Targeted transcript analysis revealed association of suboptimal expression of certain endometrial immunity-related genes with disparate uterine diseases in zebu cows

The onset of uterine infection during postpartum period compromises uterine health, fertility, and productivity of dairy cattle. Endometrial innate immunity plays a key role in eliminating uterine infection and keeping the uterus healthy. Hence, the present study has been designed with the hypothesis that altered endometrial immune response around calving may compromise uterine health during postpartum period. Expression of interleukins (IL-1β, IL-6, IL-8, IL-10, and TNF-α), prostaglandin synthase (PGFS, PGES), and antimicrobial peptides (beta-defensins (BDEF-4, BDEF-5), lingual antimicrobial peptide (LAP), and calcium-binding proteins (S100A8, S100A9, and S100A12) in endometrial tissues on the day of calving was studied using qRT-PCR, and circulating concentrations of prostaglandin E and F metabolites (PGEM and PGFM) during peripartum period (on days − 7, − 4, − 1 (before calving), 0 (on the day of calving), + 1, + 4, and + 7 (post calving)) of normal (healthy) cows (n = 11) that did not develop postpartum uterine infection and cows that developed puerperal metritis (n = 7) and clinical endometritis (n = 6) were studied. Endometrial expression of IL-1β, TNF-α, BDEF-4, BDEF-5, S100A8, S100A12, and PGFS was higher (P < 0.05), and expression of IL-6, IL-8, IL-10, and PGES was lower (P < 0.05) in normal (healthy) cows than puerperal metritic and clinical endometritic cows. The PGFM concentration in serum was high (P < 0.05) on days 0, + 1, and + 4 of calving in puerperal metritic cows followed by normal and clinical endometritic cows. However, PGEM concentration in serum was high (P < 0.05) during peripartum period in uterine-infected (puerperal metritic and clinical endometritic) cows compared with normal cows. From the above findings, it is concluded that higher constitutive expression of IL-1β, TNF-α, PGFS, BDEF-4, BDEF-5, S100A8, and S100A12 genes in the endometrium and lower concentration of PGEM during the period immediate to calving might be beneficial for uterine health of cows. Similar content being viewed by others

Akira, S. and Takeda, K., 2004. Toll-like receptor signalling, Nature Reviews Immunology, 4, 499–511. Baithalu, R.K., Singh, S.K., Kumaresan, A., Mohanty, A.K., Mohanty, T.K., Kumar, S., Kerketta, S., Maharana, B.R., Patbandha, T.K., Attupuram, N. and Agarwal, S.K., 2016. Transcriptional abundance of antioxidant enzymes in endometrium and their circulating levels in Zebu cows with and without uterine infection, Animal Reproduction Science, 177, 79–87. Bartlett, P.C., Kirk, J.H., Wilke, M.A., Kaneene, J.B. and Mather, E.C., 1986. Metritis complex in Michigan Holstein-Friesian cattle: incidence, descriptive epidemiology and estimated economic impact, Preventive Veterinary Medicine, 4, 235–48. Baumann, H. and Gauldie, J., 1994. The acute phase response. Immunol, Today 15 (2), 74–80. Beutler, B., 2004. Inferences, questions and possibilities in Toll-like receptor signalling, Nature 430, 257–263. Caraher, E.M., Gumulapurapu, K., Taggart, C.C., Murphy, P., McClean, S. and Callaghan, M., 2007. The effect of recombinant human lactoferrin on growth and the antibiotic susceptibility of the cystic fibrosis pathogen Burkholderiacepacia complex when cultured planktonically or as biofilms, Journal of Antimicroiology Chemotherapy, 60, 546–554. Chapwanya, A., Meade, K.G., Doherty, M.L., Callanan, J.J., Mee, J.F. and O’Farrelly, C., 2009. Histopathological and molecular evaluation of Holstein–Friesian cows postpartum: toward an improved understanding of uterine innate immunity, Theriogenology 71, 1396–1407. Chapwanya, A., Meade, K.G., Foley, C., Narciandi, F., Evans, ACO., Doherty, M.L., Callanan, J.J. and O’Farrelly, C., 2012. The postpartum endometrial inflammatory response: a normal physiological event with potential implications for bovine fertility, Reproduction Fertility Development, 24, 1028–1039. Cormican, P., Meade, K.G., Cahalane, S., Narciandi, F., Chapwanya, A., Lloyd, A.T. and O’Farrelly, C., 2008. Evolution, expression and effectiveness in a cluster of novel bovine beta-defensins, Immunogenetics, 60, 147–156. Davies, D., Meade, K.G., Herath, S., Eckersall, P.D., Gonzalez, D., White, J.O., Conlan, R.S., O’Farrelly, C. and Sheldon, I.M., 2008. Toll-like receptor and antimicrobial peptide expression in the bovine endometrium, Reproductive Biology and Endocrinology, 6, 53. 