[en] Adenomyosis-the ectopic presence of endometrial glands and stroma within the myometrium-affects reproductive-age women and is associated with pain, bleeding, and subfertility, yet the immune events that precede pregnancy remain poorly defined. To address this, we investigated how the uterine immune microenvironment evolves before conception in a tamoxifen-induced murine model of adenomyosis. At 1- and 3-months post-induction, we analyzed uterine tissue by immunofluorescence, flow cytometry, and quantitative polymerase chain reaction. We found that adenomyotic uteri exhibited a sustained elevation of interleukin-6 messenger RNA, a transient interleukin-10 rise at 1 month, and stable cyclooxygenase-2 levels. Interleukin-6 receptor messenger RNA and signal transducer and activator of transcription 3 messenger RNA were both transiently downregulated at 1 month and returned to control levels by 3 months. Early in disease development, total macrophage numbers declined and displayed an alternative (M2) activation phenotype, followed by a selective loss of classically activated (M1) macrophages at later stages. B lymphocytes were consistently enriched, indicating enhanced humoral activity. Although overall T-cell counts remained stable, the CD3+ compartment underwent a marked shift from double-negative T lymphocytes toward T-helper and cytotoxic subsets, coinciding with the transient signaling changes. Limitations of our study include reliance on a single animal model, analysis at only two timepoints, and a lack of functional assessment of regulatory T cells. Future work should incorporate finer temporal profiling, single-cell transcriptomics, and validation in human tissues. These findings highlight dynamic innate-adaptive crosstalk as an early driver of adenomyosis pathology and suggest that targeting interleukin-6-mediated pathways may inform biomarker development and novel immunomodulatory interventions.
Disciplines :
Reproductive medicine (gynecology, andrology, obstetrics)
Vervier, Julie ; Université de Liège - ULiège > GIGA > GIGA Cancer - Tumors Biology and Development
Bernet, Laëtitia ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques
Camboni, Alessandra ; Gynecology Research Unit, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium ; Anatomopathology Department, Cliniques Universitaires Saint-Luc, Brussels, Belgium
Dolmans, Marie-Madeleine; Gynecology Research Unit, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium ; Gynecology Department, Cliniques Universitaires Saint-Luc, Brussels, Belgium
Munaut, Carine ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques
Language :
English
Title :
Temporal dynamics of uterine immune microenvironment remodeling in a murine model of adenomyosis.
Bacci M, Capobianco A, Monno A, Cottone L, Di Puppo F, Camisa B, Mariani M, Brignole C, Ponzoni M, Ferrari S et al. Macrophages are alternatively activated in patients with endometriosis and required for growth and vascularization of lesions in a mouse model of disease. Am J Pathol 2009; 175: 547-556.
Barrier BF, Malinowski MJ, Dick EJ, Hubbard GB, Wright Bates G. Adenomyosis in the baboon is associated with primary infertility. Fertil Steril 2004; 82: 1091-1094.
Bergeron C, Amant F, Ferenczy A. Pathology and physiopathology of adenomyosis. Best Pract Res Clin Obstet Gynaecol 2006; 20: 511-521.
Bird CC, McElin TW, Manalo-Estrella P. The elusive adenomyosis of the uterus-revisited. Am J Obstet Gynecol 1972; 112: 583-593.
Bourdon M, Maget AS, Jeljeli M, Doridot L, Marcellin L, Thomas M, Chene C, Chouzenoux S, Batteux F, Chapron C et al. Reduced fertility in adenomyosis mouse model is associated with an altered immune profile in the uterus during the implantation period. Hum Reprod 2024; 39: 119-129.
Bourdon M, Mimouni A, Maignien C, Casalechi M, Vigano P, Bordonne C, Millisher AE, Patrat C, Marcellin L, Chapron C et al. Reduced live birth rates following ART in adenomyosis patients: a matched control study. Hum Reprod 2025; 40: 855-864.
Bourdon M, Santulli P, Doridot L, Jeljeli M, Chêne C, Chouzenoux S, Nicco C, Marcellin L, Chapron C, Batteux F. Immune cells and Notch1 signaling appear to drive the epithelial to mesenchymal transition in the development of adenomyosis in mice. Mol Hum Reprod 2021; 27: gaab053.
Bourdon M, Santulli P, Jeljeli M, Vannuccini S, Marcellin L, Doridot L, Petraglia F, Batteux F, Chapron C. Immunological changes associated with adenomyosis: a systematic review. Hum Reprod Update 2021; 27: 108-129.
Campo S, Campo V, Benagiano G. Adenomyosis and infertility. Reprod Biomed Online 2012; 24: 35-46.
Cao Y, Yang D, Cai S, Yang L, Yu S, Geng Q, Mo M, Li W, Wei Y, Li Y et al. Adenomyosis-associated infertility: an update of the immunological perspective. Reprod Biomed Online 2025; 50: 104703.
Chapron C, Tosti C, Marcellin L, Bourdon M, Lafay-Pillet MC, Millischer AE, Streuli I, Borghese B, Petraglia F, Santulli P. Relationship between the magnetic resonance imaging appearance of adenomyosis and endometriosis phenotypes. Hum Reprod 2017; 32: 1393-1401.
