
- Kotilainen T, Huhtinen M, Katila T. Sperm-induced leukocytosis in the equine uterus.. Theriogenology 1994 Feb 2;41(3):629-36.
- Kenney R, Doig P. Equine endometrial biopsy. Current therapy in theriogenology: diagnosis, treatment, and prevention of reproductive diseases in small and large animals 1986;pp. 723–729.
- Christoffersen M, Woodward EM, Bojesen AM, Petersen MR, Squires EL, Lehn-Jensen H, Troedsson MH. Effect of immunomodulatory therapy on the endometrial inflammatory response to induced infectious endometritis in susceptible mares.. Theriogenology 2012 Sep 15;78(5):991-1004.
- Davis EG, Sang Y, Blecha F. Equine beta-defensin-1: full-length cDNA sequence and tissue expression.. Vet Immunol Immunopathol 2004 May;99(1-2):127-32.
- Ye J, Coulouris G, Zaretskaya I, Cutcutache I, Rozen S, Madden TL. Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction.. BMC Bioinformatics 2012 Jun 18;13:134.
- Traub-Dargatz JL, Salman MD, Voss JL. Medical problems of adult horses, as ranked by equine practitioners.. J Am Vet Med Assoc 1991 May 15;198(10):1745-7.
- Maloufi F, Pierson R, Otto S, Ball C, Card CR. Mares susceptible or resistant to endometritis have similar endometrial echographic and inflammatory cell reactions at 96 hours after infusion with frozen semen and extender. Proc Am Assoc Equine Pract 2002;48:51–57.
- Cui J, Zhu L, Xia X, Wang HY, Legras X, Hong J, Ji J, Shen P, Zheng S, Chen ZJ, Wang RF. NLRC5 negatively regulates the NF-kappaB and type I interferon signaling pathways.. Cell 2010 Apr 30;141(3):483-96.
- Davis BK, Roberts RA, Huang MT, Willingham SB, Conti BJ, Brickey WJ, Barker BR, Kwan M, Taxman DJ, Accavitti-Loper MA, Duncan JA, Ting JP. Cutting edge: NLRC5-dependent activation of the inflammasome.. J Immunol 2011 Feb 1;186(3):1333-7.
- Conti P, Boucher W, Letourneau R, Feliciani C, Reale M, Barbacane RC, Vlagopoulos P, Bruneau G, Thibault J, Theoharides TC. Monocyte chemotactic protein-1 provokes mast cell aggregation and [3H]5HT release.. Immunology 1995 Nov;86(3):434-40.
- Taub DD, Lloyd AR, Conlon K, Wang JM, Ortaldo JR, Harada A, Matsushima K, Kelvin DJ, Oppenheim JJ. Recombinant human interferon-inducible protein 10 is a chemoattractant for human monocytes and T lymphocytes and promotes T cell adhesion to endothelial cells.. J Exp Med 1993 Jun 1;177(6):1809-14.
- Cole KE, Strick CA, Paradis TJ, Ogborne KT, Loetscher M, Gladue RP, Lin W, Boyd JG, Moser B, Wood DE, Sahagan BG, Neote K. Interferon-inducible T cell alpha chemoattractant (I-TAC): a novel non-ELR CXC chemokine with potent activity on activated T cells through selective high affinity binding to CXCR3.. J Exp Med 1998 Jun 15;187(12):2009-21.
- Liao F, Rabin RL, Yannelli JR, Koniaris LG, Vanguri P, Farber JM. Human Mig chemokine: biochemical and functional characterization.. J Exp Med 1995 Nov 1;182(5):1301-14.
- Yung SC, Murphy PM. Antimicrobial chemokines.. Front Immunol 2012;3:276.
- Ulug U, Goldman S, Ben-Shlomo I, Shalev E. Matrix metalloproteinase (MMP)-2 and MMP-9 and their inhibitor, TIMP-1, in human term decidua and fetal membranes: the effect of prostaglandin F(2alpha) and indomethacin.. Mol Hum Reprod 2001 Dec;7(12):1187-93.
- Christiaens I, Zaragoza DB, Guilbert L, Robertson SA, Mitchell BF, Olson DM. Inflammatory processes in preterm and term parturition.. J Reprod Immunol 2008 Oct;79(1):50-7.
- Walter I, Handler J, Miller I, Aurich C. Matrix metalloproteinase 2 (MMP-2) and tissue transglutaminase (TG 2) are expressed in periglandular fibrosis in horse mares with endometrosis.. Histol Histopathol 2005 Oct;20(4):1105-13.
- Lehrer RI, Barton A, Daher KA, Harwig SS, Ganz T, Selsted ME. Interaction of human defensins with Escherichia coli. Mechanism of bactericidal activity.. J Clin Invest 1989 Aug;84(2):553-61.
- Pellegrini A, Waiblinger S, Von Fellenberg R. Purification of equine neutrophil lysozyme and its antibacterial activity against gram-positive and gram-negative bacteria.. Vet Res Commun 1991;15(6):427-35.
- Weinrauch Y, Elsbach P, Madsen LM, Foreman A, Weiss J. The potent anti-Staphylococcus aureus activity of a sterile rabbit inflammatory fluid is due to a 14-kD phospholipase A2.. J Clin Invest 1996 Jan 1;97(1):250-7.
- Hayes M, Quinn B, Lillie B, Côté O, Bienzle D, Waelchli R, Betteridge K. Changes in various endometrial proteins during cloprostenol-induced failure of early pregnancy in mares. Anim Reprod Sci 2012;9:723–741.
- Li C, Chan YR. Lipocalin 2 regulation and its complex role in inflammation and cancer.. Cytokine 2011 Nov;56(2):435-41.
Citations
This article has been cited 7 times.- Lindsay CV, Potter JA, Grimshaw AA, Abrahams VM, Tong M. Endometrial responses to bacterial and viral infection: a scoping review. Hum Reprod Update 2023 Sep 5;29(5):675-693.
- Jacobsen S, Mortensen CD, Høj EA, Vinther AM, Berg LC, Adler DMT, Verwilghen D, van Galen G. Neutrophil Gelatinase-Associated Lipocalin in Synovial Fluid from Horses with and without Septic Arthritis. Animals (Basel) 2022 Dec 21;13(1).
- Amaral A, Fernandes C, Rebordão MR, Szóstek-Mioduchowska A, Lukasik K, Pinto-Bravo P, Telo da Gama L, Jan Skarzynski D, Ferreira-Dias G. Myeloperoxidase Inhibition Decreases the Expression of Collagen and Metallopeptidase in Mare Endometria under In Vitro Conditions. Animals (Basel) 2021 Jan 16;11(1).
- Cassimeris L, Engiles JB, Galantino-Homer H. Interleukin-17A pathway target genes are upregulated in Equus caballus supporting limb laminitis. PLoS One 2020;15(12):e0232920.
- Crociati M, Capomaccio S, Mandara MT, Stradaioli G, Sylla L, Monaci M, Cappelli K. Different expression of Defensin-B gene in the endometrium of mares of different age during the breeding season. BMC Vet Res 2019 Dec 21;15(1):465.
- Wang JQ, Liu YR, Xia Q, Chen RN, Liang J, Xia QR, Li J. Emerging Roles for NLRC5 in Immune Diseases. Front Pharmacol 2019;10:1352.
- Holmes CM, Babasyan S, Wagner B. Neonatal and maternal upregulation of antileukoproteinase in horses. Front Immunol 2024;15:1395030.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists