{"paper_id":"ed068999-21b2-49f0-94b7-d86c8a7b0cc2","body_text":"1493\n© Aluna Publishing\n   Wiadomości Lekarskie Medical Advances, VOLUME LXXIX, ISSUE 7, JUL Y 2026\nINTRODUCTION\nAdenomyosis is a chronic disease of the female \nreproductive system that affects up to 60% of women \nof reproductive age worldwide [1]. Adenomyosis is \ncharacterised by benign invasion of the endometrium \ninto the myometrium [2]. Disruption of myometrium \nleads to pathology that affects female reproductive \nhealth and includes multiple clinical manifestations \nincluding excessive uterine bleeding, dysmenorrhea, \nchronic pelvic pain, and reduced fertility [3, 4]. Despite \nwell-defined clinical characteristics of adenomyosis, \nmolecular and cellular mechanisms that underlie the \ndisease origin and progression remain poorly under -\nstood. Determining the molecular basis of the develop-\nment of adenomyosis will help design better diagnostic \ntools and treatment regimens for women.\nA number of factors have been proposed to con-\ntribute to the development of adenomyosis including \nangiogenic abnormalities [5], hormonal imbalance \n[6], and genetic factors [7]. More recently, a number of \nstudies have revealed the importance of the immuno-\nlogical and pathogenic components in a related endo-\nmetrial condition endometriosis [8, 9]. A local immune \ndysregulation as well as changes in the microbiome in \nthe genital tract have been shown to play a role in the \ndisease development [10]. Infectious agents including \nbacteria, viruses and fungi can serve as a trigger to ini-\ntiate a dysbiosis in the genital tract leading to chronic \ninflammation and immune dysregulation [11, 12]. The \ninability of the immune system to resolve inflammation \nand clear the infection agents results in tissue damage \nand pathogenesis. Understanding the immunological \nnetworks of endometrial-myometrial pathology will \nbe invaluable in defining the biological markers of the \ndisease. \nThere are multiple lines of defence against the \nharmful factors that lead to the changes in the female \nreproductive flora and contribute to the disease pro -\ngression. The first line of defence includes cytokines \nand antimicrobial peptides (AMPs) released by immune \nThe analysis of cervicovaginal microbiota and immune \nmediators in patients with adenomyosis\nMykola Shcherbina, Liliya Potapova, Oksana Lipko, lrina Shcherbina, Anastasiia Chekhunova, Olga Mertsalova\nKHARKIV NATIONAL MEDICAL UNIVERSITY , KHARKIV , UKRAINE\nABSTRACT\nAim: To determine and characterise changes associated with microbial, inflammatory and immune factors in patients with adenomyosis.\nMaterials and Methods: 115 female patients were enrolled in the study, including 85 patients with adenomyosis and 30 healthy individuals. Vaginal \ncleanliness was assessed by collecting vaginal samples and plating on solid bacterial agar. Urogenital infections were determined by the PCR analysis of the \ncervicovaginal samples. The immunological characterisation was performed using enzyme immunoassay to analyse cervicovaginal lavage for β-defensin-2 \nand a defined set of cytokines. Antibody titers were quantified in patients’ serum using nephelometry.\nResults: Patients with adenomyosis displayed disrupted cervicovaginal microflora that was characterised by elevated levels of pro-inflammatory cytokines \nand antimicrobial peptide β-defensin-2. Serum analysis of patients with adenomyosis revealed higher concentration of IgG, IgM and IgA antibodies compared \nto healthy control groups.\nConclusions: Analysis of inflammatory and immunological parameters in patients with adenomyosis revealed distinct features of the pathophysiology of \nadenomyosis. Specifically, patients with adenomyosis had disrupted cervicovaginal microflora with higher prevalence of anaerobic species. Furthermore, \npatients with adenomyosis had elevated levels of pro-inflammatory cytokines and interleukins in cervicovaginal lavage, and higher levels of IgG, IgM and \nIgA antibody serum titres. The analysis presented in the paper represents an important step towards determining precise diagnostic parameters and effective \ntherapies to treat adenomyosis.\n  KEYWORDS: antimicrobial peptides, adenomyosis, cervicovaginal microbiome\nWiad Lek. 2026;79(7):1493-1499. doi: 10.36740/WLek/218754 DOI\nORIGINAL ARTICLE CONTENTS\n\nMykola Shcherbina et al. \n1494\ncells in response to microbial agent, inflammation \nand physical damage [13]. There are approximately \n39 antimicrobial peptides in cervicovaginal fluid [14] \nwith β-defensins family, cathelicidin  and SLPIs being \nthe most dominant AMPs [15]. Multiple reports have \ndemonstrated the importance of AMPs in the protec -\ntion of vaginal microbiome [16, 17]. However, the role \nof AMPs in adenomyosis remains poorly understood. \nRelease of AMPs, such as β-defensins, serves as a che -\nmoattractant for immune cells. Following AMPs release, \nmonocytes and dendritic cells migrate to the site of \ninfection and can phagocytose and present pathogen \nassociated molecular patters for T cell recognition. Upon \nactivation, T cells trigger the next line of defence known \nas the adaptive immunity that involves activation of B \ncells and production of different antibodies including \nIgG, IgM and IgA that act at mucosal barriers as well \nand tissues and blood [18].\nIn this work, we defined the components of innate \nand adaptive immunity in the cervicovaginal micro -\nenvironment in patients with adenomyosis. We show \nthat both microflora and immune composition is al-\ntered when adenomyosis is present, implicating these \nchanges in disease progression.  \nAIM\nTo determine and characterize changes associated with \nmicrobial, inflammatory and immune factors in patients \nwith adenomyosis.\nMATERIALS AND METHODS\nPATIENTS INVOLVED IN THE STUDY\n115 female patients aged 18-45 years old, who under-\nwent treatment at the clinical bases of the Department \nof Obstetrics and Gynecology No. 1 of the KhNMU, \ntook part in the study. The patients were assigned into \ngroups as follows. The first clinical group (study group) \nincluded 85 (73.9% of the total number of patients \nenrolled in the study) patients with adenomyosis. The \nsecond clinical group (control group) consisted of 30 \n(26.1% of the total number of patients enrolled in the \nstudy) healthy fertile women. All patients underwent a \nroutine general clinical examination during the course \nof the study, in accordance with the current clinical \nrecommendations of the Ministry of Health of Ukraine. \nThe examination was carried out after obtaining the \npatient’s informed consent for the diagnosis and pro-\ncessing of personal data.\nAdenomyosis was diagnosed in patients in the \nstudy group based on the diagnostic tools and \nhistological methods of examination of tissues ob -\ntained during hysteroscopy. Patient exclusion criteria \nincluded extragenital pathology, oncology includ -\ning leiomyomas, inflammation of the genitals, HPV \ninfection. Patients who were exposed to urogenital \nviral or bacterial infection in the last 6 months and \nwho underwent antibiotic therapy were excluded \nfrom the study.\nETHICS STATEMENT\nThe study was conducted in compliance with the basic \nbioethical norms and requirements of the Helsinki Dec-\nlaration of the General Assembly of the World Medical \nAssociation on Human Rights and Biomedicine (1977), \nin accordance with the provisions of the WHO, the In-\nternational Council of Medical Scientific Societies, the \nInternational Code of Medical Ethics (1983) and the \nOrder of the Ministry of Health of Ukraine No. 690 dated \nSeptember 23, 2009. The study protocol was approved \nby the Bioethics Commission of the KhNMU.