Chen Y, Guo Y, Li S, Xu J, Zhao C, Wang J, Yang J, Ning W, Qu Y, Zhang M et al. Tumor-derived IL-6 promotes chordoma invasion by stimulating tumor-associated macrophages M2 polarization and TNFα secretion. Int Immunopharmacol 2024; 143: 113315.
De Clercq K, Hennes A, Vriens J. Isolation of mouse endometrial epithelial and stromal cells for in vitro decidualization. JoVE 2017; 2017: 55168
Couper KN, Blount DG, Riley EM. IL-10: the master regulator of immunity to infection. J Immunol 2008; 180: 5771-5777.
El-Zayadi AA, Mohamed SA, Arafa M, Mohammed SM, Zayed A, Abdelhafez MS, Badawy AM. Anti-IL-6 receptor monoclonal antibody as a new treatment of endometriosis. Immunol Res 2020; 68: 389-397.
Gao Y, Zhao H, Wang P, Wang J, Zou L. The roles of SOCS3 and STAT3 in bacterial infection and inflammatory diseases. Scand J Immunol 2018; 88: e12727.
García-Solares J, Donnez J, Donnez O, Dolmans MM. Pathogenesis of uterine adenomyosis: invagination or metaplasia? Fertil Steril 2018; 109: 371-379.
Gordon S. Alternative activation of macrophages. Nat Rev Immunol 2003; 3: 23-35.
Green AR, Styles JA, Parrott EL, Gray D, Edwards RE, Smith AG, Gant TW, Greaves P, Al-Azzawi F, White INH. Neonatal tamoxifen treatment of mice leads to adenomyosis but not uterine cancer. Exp Toxicol Pathol 2005; 56: 255-263.
Guo S, Li Z, Yan L, Sun Y, Feng Y. GnRH agonist improves pregnancy outcome in mice with induced adenomyosis by restoring endometrial receptivity. Drug Des Devel Ther 2018; 12: 1621-1631.
Hagen M, Pangrazzi L, Rocamora-Reverte L, Weinberger B. Legend or Truth: Mature CD4+CD8+ Double-Positive T Cells in the Periphery in Health and Disease. Biomedicines 2023; 11: 2702.
Incognito GG, Di Guardo F, Gulino FA, Genovese F, Benvenuto D, Lello C, Palumbo M. Interleukin-6 as a useful predictor of endometriosis-associated infertility: a systematic review. Int J Fertil Steril 2023; 17: 226-230.
Itoh F, Komohara Y, Takaishi K, Honda R, Tashiro H, Kyo S, Katabuchi H, Takeya M. Possible involvement of signal transducer and activator of transcription-3 in cell-cell interactions of peritoneal macrophages and endometrial stromal cells in human endometriosis. Fertil Steril 2013; 99: 1705-1713.
Jin Z, Wu X, Liu H, Xu C. Celecoxib, a selective COX-2 inhibitor, markedly reduced the severity of tamoxifen-induced adenomyosis in a murine model. Exp Ther Med 2020; 19: 3289-3299.
Kisovar A, Becker CM, Granne I, Southcombe JH. The role of CD8+T cells in endometriosis: a systematic review. Front Immunol 2023; 14: 1225639.
Leary S, Underwood W, Anthony R, Cartner S, Grandin T, Greenacre C, Gwaltney-Brant S, McCrackin MA, Meyer R, Miller D et al. AVMA guidelines for the euthanasia of animals: 2020 edition. Members of the Panel on Euthanasia AVMA Staff Consultants. 2020. Available from: https://www.avma.org/sites/default/files/2020-02/Guidelines-on-Euthanasia-2020.pdf
Li J, Wang R, Chen J, Xu A, Fu Y, Lin Y, Wang X, Ye S, Yuan Y, Du F et al. Efficacy and safety of ruxolitinib in adult patients with refractory rheumatic disease-associated macrophage activation syndrome. Front Immunol 2025; 16: 1604648.
Li Q, Shi J, Yi D, Li X, Gu Z, Yan H, Leng J. The pathogenesis of endometriosis and adenomyosis: insights from single-cell RNA sequencing. Biol Reprod 2024; 110: 854-865.
Lin PF, Lan XL, Chen FL, Yang YZ, Jin YP, Wang AH. Reference gene selection for real-time quantitative PCR analysis of the mouse uterus in the peri-implantation period. PLoS One 2013; 8: e62462.
Liu W, Sheng S, Zhu C, Li C, Zou Y, Yang C, Chen ZJ, Wang F, Jiao X. Increased NKG2A+CD8+T-cell exhaustion in patients with adenomyosis. Mucosal Immunol 2023; 16: 121-134.
Mao Q, Liang XL, Wu YF, Pang YH, Zhao XJ, Lu YX. ILK promotes survival and self-renewal of hypoxic MSCs via the activation of lncTCF7-Wnt pathway induced by IL-6/STAT3 signaling. Gene Ther 2019; 26: 165-176.