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, Reproduction Fertility Development, 22, 818–829. Gabler, C., Fischer, C., Drillich, M., Einspanier, R. and Heuwieser, W., 2010. Time-dependent mRNA expression of selected proinflammatory factors in the endometrium of primiparous cows postpartum, Reproductive Biology and Endocrinology, 8, 152. Galvao, K.N., Santosa, N.R., Galvao, J.S. and Gilbert, R.O., 2011. Association between endometritis and endometrial cytokine expression in postpartum Holstein cows, Theriogenology 76(2), 290–9. Hancock, R. E. and Scott, M. G. 2000. The role of antimicrobial peptides in animal defences. Proceedings of the National Academy of Sciences, USA 97, 8856–8861. Havelock, J.C., Keller, P., Muleba, N., Mayhew, B.A., Casey, B.M., Rainey, W.E. and Word, R.A., 2005. Human myometrial gene expression before and during parturition, Biology of Reproduction, 72, 707–719. Herath, S., Fischer, D.P., Werling, D., Williams, E.J., Lilly, S.T., Dobson, H., Bryant, C.E. and Sheldon, I.M., 2006. Expression and function of Toll-like receptor 4 in the endometrial cells of the uterus, Endocrinology, 147, 562–570. Herath, S., Lilly, S.T., Santos, N.R., Gilbert, R.O., Goetze, L., Bryant, C.E., White, J.O., Cronin, J. and Sheldon, I.M., 2009. Expression of genes associated with immunity in the endometrium of cattle with disparate postpartum uterine disease and fertility, Reproductive Biology and Endocrinology, 7, 55. Hoedemaker, M., Lund, L.A. and Wagner, W.C., 1992. Influence of arachidonic acid metabolites and steroids on function of bovine polymorphonuclear neutrophils, American Journal of Veterinary Research, 53, 1534–1539. Konigsson, K., Gustafsson, H. and Kindahl, H., 2002. 15-Ketodihydro-PGF2α, progesterone and uterine involution in primiparous cows with induced retained placenta and postpartal endometritis treated with oxytetracycline and flunixin. Reproduction in Domestic Animals, 37, 43–51. Lehrer, R.I., 2004. Primate defensins, Nature Reviews of Microbiology, 2, 727–738. Leslie, K.E., 1983. The Events of Normal and Abnormal Postpartum Reproductive Endocrinology and Uterine Involution in Dairy Cows: A Review, Canadian Veterinary Journal, 24, 67–71. Li, Q., Verma, I.M., 2002. NF-kappa B regulation in the immune system. Nature Reviews Immunology, 2, 725–734. Lindell, J.O., Kindahl, H., Jansson, I. and Edqvist, I.E. 1982. Post-partum release of prostaglandin F2a and Uterine involution in the cow, Theriogenology, 17(3), 237–245. Liu, S., Zuo, Z., Yang, Z., Liu, B. and Zhang, N., 2012. Roles of toll-like receptor 4 and immunomodulatory hormones in bovine endometritis, African Journal of Pharmacy and Pharmacology, 6 (15), 1136–1141. Livak, K.J. and Schmittgen, T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods, 25(4), 402–8. Loyi, T., Kumar, H., Nandi, S., Mathapati, B. S., Patra, M. K. and Pattnaik, B., 2013. Differential expression of pro-inflammatory cytokines in endometrial tissue of buffaloes with clinical and sub-clinical endometritis. Research in Veterinary Science, 94, 336–340. Martins, T.M., da Paixao, T.A., Costa, E.A., Pires, A.C., Santos, R.L. and Borges, A.M., 2011. Postpartum toll-like receptors and β-defensin 5 mRNA levels in the endometrium of Holstein cows. Veterinary Immunology and Immunopathology, 139, 277–281. Mateus, L., Lope de Costa, L., Carvalho, H., Serra, P. Silvia, J., 2003. Blood and intrauterine leucocyte profile and function in dairy cows that spontaneously recovered from postpartum endometritis. Reproduction in Domestic Animals, 37, 176–180. Mehrzad, J., Duchateau, L., Pyorola, S. and Burvenich, C., 2004. Blood and milk neutrophil chemiluminescence and viability in primiparous and pluriparous dairy cows during late pregnancy, around parturition and early lactation. Journal of Dairy Science, 85, 3268–3276. Mette, C., Dooleweerdt, B.C., Stine, J., Miki, B.A., Roenn, P.M. and Henrik, L.J., 2010. 