\n[World Medical Association Declaration of Helsinki \n“Ethical Principles of Medical Research Involving Human \nSubjects” . 2008. URL : https://zakon.rada.gov.ua/laws/\nshow/990_005]\nDETECTION OF UROGENITAL INFECTIONS\nVaginal cleanliness was determined by isolating the \nLactobacilli strains from vaginal samples using standard \nprocedure of culturing bacteria on solid agar and by \nperforming subsequent PCR sequencing. \nTo determine the presence of anaerobic bacteria, cer-\nvicovaginal lavage (CVL) was analyzed for the presence \nof the indicated microbial agents (Table 1). Bacterial \nagents were detected by PCR using standard procedure. \nIMMUNOLOGICAL CHARACTERISATION\nCVL samples were collected and the levels of cytokines \nwere determined by solid-phase enzyme immunoassay \nmethod using Stat Enzyme Immunoassay Analyzer Fax \n® 303 Plus (Awareness Technology, Inc. USA ) according \nto manufacturer instructions.\nANALYSIS OF ANTIBODIES IN BLOOD\nBlood samples were collected during the study and \nprocessed immediately post-collection. Serum was \nseparated from whole blood samples by centrifugation \nat 2,000g for 15 min at 4 oC. Separated serum samples \nwere analysed for total immunoglobulins IgG, IgM and \nIgA using nephelometry.\n\nThe analysis of cervicovaginal microbiota and immune mediators in patients with adenomyosis\n1495\nSTATISTICAL ANALYSIS\nStatistical analysis was performed using software pack-\nage “Statistica 10.0 for Windows” . For each variation \nseries, we calculated average arithmetic mean (M), av-\nerage quadratic deviation (σ), average mistake average \narithmetic (m). To identify significant differences, Mann-\nWhitney U test was used. Differences were considered \nsignificant at a level of P < 0.05.\nRESUL TS \nANALYSIS OF BACTERIAL COMPOSITIONS \nIN THE CERVICOVAGINAL SECRETIONS IN \nPATIENTS WITH ADENOMYOSIS\nThere were 115 female patients involved in the study \nand the patients were divided into two groups. Group \n1 constituted patients with adenomyosis diagnosis \n(study group, 85 patients) and group 2 included healthy \npatients (control group, 30 patients). The patients were \n31.2 ± 2.7 years old, with no statistical difference in \nage  between the study or control groups. 30% of the \npatients in the study group contained cases of gyne -\ncological complications and family history of gynae -\ncological conditions compared to 10% in the control \ngroup. Average duration of adenomyosis in patients \nin study group was between 3 to 5 years. The clinical \nsigns of pathology in the study group included pelvic  \npain (71% of patients), dysmenorrhea (43% patients) \nand infertility (84% patients).\nFirst, we aimed to determine the degree of vaginal \ncleanliness among all the patients enrolled in the study. \nWe used previously reported lactobacilli grading meth-\nod [19] to define four degrees of purity of vaginal flora: \nGrade I defined as dominant lactobacilli with no other \nbacteria present, Grade II defined as having lactobacilli \nand other bacteria present, Grade II defined as few or \nno lactobacilli with other bacteria present and Grade \nIV defined as no lactobacilli and no other bacteria pres-\nent. Vaginal discharge was collected from patients and \nbacterial flora was examined using PCR (Table 1). Grade \nI vaginal purity was detected in 24 (80.0%) women \nof the control group and in 8 (9.4%) patients in the \nstudy group. Grade II-III vaginal purity was identified \nin 6 (20.0%) control group patients and in 71 patients \n(83.5%) of the study group. Grade IV vaginal purity was \npresent in 6 (7.1%) of cases in patients of the study \ngroup and was not detected in any of the patients in \nTable 1. Cervicovaginal cleanliness in the examined patients\nDegree of vaginal cleanliness\nStudy group (n=85) Control group (n=30)\nn % n %\nI 8 9.4 24 80\nII-III 71 83.5 6 20\nIV 6 7.1 - -\n% is percentage out of the total number of examined patients per group\nn-number of patients\nSource: compiled by the authors of this study\nTable 2. Concentration of antimicrobial peptide β-defensin-2 and cytokines in the cervicovaginal lavage of the examined patients. \nStudy group Control group\nn=85 n=30\nAntimicrobial peptide (AMP) ng /ml\nβ-defensin-2 30.5±3.5* 46.7±3.8\nCytokines pg /ml\nIL-4 20.0±1.8* 15.3±1.5\nIL-8 4.94±0.6* 3.8±0.4\nIL-10 8.37±0.2* 7.1±0.9\nIL-18 14.0±1.5* 10.9±1.3\nTNFα 43.2±4.5* 33.4±3.6\nVEGF 228.5±25.1* 173.5±16.9\nIFNg 9.54±1.1* 7.9±0.2\nMann-Whitney U test was performed. Mean±SEM is shown where SEM is standard error of the mean. *P<0.05 was considered statistically significant \nbetween the control and study groups. n is number of patients per group\nSource: compiled by the authors of this study\n\nMykola Shcherbina et al. \n1496\nThere different types of AMPs in the endometrial fluid \n[24] and β-defensins are of the most abundant AMPs \npresent. As a representative analysis, we, chose to \nlook at the levels of β-defensin-2 in CVL fluid in the \nstudy and control groups. The levels of b-defensins \nwere downregulated in patients with adenomyosis \ncompared to the control group (Table 2). This result \nsupported the link between adenomyosis and dysreg-\nulated microflora. \nCERVICOVAGINAL INFLAMMATORY \nCHANGES IN PATIENTS WITH ADENOMYOSIS\nWe hypothesised that disrupted microbial composition \nwithin the vaginal environment may also be accompa-\nnied by immunological changes. \nWe therefore examined the levels of the key immu-\nnological regulators including interleukins (IL-4, IL-8, \nIL-10, IL-18), pro-inflammatory mediators (TNFα, IFNγ) \nand endothelial growth factor (VEGF) in the CVL in the \nstudy and control groups.  In CVL, the concentration \nof IL-8, IL-10 and IL-18 was significantly higher in \npatients with adenomyosis compared to the control \ngroup (Table 2). Since IL-18 stimulates the formation \nof Th1-type cytokines (IFNγ, TNFα), its increase at the \nlocal level may be aimed at limiting inflammatory \nprocesses in the reproductive tract. In addition, by \nsecreting Th1-type mediators, IL-18 may contribute to \nthe development of a T-cell immune response. In line \nwith these observations, higher levels of TNFα were \ndetected in patients with adenomyosis compared to the \nstudy group (Table 2). IL-4 and IL-10, that are associated \nwith anti-inflammatory responses, were elevated in \npatients with adenomyosis. The concentration of VEGF \nwas significantly higher in the study group compared \nto healthy volunteers indicating a possible disruption \nin the normal angiogenesis that may result in vascular \ngrowth and proliferation of ectopic endometrium.\nNext, we examined the levels of three subclasses of \nantibodies IgG, IgA and IgM in the serum of patients of \nadenomyosis or healthy women. All three subclasses of \nantibodies were elevated in patients with adenomyosis \nthe control group. Together, these data indicated that \nvaginal purity is negatively affected in patients with \nadenomyosis. \nDecrease in vaginal purity suggested that microbial \nchanges may be present in the vaginal environment. \nPrevious work suggested that there is an association \nbetween anaerobic microbes and inflammation of en-\ndometrium [20, 21] which prompted