Populations of NK Cells and Regulatory T Cells in the Endometrium of Cycling Mares-A Preliminary Study.
Abstract: Endometrial immune cells are essential to support uterine functions across the estrous cycle and in preparation for pregnancy. It has been acknowledged that changes in phenotype and/or numbers of lymphocytes, such as regulatory T cells (Tregs) and NK cells, might result in lower fertility in women and mice. Little is known about equine endometrial immune cells across the estrous cycle. Here, we compared the populations of endometrial Tregs and NK cells in estrus and diestrus in mares. Endometrial biopsy and blood samples were taken in estrus and diestrus from 11 mares ages 4-12 years. Flow cytometry with anti-CD4, -CD25 and -FOXP3 and anti-NKp46 and -CD3 antibodies was used to determine the populations of Tregs and NK cells, respectively. The concentration of progesterone was measured with chemiluminescence immunoassay. The results were analyzed with paired Student tests. The mean percentage of endometrial CD4FOXP3 Tregs was 13.7 ± 6.2% in diestrus and 14.5 ± 5.9% in estrus, while the mean percentage of endometrial CD4FOXP3CD25 Tregs changed from 3.6 ± 2.1% in diestrus to 2 ± 2% in estrus ( = 0.0947). The mean proportion of CD3NKp46 lymphocytes in the endometrium was not significantly different, with 6 ± 1% in estrus and 6.5 ± 1.4% in diestrus. There was a large variation in the percentage of NK cells between mares of 2.1-12.7%. This study showed, for the first time, the presence of CD4FOXP3CD25 Tregs and CD3NKp46 NK cells in the endometrium of non-pregnant cycling mares. The percentage of Tregs, and to a greater extent NK cells, showed large fluctuations between mares. Both Tregs and NK cells might be important for the preparation of the endometrium for semen deposition and pregnancy; however, further research is required.
Publication Date: 2022-11-30 PubMed ID: 36496894PubMed Central: PMC9740928DOI: 10.3390/ani12233373Google Scholar: Lookup
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- Journal Article
Summary
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This research study explores the populations of certain immune cells (regulatory T cells and NK cells) in the endometrium of mares during different stages of their reproductive cycle. The study ascertains the presence of these cells and their potential role in fertility, foundational knowledge for future research looking at improving fertility in mares.
Objective of the Study
- The study aimed to compare the populations of regulatory T cells (Tregs) and NK cells in the endometrium of mares during both the estrus and diestrus phases of the estrous cycle.
Study Design and Participants
- Endometrial biopsy and blood samples were taken from 11 mares of varying ages (between 4 and 12 years) during both their estrus and diestrus phases.
Methodology
- The researchers utilized a lab technique known as flow cytometry, along with specific antibodies targeting CD4, CD25, FOXP3, NKp46, and CD3 markers, to identify and quantify the populations of Tregs and NK cells in the samples.
- The team also measured the concentration of progesterone, a hormone that plays an important role in the mare’s reproductive cycle, using a chemiluminescence immunoassay.
- The results were then statistically analysed using paired Student t-tests.
Findings of the Study
- The study demonstrated for the first time the presence of CD4FOXP3CD25 Tregs and CD3NKp46 NK cells in the endometrium of non-pregnant, cycling mares.
- The proportions of these cells varied significantly amongst individual mares, particularly the NK cells, with quantities ranging between 2.1% and 12.7%.
- There was no significant difference in the proportion of these cells between the estrus and diestrus phases of the reproductive cycle.
Implications of the Study
- The study suggests that both Tregs and NK cells potentially play important roles in preparing the endometrium for semen deposition and pregnancy in mares.
- The presence of these cells and their variations in mare endometrium adds to the limited existing knowledge and provides grounds for further research, particularly in determining their roles in the fertility of mares.
Cite This Article
APA
Jaworska J, de Mestre AM, Wiśniewska J, Wagner B, Nowicki A, Kowalczyk-Zięba I, Wocławek-Potocka I.
(2022).
Populations of NK Cells and Regulatory T Cells in the Endometrium of Cycling Mares-A Preliminary Study.
Animals (Basel), 12(23), 3373.
https://doi.org/10.3390/ani12233373 Publication
Researcher Affiliations
- Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-747 Olsztyn, Poland.
- Department of Comparative Biomedical Sciences, Royal Veterinary College, Hertfordshire, Hatfield AL9 7TA, UK.
- Laboratory of Cell and Tissue Analysis and Imaging, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-747 Olsztyn, Poland.
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
- Private Veterinary Practice Arkadiusz Nowicki, 10-684 Olsztyn, Poland.
- Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-747 Olsztyn, Poland.
- Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-747 Olsztyn, Poland.