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, Veterinary Immunology and Immunopathology, 138, 95–105. Mijatovic, T., Kruys, V., Caput, D., Defrance, P. and Huez, G., 1997. Interleukin-4 and -13 inhibit tumor necrosis factor-α mRNA translational activation in lipopolysaccharide-induced mouse macrophages, Journal of Biological Chemistry, 272, 14394–8. Mitchell, G.B., Albright, B.N. and Caswell, J. L., 2003. Effect of interleukin-8 and granulocyte colony-stimulating factor on priming and activation of bovine neutrophils. Infection and Immunity,71, 1643–1649. Nakao, T., Gamal, A., Osawa, T., Nakada, K., Moriyoshi, M. and Kawata, K., 1997. Postpartum plasma PGF metabolite profile in cows with dystocia and/or retained placenta, and effect of fenprostalene on uterine involution and reproductive performance. Journal of Veterinary Medical Science, 59(9), 791–794. NRC., 2001. National Research Council Nutrient Requirements of Dairy Cattle (Seventh revised edition). The National Academies Press, Washington, DC. Sahl, H.G., Pag, U., Bonness, S., Wagner, S., Antcheva, N. and Tossi, A., 2005. Mammalian defensins: structures and mechanism of antibiotic activity, Journal of Leukocyte Biology, 77, 466–475. Seals, R.C., Matomoros, I. and Lewis, G.S., 2002. Relationship between postpartum changes in 13, 14-dihydro-15-ketoprostaglandin F2α concentrations in Holstein cows and their susceptibility to endometritis, Journal of Animal Science, 80, 1068–1073. Sheldon, I.M., 2004. The postpartum uterus, Veterinary Clinics of North America: Food Animal Practice, 20, 569–591. Sheldon, I.M., Cronin, J., Goetze, L., Donofrio, G. and Schuberth, H.J., 2009. Defining postpartum uterine disease and the mechanisms of infection and immunity in the female reproductive tract in cattle, Biology of Reproduction, 81, 1025–1032. Slama, H., Vaillancourt, D. and Goff, A.K., 1991. Pathophysiology of puerperal period: relationship between prostaglandin E2 (PGE2) and uterine involution in the cow, Theriogenology, 36, 1071–1090. Sohnle, P.G., Haunter, M.J., Hahn, B., Chazin, W.J., 2000. Zinc-reversible antimicrobial activity of recombinant calprotectin (migration inhibitory factor-related proteins 8 and 14). The Journal of Infectious Diseases, 182 (4), 1272–5. Swangchan-Uthai, T., Lavender, C.R., Cheng, Z., Fouladi-Nashta, A.A. and Wathes, D.C., 2012. Time course of defense mechanisms in bovine endometrium in response to lipopolysaccharide, Biology of Reproduction, 87, 1–13. Tagashira, Y., Taniguchi, F., Harada, T., Ikeda, A., Watanabe, A. and Terakawa, N., 2008. Interleukin-10 attenuates TNF-alpha-induced interleukin-6 production in endometriotic stromal cells, Fertility and Sterility, 91, 2185–92. Wathes, D.C., Cheng, Z., Chowdhury, W., Fenwick, M.A., Fitzpatrick, R., Morris, D.G., Patton, J. and Murphy, J.J., 2009. Negative energy balance alters global gene expression and immune responses in the uterus of postpartum dairy cows, Physiological Genomics, 39, 1–13. Zerbe, H., Ossadnik, C., Leibold, W. and Schuberth, H. J., 2002. Lochial secretions of Escherichia coli- or Arcanobacterium pyogenes-infected bovine uteri modulate the phenotype and the functional capacity of neutrophilic granulocytes, Theriogenology, 57, 1161–1177. Acknowledgments The authors thank the Indian Council of Agricultural Research, Government of India, the Director, ICAR-National Dairy Research Institute, Karnal, Haryana, and the Director, ICAR-Indian Veterinary Research Institute, Bareilly, U.P., India, for providing necessary facilities and logistics support to conduct this study. Funding This study was funded by the National Dairy Research Institute, India. Author information Authors and Affiliations Corresponding authors Ethics declarations The procedures applied have been approved by the Institute Animal Ethics Committee (1705/Go/ac/13/CPCSEA). Conflict of interest The authors declare that they have no conflict of interest. Additional information Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Electronic supplementary material ESM 1 (download DOCX ) (DOCX 10 kb). Rights and permissions About this article Cite this article Baithalu, R.K., Singh, S.K., Kumaresan, A. et al. Targeted transcript analysis revealed association of suboptimal expression of certain endometrial immunity-related genes with disparate uterine diseases in zebu cows. Trop Anim Health Prod 51, 2493–2503 (2019). https://doi.org/10.1007/s11250-019-01958-3 Received: Accepted: Published: Version of record: Issue date: DOI: https://doi.org/10.1007/s11250-019-01958-3