us to ask whether \nanaerobic bacteria may play a role in adenomyosis. To \naddress this question, we examined the anaerobic mi-\ncrobial composition of the cervicovaginal flora in the \nstudy and control groups. Samples of cervicovaginal \nlavage were collected and microbial composition was \ndetermined by PCR for an indicated set of microbial \nagents (Fig. 1). Anaerobic microbial agents constitut -\ned seven microorganisms that have previously been \nreported in vaginal microflora [22, 23]. Comparison \nbetween the control and study groups revealed that \npatients with adenomyosis were characterized by the \npresence of a number of anaerobic microorganisms \nwith the Streptococcus spp. Staphylococcus spp. and  \nEubacterium spp. being second and third most abun-\ndant anaerobe present respectively. A single case of \nCandida spp., and another case of Mycoplasma hominis \nwere identified in the study group, suggesting that the \npresence of these microorganisms is rare yet detect -\nable. Unlike the study group, the vaginal microflora \nof healthy patients had no presence of any of the an-\naerobic microorganisms, consistent with the control \ngroup exhibiting Grade I degree with vaginal purity \n(Table 1). Together, these results suggested a possible \nlink between the adenomyosis and the presence of \nanaerobic species in likely disrupted cervicovaginal \nmicrobiota. \nDisruption in microflora in the vaginal environment \nin patients with adenomyosis lead us to examine the \nlocal immunological composition. AMPs produced \nby commensal microbiota are known to be the key \nmediators of defence against microbial pathogens \nin female reproductive tract [12]. AMPs are known to \nfluctuate in response to hormonal changes as well as \nin certain disease of female reproductive tract [13]. \nTable 3. Concentration of antibodies in the serum of the examined patients.\nStudy group Control group\nn=85 n=30\nIgM (g/l) 1.85 ±0.26* 1.02±0.09\nIgG (g/l) 16.6 ±2.5* 12.3 ±1.6\nIgA (g/l) 2.33 ±0.34* 1.10 ±0.08\nMann-Whitney U test was performed, Mean±SEM is shown where SEM is standard error of the mean. *P<0.05 was considered statistically significant \nbetween the control and study groups. n is number of patients per group\nSource: compiled by the authors of this study\n\nThe analysis of cervicovaginal microbiota and immune mediators in patients with adenomyosis\n1497\nof adenomyosis present themselves with defined \npatterns of cervicovaginal microbial composition and \nexpand our analysis to other AMPs in adenomyosis. \nThe striking difference in pro-inflammatory cytokine \nprofile between the patients with adenomyosis sug-\ngests that immune cell composition at the endometrial \nsite differs between the adenomyosis patients and \nthe healthy women. It would be interesting to define \nimmune subsets in the cervicovaginal microenviron-\nment in patients with adenomyosis to pinpoint the \ndominant immune cell populations that trigger the \nrelease of the proinflammatory cytokines identified \nin the present study. Interestingly, the levels of two \nanti-inflammatory cytokines IL-4 and IL-10 were sim-\nilarly elevated in patients with adenomyosis (Table \n2), indicating that even these elevated levels of an-\nti-inflammatory cytokines are insufficient to suppress \ninflammation at the lesion site. Future work will focus \non expanding the analysis of cytokine profiles to wider \nclinical groups to determine whether the cytokines \ncan be used as prognosis markers for the development \nof adenomyosis [26]. \nIdentifying the association between the elevated an-\ntibody levels and the presence of adenomyosis (Table \n3) raises an interesting possibility that adenomyosis \nmay have the autoimmune component its