Grant Funding
- NCN grant 2020/36/C/NZ9/00192 / National Science Center
Conflict of Interest Statement
The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.
References
This article includes 38 references
- Lee SK, Kim CJ, Kim DJ, Kang JH. Immune cells in the female reproductive tract.. Immune Netw 2015 Feb;15(1):16-26.
- Berbic M, Fraser IS. Immunology of normal and abnormal menstruation.. Womens Health (Lond) 2013 Jul;9(4):387-95.
- Croy BA, Wessels J, Linton N, Tayade C. Comparison of immune cell recruitment and function in endometrium during development of epitheliochorial (pig) and hemochorial (mouse and human) placentas.. Placenta 2009 Mar;30 Suppl A:S26-31.
- Manaster I, Mandelboim O. The unique properties of uterine NK cells.. Am J Reprod Immunol 2010 Jun;63(6):434-44.
- Kofod L, Lindhard A, Hviid TVF. Implications of uterine NK cells and regulatory T cells in the endometrium of infertile women.. Hum Immunol 2018 Sep;79(9):693-701.
- Watson ED, Thomson SR. Lymphocyte subsets in the endometrium of genitally normal mares and mares susceptible to endometritis.. Equine Vet J 1996 Mar;28(2):106-10.
- Tunon AM, Katila T, Magnusson U, Nummijarvi A, Rodriguez-Martinez H. Distribution of T cells in the endometrium of the mare 6 and 48 h after insemination.. Reprod. Dom. Anim. 2002;34:443–444.
- Noronha LE, Huggler KE, de Mestre AM, Miller DC, Antczak DF. Molecular evidence for natural killer-like cells in equine endometrial cups.. Placenta 2012 May;33(5):379-86.
- de Mestre A, Noronha L, Wagner B, Antczak DF. Split immunological tolerance to trophoblast.. Int J Dev Biol 2010;54(2-3):445-55.
- Feuerer M, Hill JA, Mathis D, Benoist C. Foxp3+ regulatory T cells: differentiation, specification, subphenotypes.. Nat Immunol 2009 Jul;10(7):689-95.
- Sakaguchi S, Miyara M, Costantino CM, Hafler DA. FOXP3+ regulatory T cells in the human immune system.. Nat Rev Immunol 2010 Jul;10(7):490-500.
- Robbin MG, Wagner B, Noronha LE, Antczak DF, de Mestre AM. Subpopulations of equine blood lymphocytes expressing regulatory T cell markers.. Vet Immunol Immunopathol 2011 Mar 15;140(1-2):90-101.
- Kallikourdis M, Betz AG. Periodic accumulation of regulatory T cells in the uterus: preparation for the implantation of a semi-allogeneic fetus?. PLoS One 2007 Apr 18;2(4):e382.
- de Mestre AM, Rose BV, Chang YM, Wathes DC, Verheyen KLP. Multivariable analysis to determine risk factors associated with early pregnancy loss in thoroughbred broodmares.. Theriogenology 2019 Jan 15;124:18-23.
- Hamstead L, Chang YM, Crowhurst J, Wise Z, Mcgladdery A, Ricketts S, De Mestre AM. Retrospective study of early pregnancy loss in Thoroughbred mares.. Equine Vet. J. 2012;44:2–18.
- Noronha LE, Harman RM, Wagner B, Antczak DF. Generation and characterization of monoclonal antibodies to equine NKp46.. Vet Immunol Immunopathol 2012 Jun 15;147(1-2):60-8.
- Viveiros MM, Antczak DF. Characterization of equine natural killer and IL-2 stimulated lymphokine activated killer cell populations.. Dev Comp Immunol 1999 Sep;23(6):521-32.
- Blanchard-Channell M, Moore PF, Stott JL. Characterization of monoclonal antibodies specific for equine homologues of CD3 and CD5.. Immunology 1994 Aug;82(4):548-54.
- Aurich C. Reproductive cycles of horses.. Anim Reprod Sci 2011 Apr;124(3-4):220-8.
- Manaster I, Mizrahi S, Goldman-Wohl D, Sela HY, Stern-Ginossar N, Lankry D, Gruda R, Hurwitz A, Bdolah Y, Haimov-Kochman R, Yagel S, Mandelboim O. Endometrial NK cells are special immature cells that await pregnancy.. J Immunol 2008 Aug 1;181(3):1869-76.
- El Costa H, Tabiasco J, Berrebi A, Parant O, Aguerre-Girr M, Piccinni MP, Le Bouteiller P. Effector functions of human decidual NK cells in healthy early pregnancy are dependent on the specific engagement of natural cytotoxicity receptors.. J Reprod Immunol 2009 Nov;82(2):142-7.