Marquardt RM, Jeong JW, Fazleabas AT. Animal models of adenomyosis. Semin Reprod Med 2020; 38: 168-178.
Martinez FO, Gordon S. The M1 and M2 paradigm of macrophage activation: time for reassessment. F1000Prime Rep 2014; 6: 13.
Moore KW, De Waal Malefyt R, Coffman RL, O'Garra A. Interleukin-10 and the interleukin-10 receptor. Annu Rev Immunol 2001; 19: 683-765.
Nie MF, Xie Q, Wu YH, He H, Zou LJ, She XL, Wu XQ. Serum and ectopic endometrium from women with endometriosis modulate macrophage M1/M2 polarization via the Smad2/Smad3 pathway. J Immunol Res 2018; 2018: 6285813.
Ota H, Igarashi S, Hatazawa J, Tanaka T. Is adenomyosis an immune disease? Hum Reprod Update 1998; 4: 360-367.
Parel Y, Chizzolini C. CD4+CD8+double positive (DP) T cells in health and disease. Autoimmun Rev 2004; 3: 215-220.
Parisi S, Ditto MC, Ghellere F, Panaro S, Piccione F, Borrelli R, Fusaro E. Update on tocilizumab in rheumatoid arthritis: a narrative review. Front Immunol 2025; 16: 1470488.
Park W, Lim W, Kim M, Jang H, Park SJ, Song G, Park S. Female reproductive disease, endometriosis: from inflammation to infertility. Mol Cells 2025; 48: 100164.
Parrott E, Butterworth M, Green A, White IN, Greaves P. Adenomyosis-a result of disordered stromal differentiation. Am J Pathol 2001; 159: 623-630.
Pinzauti S, Lazzeri L, Tosti C, Centini G, Orlandini C, Luisi S, Zupi E, Exacoustos C, Petraglia F. Transvaginal sonographic features of diffuse adenomyosis in 18-30-year-old nulligravid women without endometriosis: association with symptoms. Ultrasound Obstet Gynecol 2015; 46: 730-736.
Riccio LdGC, Santulli P, Marcellin L, Abrão MS, Batteux F, Chapron C. Immunology of endometriosis. Best Pract Res Clin Obstet Gynaecol 2018; 50: 39-49.
Squatrito M, Vervier J, Bindels J, Bernet L, Blacher S, Nisolle M, Munaut C. Impaired fertility in adenomyosis: a murine model reveals endometrial receptivity and progesterone resistance imbalances. Reproduction 2024; 167: e240019.
Stratopoulou CA, Cussac S, d'Argent M, Donnez J, Dolmans M-M. M2 macrophages enhance endometrial cell invasiveness by promoting collective cell migration in uterine adenomyosis. Reprod Biomed Online 2023; 46: 729-738.
Stratopoulou CA, Donnez J, Dolmans MM. Origin and pathogenic mechanisms of uterine adenomyosis: what is known so far. Reprod Sci 2021; 28: 2087-2097.
Takebayashi A, Kimura F, Kishi Y, Ishida M, Takahashi A, Yamanaka A, Wu D, Zheng L, Takahashi K, Suginami H et al. Subpopulations of macrophages within eutopic endometrium of endometriosis patients. Am J Reprod Immunol 2015; 73: 221-231.
Tanaka T, Narazaki M, Kishimoto T. IL-6 in inflammation, immunity, and disease. Cold Spring Harb Perspect Biol 2014; 6: a016295.
Taskin MI, Gungor AC, Adali E, Yay A, Onder GO, Inceboz U. A humanized anti-interleukin 6 receptor monoclonal antibody, tocilizumab, for the treatment of endometriosis in a rat model. Reprod Sci 2016; 23: 662-669.
Vannuccini S, Petraglia F. Recent advances in understanding and managing adenomyosis. F1000Res 2019; 8: 283.
Vercellini P, Buffo C, Bandini V, Cipriani S, Chiaffarino F, Viganò P, Somigliana E. Prevalence of adenomyosis in symptomatic adolescents and young women: a systematic review and meta-analysis. F&S Rev 2025; 6: 2666-5719.
Wang J, Huang C, Jiang R, Du Y, Zhou J, Jiang Y, Yan Q, Xing J, Hou X, Zhou J et al. Decreased endometrial IL-10 impairs endometrial receptivity by downregulating HOXA10 expression in women with adenomyosis. Biomed Res Int 2018; 2018: 2549789.
West NR. Coordination of immune-stroma crosstalk by IL-6 family cytokines. Front Immunol 2019; 10: 1093.
Yang HL, Zhou WJ, Chang KK, Mei J, Huang LQ, Wang MY, Meng Y, Ha SY, Li DJ, Li MQ. The crosstalk between endometrial stromal cells and macrophages impairs cytotoxicity of NK cells in endometriosis by secreting IL-10 and TGF-β. Reproduction 2017; 154: 815-825.
Zhihong N, Yun F, Pinggui Z, Sulian Z, Zhang A. Cytokine profiling in the eutopic endometrium of adenomyosis during the implantation window after ovarian stimulation. Reprod Sci 2016; 23: 124-133.