pathophys-\niology, similar to other endometriosis lesions [27]. The \nelevated levels of the antibodies in the serum of ade -\nnomyosis patients suggests that adaptive immunity \nmediated by T and B cells is activated in these patients. \ncompared to the control group, with IgG levels being \nthe most dominant class (Table 3). Collectively these \nresults indicated that the inflammatory microenviron-\nment, defined by elevated levels of pro-inflammatory \nmediators and increased inflammation-associated \nimmunoglobulins, may be a characteristic of adeno -\nmyosis lesions. \nDISCUSSION\nThe results of the study show that adenomyosis may \nbe accompanied by changes in the cervicovaginal mi-\ncrobiome, a drop in b-defensin-2 and changes in the \nlocal immunological profile. \nEpithelial cells of female reproductive tract are \ncolonized by commensal bacteria that secrete AMPs \namong others, thereby providing protection against \nharmful pathogens. Our results showed that patients \nwith adenomyosis displayed disruption in cervicovag-\ninal microbiome that was manifested by the presence \nof anaerobic microorganisms (Fig. 1, Table 1). Consis-\ntent with this observations, other reports indicated \nthat endometriosis-related lesions have distinct micro-\nbial compositions [11] and different stages of endome-\ntriosis are characterised by specific microbial patters \n[25]. In our study, we did not look at individual stages \nof adenomyosis. Furthermore, we only looked at the \nlevels of β-defensin-2, one of many AMPs secreted by \ncommensal bacteria in female genital tract. In future, \nwe would like to determine whether certain stages \nFig. 1. The presence of anaerobic microbial species in cervicovaginal lavage of patients with and without adenomyosis\nThe number of patients is presented as a percentage per either study (black) or control (grey) groups. Black bars depict patients in the study group \n(with adenomyosis, total n=84). No anaerobic bacteria were identified in the control group of patients (n=30) “X” indicates that no patients were \nidentified with a given microbial species. \nAuthors’ own work\n \nFig. 1. The presence of anaerobic microbial species in cervicovaginal lavage of patients with and \nwithout adenomyosis \n*The number of patients is presented as a percentage per either study (black) or control (grey) \ngroups. Black bars depict patients in the study group (with adenomyosis, total n=84). No anaerobic \nbacteria were identified in the control group of patients (n=30) “X” indicates that no patients were \nidentified with a given microbial species.  \nPicture taken by the authors \n \n\n\nMykola Shcherbina et al. \n1498\nCONCLUSIONS\nThe pathophysiology of adenomyosis remains understud-\nied largely due to the complexity of the diagnosis and \nlimited understanding of the etiology of the condition. \nThe study presented here revealed several important \naspects of the inflammation and immune dysregulation \nthat accompany adenomyosis lesions. The results showed \nthat patients with adenomyosis exhibit disrupted micro-\nbial composition within the cervicovaginal environment \ncharacterised by high prevalence of anaerobic species. \nCervicovaginal lavage collected from patients with adeno-\nmyosis was characterized by elevated levels of interleukins \n(IL-4, IL-8, IL-10, IL-18), pro-inflammatory mediators (TNFα, \nIFNy) and endothelial growth factor (VEGF). Furthermore, \nthe serum concentrations of IgG, IgM and IgA antibodies \nwere elevated in patients with adenomyosis compared to \nhealthy individuals. Our study provides an important foun-\ndation to further dissect the immunological composition of \nadenomyosis and ultimately help design better diagnostic \nand therapeutic