- El Costa H, Casemayou A, Aguerre-Girr M, Rabot M, Berrebi A, Parant O, Clouet-Delannoy M, Lombardelli L, Jabrane-Ferrat N, Rukavina D, Bensussan A, Piccinni MP, Le Bouteiller P, Tabiasco J. Critical and differential roles of NKp46- and NKp30-activating receptors expressed by uterine NK cells in early pregnancy.. J Immunol 2008 Sep 1;181(5):3009-17.
- Liu S, Diao L, Huang C, Li Y, Zeng Y, Kwak-Kim JYH. The role of decidual immune cells on human pregnancy.. J Reprod Immunol 2017 Nov;124:44-53.
- Oladunni FS, Horohov DW, Chambers TM. EHV-1: A Constant Threat to the Horse Industry.. Front Microbiol 2019;10:2668.
- Robertson SA, Care AS, Moldenhauer LM. Regulatory T cells in embryo implantation and the immune response to pregnancy.. J Clin Invest 2018 Oct 1;128(10):4224-4235.
- Hartmann C, Gerner W, Walter I, Saalmüller A, Aurich C. Influences of intrauterine semen administration on regulatory T lymphocytes in the oestrous mare (Equus caballus).. Theriogenology 2018 Sep 15;118:119-125.
- Polanczyk MJ, Carson BD, Subramanian S, Afentoulis M, Vandenbark AA, Ziegler SF, Offner H. Cutting edge: estrogen drives expansion of the CD4+CD25+ regulatory T cell compartment.. J Immunol 2004 Aug 15;173(4):2227-30.
- Arruvito L, Sanz M, Banham AH, Fainboim L. Expansion of CD4+CD25+and FOXP3+ regulatory T cells during the follicular phase of the menstrual cycle: implications for human reproduction.. J Immunol 2007 Feb 15;178(4):2572-8.
- Guerin LR, Prins JR, Robertson SA. Regulatory T-cells and immune tolerance in pregnancy: a new target for infertility treatment?. Hum Reprod Update 2009 Sep-Oct;15(5):517-35.
- Hamza E, Gerber V, Steinbach F, Marti E. Equine CD4(+) CD25(high) T cells exhibit regulatory activity by close contact and cytokine-dependent mechanisms in vitro.. Immunology 2011 Nov;134(3):292-304.
- Aurich C, Weber J, Nagel C, Merkl M, Jude R, Wostmann S, Ollech D, Baron U, Olek S, Jansen T. Low levels of naturally occurring regulatory T lymphocytes in blood of mares with early pregnancy loss.. Reprod Fertil Dev 2014;26(6):827-33.
- Zenclussen ML, Thuere C, Ahmad N, Wafula PO, Fest S, Teles A, Leber A, Casalis PA, Bechmann I, Priller J, Volk HD, Zenclussen AC. The persistence of paternal antigens in the maternal body is involved in regulatory T-cell expansion and fetal-maternal tolerance in murine pregnancy.. Am J Reprod Immunol 2010 Mar 1;63(3):200-8.
- Granne I, Shen M, Rodriguez-Caro H, Chadha G, O'Donnell E, Brosens JJ, Quenby S, Child T, Southcombe JH. Characterisation of peri-implantation endometrial Treg and identification of an altered phenotype in recurrent pregnancy loss.. Mucosal Immunol 2022 Jan;15(1):120-129.
- Parham P, Moffett A. Variable NK cell receptors and their MHC class I ligands in immunity, reproduction and human evolution.. Nat Rev Immunol 2013 Feb;13(2):133-44.
- Maher JK, Tresnan DB, Deacon S, Hannah L, Antczak DF. Analysis of MHC class I expression in equine trophoblast cells using in situ hybridization.. Placenta 1996 Jul-Aug;17(5-6):351-9.
- Futas J, Oppelt J, Janova E, Musilova P, Horin P. Complex variation in the KLRA (LY49) immunity-related genomic region in horses.. HLA 2020 Sep;96(3):257-267.
- Todd ET, Thomson PC, Hamilton NA, Ang RA, Lindgren G, Viklund Å, Eriksson S, Mikko S, Strand E, Velie BD. A genome-wide scan for candidate lethal variants in Thoroughbred horses.. Sci Rep 2020 Aug 4;10(1):13153.
- Chazara O, Xiong S, Moffett A. Maternal KIR and fetal HLA-C: a fine balance.. J Leukoc Biol 2011 Oct;90(4):703-16.
Citations
This article has been cited 1 times.- Satué K, La Fauci D, Medica P, Damiá Gímenez E, Cravana C, Fazio E. Shifts between pro-inflammatory and anti-inflammatory profiles in pregnant mares: a review of physiological functions. Front Vet Sci 2025;12:1660759.
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