{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T22:22:39Z","timestamp":1768947759164,"version":"3.49.0"},"reference-count":45,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T00:00:00Z","timestamp":1768521600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004329","name":"Slovenian Research and Innovation Agency","doi-asserted-by":"publisher","award":["P3-0327"],"award-info":[{"award-number":["P3-0327"]}],"id":[{"id":"10.13039\/501100004329","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Regulatory T cells (Tregs), particularly their phenotypically distinct subpopulations, are critical for the establishment of maternal immune tolerance during embryo implantation. Despite advances in assisted reproductive technologies, implantation failure remains a frequent and often unexplained clinical challenge. Variations in Treg frequency and phenotype have been proposed to influence implantation success, particularly under differing hormonal conditions. This study aimed to investigate peripheral blood Treg levels and their subpopulations on the day of blastocyst transfer in both stimulated in vitro fertilization (IVF\/ICSI) cycles involving controlled ovarian hyperstimulation (COH) and true natural cycles with frozen embryo transfer (FET), and to examine their associations with systemic hormone levels and anti-M\u00fcllerian hormone (AMH). A prospective observational study was conducted including women undergoing IVF\/ICSI with fresh embryo transfer (ET) and women undergoing natural cycle FET. Peripheral blood samples were collected on the day of ET and analyzed using 13-colour flow cytometry, enabling detailed subdivision of Tregs into multiple subpopulations based on the expression of differentiation and chemokine markers, including CXCR5. In addition, because common \u03b3-chain cytokines may influence pregnancy success by modulating the balance between suppressive Treg and non-Treg subsets, intracellular STAT5 signaling was assessed using phospho-specific flow cytometry. Serum estradiol, progesterone, FSH, LH, and AMH levels were measured in parallel. Significant differences were observed in Treg subpopulation distributions between women who conceived and those who did not. Higher frequencies of na\u00efve CXCR5\u2212 Tregs were associated with clinical pregnancy, independent of age, and correlated with serum progesterone levels. Moreover, both na\u00efve Treg frequency and enhanced IL-7-dependent STAT5 signaling in na\u00efve Tregs from women undergoing COH were associated with AMH levels, suggesting a link between ovarian reserve and Treg homeostasis mediated by signal transducer and activator of transcription 5 (STAT5) signaling. In conclusion, Treg subpopulations, particularly CXCR5\u2212 na\u00efve Tregs, appear to play a central role in implantation success following ET. Their distribution differs between stimulated and natural cycles and is influenced by systemic progesterone levels and STAT5 signaling. These findings suggest that peripheral Treg profiling may represent a potential biomarker of implantation competence and could inform personalized approaches in assisted reproduction.<\/jats:p>","DOI":"10.3390\/ijms27020922","type":"journal-article","created":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T09:24:34Z","timestamp":1768901074000},"page":"922","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Circulating Na\u00efve Regulatory T Cell Subset Displaying Increased STAT5 Phosphorylation During Controlled Ovarian Hyperstimulation Is Associated with Clinical Pregnancy and Progesterone Levels"],"prefix":"10.3390","volume":"27","author":[{"given":"Ksenija","family":"Raki\u0107","sequence":"first","affiliation":[{"name":"Department of Reproductive Medicine and Gynaecological Endocrinology, Clinic for Gynaecology and Perinatology, University Medical Centre Maribor, Ljubljanska ulica 5, 2000 Maribor, Slovenia"},{"name":"Faculty of Medicine, University of Maribor, Taborska ulica 8, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2188-578X","authenticated-orcid":false,"given":"Ale\u0161","family":"Goropev\u0161ek","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, University of Maribor, Taborska ulica 8, 2000 Maribor, Slovenia"},{"name":"Department of Laboratory Diagnostics, University Medical Centre Maribor, Ljubljanska ulica 5, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0934-5024","authenticated-orcid":false,"given":"Nejc","family":"Kozar","sequence":"additional","affiliation":[{"name":"Department of Reproductive Medicine and Gynaecological Endocrinology, Clinic for Gynaecology and Perinatology, University Medical Centre Maribor, Ljubljanska ulica 5, 2000 Maribor, Slovenia"},{"name":"Faculty of Medicine, University of Maribor, Taborska ulica 8, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3911-2408","authenticated-orcid":false,"given":"Borut","family":"Kova\u010di\u010d","sequence":"additional","affiliation":[{"name":"Department of Reproductive Medicine and Gynaecological Endocrinology, Clinic for Gynaecology and Perinatology, University Medical Centre Maribor, Ljubljanska ulica 5, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sara","family":"\u010curi\u010d","sequence":"additional","affiliation":[{"name":"Department of Laboratory Diagnostics, University Medical Centre Maribor, Ljubljanska ulica 5, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andreja","family":"Zakel\u0161ek","sequence":"additional","affiliation":[{"name":"Department of Laboratory Diagnostics, University Medical Centre Maribor, Ljubljanska ulica 5, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3904-1814","authenticated-orcid":false,"given":"Evgenija","family":"Hom\u0161ak","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, University of Maribor, Taborska ulica 8, 2000 Maribor, Slovenia"},{"name":"Department of Laboratory Diagnostics, University Medical Centre Maribor, Ljubljanska ulica 5, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0210-9467","authenticated-orcid":false,"given":"Milan","family":"Relji\u010d","sequence":"additional","affiliation":[{"name":"Department of Reproductive Medicine and Gynaecological Endocrinology, Clinic for Gynaecology and Perinatology, University Medical Centre Maribor, Ljubljanska ulica 5, 2000 Maribor, Slovenia"},{"name":"Faculty of Medicine, University of Maribor, Taborska ulica 8, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"570","DOI":"10.1093\/humupd\/dmad010","article-title":"Opening the black box: Why do euploid blastocysts fail to implant? 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