tools to treat this debilitating condition. \nFollow up studies are necessary to determine whether \nthese antibodies are acting at the site of adenomyotic \nlesion, thereby exacerbating the disease further. An-\nother outstanding question remains - what are the \nantigens that are recognised by these antibodies in pa-\ntients with adenomyosis? Provided the elevated levels \nof antibodies in patients with adenomyosis recognize \nself-antigens at the adenomyosis lesion, it would be \ninteresting to investigate whether the mechanism \nof immunosuppression might be effective at certain \nstages of adenomyosis development. \nAdenomyosis remains to be a topical global problem \naffecting health of millions of women worldwide and \nthe current treatment does not address prevention or \npathophysiological aspects of the disease. The study \npresented here will contribute to a better under -\nstanding of the molecular mechanisms that govern \nadenomyosis at the cellular level and help advance \nthe development of much needed diagnostic and \nprevention strategies. \nREFERENCES\n 1.  Bourdon M, et al. Adenomyosis: An update regarding its diagnosis and clinical features. J Gynecol Obstet Hum Reprod. 2021;50(10):102228. \ndoi:10.1016/j.jogoh.2021.102228. DOI\n 2.  Bird CC, McElin TW, Manalo-Estrella P . The elusive adenomyosis of the uterus--revisited. Am J Obstet Gynecol. 1972;112(5):583-593. \ndoi:10.1016/0002-9378(72)90781-8. DOI\n 3.  Taylor MA, Croudace TJ, Muir FE, McBride M. Women’s experiences of living with adenomyosis and perceptions of the diagnostic journey: \na scoping review. BMJ Open. 2025;15(1):e087122. doi:10.1136/bmjopen-2024-087122. DOI\n 4.  eBioMedicine. Adenomyosis: an underacknowledged cause of abnormal uterine bleeding and pelvic pain. eBioMedicine. 2025;113:105656. \ndoi:10.1016/j.ebiom.2025.105656. DOI\n 5.  Harmsen MJ et al. Role of angiogenesis in adenomyosis-associated abnormal uterine bleeding and subfertility: a systematic review. \nHum Reprod Update. 2019;25(5):647-671. doi:10.1093/humupd/dmz024. DOI\n 6.  García-Solares J, Donnez J, Donnez O, Dolmans MM. Pathogenesis of uterine adenomyosis: invagination or metaplasia? Fertil Steril. \n2018;109(3):371-379. doi:10.1016/j.fertnstert.2017.12.030. DOI\n 7.  Bulun SE, Yildiz S, Adli M, Wei JJ. Adenomyosis pathogenesis: insights from next-generation sequencing. Hum Reprod Update. \n2021;27(6):1086-1097. doi:10.1093/humupd/dmab017. DOI\n 8.  Greene AD, Lang SA, Kendziorski JA et al. Endometriosis: where are we and where are we going? Reproduction. 2016;152(3):R63-R78. \ndoi:10.1530/REP-16-0052. DOI\n 9.  Anahtar MN et al. Cervicovaginal bacteria are a major modulator of host inflammatory responses in the female genital tract. Immunity. \n2015;42(5):965-976. doi:10.1016/j.immuni.2015.04.019. DOI\n 10.  Bourdon M et al. Immunological changes associated with adenomyosis: a systematic review. Hum Reprod Update. 2021;27(1):108-129. \ndoi:10.1093/humupd/dmaa038. DOI\n 11.  Muraoka A et al. Fusobacterium infection facilitates the development of endometriosis through the phenotypic transition of endometrial \nfibroblasts. Sci Transl Med. 2023;15(700):eadd1531. doi:10.1126/scitranslmed.add1531. DOI\n 12.  Gholiof M, Adamson-De Luca E, Wessels JM. The female reproductive tract microbiotas, inflammation, and gynecological conditions. \nFront Reprod Health. 2022;4:963752. doi:10.3389/frph.2022.963752. DOI\n 13.  Yarbrough VL, Winkle S, Herbst-Kralovetz MM. Antimicrobial peptides in the female reproductive tract: a critical component of the mucosal \nimmune barrier with physiological and clinical implications. Hum Reprod Update. 2015;21(3):353-377. doi:10.1093/humupd/dmu065. DOI\n 14.  Muytjens CMJ, Yu Y , Diamandis EP . Discovery of antimicrobial peptides in cervical-vaginal fluid from healthy nonpregnant women via \nan integrated proteome and peptidome analysis. Proteomics. 2017;17(21):1600461. doi:10.1002/pmic.201600461. DOI\n 15.  Madanchi H et al. Expression of cathelicidin, ERK, MyD88, and TLR-9 in the blood of women in the pre-pregnancy, pregnancy, and their \ninfant cord blood. Hum Immunol. 2022;83(12):826-831. doi:10.1016/j.humimm.2022.08.014. DOI\n\nThe analysis of cervicovaginal microbiota and immune mediators in patients with adenomyosis\n1499\n 16.  Agarwal N, Buckley N, Nakra N et al. Activity of genital tract secretions and synthetic antimicrobial peptides against Group B Streptococcus. \nAm J Reprod Immunol. 2015;74(6):499-507. doi:10.1111/aji.12427. DOI\n 17.  Doerflinger SY , Throop AL, Herbst-Kralovetz MM. Bacteria in the vaginal microbiome alter the innate immune response and barrier \nproperties of the human vaginal epithelia in a species-specific manner. J Infect Dis. 2014;209(12):1989-1999. doi:10.1093/infdis/jiu004. \nDOI\n 18.  Chen K, Magri G, Grasset EK, Cerutti A. Rethinking mucosal antibody responses: IgM, IgG and IgD join IgA. Nat Rev Immunol. 2020;20(7):427-\n441. doi:10.1038/s41577-019-0261-1. DOI\n 19.  Falagas ME, Betsi GI, Athanasiou S. Probiotics for the treatment of women with bacterial vaginosis. Clin Microbiol Infect. 2007;13(7):657-\n664. doi:10.1111/j.1469-0691.2007.01688.x. DOI\n 20.  Haggerty CL, Hillier SL, Bass DC, Ness RB. Bacterial vaginosis and anaerobic bacteria are associated with endometritis. Clin Infect Dis. \n2004;39(7):990-995. doi:10.1086/423963. DOI\n 21.  Leonardi M et al. Transvaginal ultrasound can accurately predict the American Society of Reproductive Medicine stage of endometriosis \nassigned at laparoscopy. J Minim Invasive Gynecol. 2020;27(7):1581-1587.e1. doi:10.1016/j.jmig.2020.02.014. DOI\n 22.  Larsen B, Monif GRG. Understanding the bacterial flora of the female genital tract. Clin Infect Dis. 2001;32(4):e69-e77. doi:10.1086/318710. \nDOI\n 23.  Takimoto K, Yamada H, Shimada S et al. Chronic endometritis and uterine endometrium microbiota in recurrent implantation failure \nand recurrent pregnancy loss. Biomedicines. 2023;11(9):2391. doi:10.3390/biomedicines11092391. DOI\n 24.  Azkargorta M et al. In-depth proteomics and natural peptidomics analyses reveal antibacterial peptides in human endometrial fluid. J \nProteomics. 2020;216:103652. doi:10.1016/j.jprot.2020.103652. DOI\n 25.  Cregger M et al. Reproductive microbiomes: using the microbiome as a novel diagnostic tool for endometriosis. Reprod Immunol Open \nAccess. 2017;2(3):36. doi:10.21767/2476-1974.100036. DOI\n 26.  Dechaud H, Haouzi D, Vincens C et al. New biomarkers of adenomyosis in endometrium. Fertil Steril. 2014;102(3):e49-e50. doi:10.1016/j.\nfertnstert.2014.07.172. DOI\n 27.  Gleicher N, el-Roeiy A, Confino E, Friberg J. Is endometriosis an autoimmune disease? Obstet Gynecol. 1987;70(1):115-122. \ndoi:10.1016/0020-7292(88)90292-5. DOI\nCONFLICT OF INTEREST\nThe Authors declare no conflict of interest\nCORRESPONDING AUTHOR\nOlga Mertsalova\nKharkiv National Medical University\n4 Nauky Avenue, 61022, Kharkiv, Ukraine\ne-mail: merts@ukr.net\nORCID AND CONTRIBUTIONSHIP\nMykola Shcherbina: 0000-0003-3605-7204  \nLiliya Potapova: 0000-0002-2022-5333  \nOksana Lipko: 0000-0001-6462-3563  \nlrina Shcherbina: 0000-0001-7666-8953  \nAnastasiia Chekhunova: 0000-0002-6509-5747  \nOlga Mertsalova: 0000-0001-9967-4918  \n – Work concept and design,  – Data collection and analysis,  – Responsibility for statistical analysis,  – Writing the article,  – Critical review,  – Final approval of the article\nRECEIVED: 07.11.2025\nACCEPTED: 02.03.2026\n CREATIVE COMMONS 4.0","source_license":"CC0","license_restricted":false}