RESUMO
Endometriosis's pathophysiology remains incompletely understood, with evidence pointing towards a dysregulated immune response. Regulatory T (Treg) cells, pivotal in maintaining self-tolerance, may facilitate the survival of ectopic endometrial cells within the abdominal cavity, thereby contributing to endometriosis development. This study aimed to assess the prevalence of CD39+CD73+ suppressor Treg cell subsets in the peripheral blood of endometriosis patients. This research focuses on the pivotal role of regulatory T-cells (Tregs), which are essential for maintaining immune tolerance and preventing autoimmune diseases. A case-control study was conducted, including 32 women diagnosed with endometriosis and 22 control subjects. The frequency of peripheral blood CD39+CD73+ suppressor Treg cells was quantified using flow cytometry. No significant differences were observed in the frequency of CD3+CD4+CD25High cells (Median [M]: 10.1; Interquartile Range [IQR]: 6.32â18.3 vs. M: 9.72; IQR: 6.22-19.8) or CD3+CD4+CD25HighCD39+Foxp3+ cells (M: 31.1; IQR: 19.7-44.0 vs. M: 30.55; IQR: 18.5-45.5) between controls and patients. However, a significantly lower frequency of CD3+CD4+CD25HighCD39+CD73+ cells was observed in the endometriosis group compared to controls (M: 1.98; IQR: 0.0377-3.17 vs. M: 2.25; IQR: 0.50-4.08; p = 0.0483), suggesting a reduction in systemic immune tolerance among these patients. This finding highlights the potential role of CD39 and CD73 expression on Treg cells as biomarkers for assessing disease severity and progression. Furthermore, elucidating the mechanisms driving these alterations may unveil new therapeutic strategies to restore immune equilibrium and mitigate endometriosis symptoms.
Assuntos
Apirase , Endometriose , Citometria de Fluxo , Fatores de Transcrição Forkhead , Linfócitos T Reguladores , Humanos , Feminino , Endometriose/imunologia , Endometriose/sangue , Linfócitos T Reguladores/imunologia , Adulto , Estudos de Casos e Controles , Fatores de Transcrição Forkhead/sangue , Fatores de Transcrição Forkhead/análise , Apirase/análise , 5'-Nucleotidase/sangue , Adulto Jovem , Antígenos CD/sangue , Antígenos CD/análise , Estatísticas não Paramétricas , Valores de ReferênciaRESUMO
Endometriosis (EDT) is an inflammatory disease characterized by implantation/growth of endometrial tissue, glands, and/or stroma, outside the uterus. Reduced NK cell cytotoxic activity has been implicated in its pathogenesis, together with other immunologic alterations. We investigated the influence of KIR gene polymorphisms and their HLA ligand combinations in deep endometriosis (DE) susceptibility. One hundred sixty women with a histological diagnosis of DE and 202 control women without the disease, who underwent laparoscopy, were enrolled. The DE group was subdivided into initial (I/II; n = 60) and advanced stages (III/IV, n = 100). KIR and HLA class I gene polymorphisms were typed by PCR-SSP and sequence-based-typing (SBT), respectively. We observed a significant association of KIR2DL2, an inhibitory gene of B haplotype, conferring risk for DE in Euro-descendants. Positive associations of Bx haplotype and centromeric AB segments were also found. However, no association with KIR-HLA ligand combination was observed. Our data suggest KIR2DL2 gene to be a relevant factor favoring NK inhibition in DE in Euro-descendants, contributing to the defective NK cytotoxic activity and impaired clearance of ectopic endometrial cells in the disease.
Assuntos
Endometriose/genética , Polimorfismo de Nucleotídeo Único , Receptores KIR2DL2/genética , Adulto , Brasil/epidemiologia , Estudos de Casos e Controles , Endometriose/diagnóstico , Endometriose/etnologia , Endometriose/imunologia , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Haplótipos , Humanos , Células Matadoras Naturais/imunologia , Pessoa de Meia-Idade , Fenótipo , Medição de Risco , Fatores de Risco , Índice de Gravidade de Doença , População Branca/genética , Adulto JovemRESUMO
Background: Endometriosis does not have a well-established physiopathology. It has been addressed that endometriosis is an inflammatory disease, where endocrine-immunological interactions are probably involved in the pathogenesis of the disease. The role of the immune system in endometriosis has been suggested to play an important role in both initiation and progression of the disease.Methods: A search for the following keywords was performed in the PubMed database: "endometriosis", "endometriosis and ovarian cancer", "endometriosis and immunology", and "endometriosis and cytokines".Results: The articles identified were published in English between 1921 and 2020. We selected 100 articles for further analysis.Conclusion: The recognition of the direct involvement of these two important physiological mechanisms causes a change in the pathophysiological focus of the disease. Researching the activities of numerous cells involved in immune reactions may offer new therapeutic targets.
Assuntos
Endometriose/imunologia , Endometriose/epidemiologia , Endometriose/genética , Feminino , Humanos , Neoplasias Ovarianas/epidemiologia , Neoplasias Ovarianas/genéticaRESUMO
OBJECTIVE: To analyze and compare the expression of Toll-like receptors by regulatory T cells present in the peritoneal fluid of patients with and without endometriosis. METHODS: Regulatory T cells were isolated from peritoneal fluid of women with and without endometriosis, collected during surgery, and mRNA was extracted for analysis of Toll-like receptors expression by reverse-transcriptase polymerase chain reaction. RESULTS: Patients with endometriosis presented regulatory T cells expressing a larger number and variety of Toll-like receptors when compared to regulatory T cells from patients in the Control Group. Toll-like receptor-1 and Toll-like receptor-2 in regulatory T cells were expressed in both groups. All other expressed Toll-like receptors types were only found in regulatory T cells from the Endometriosis Group. CONCLUSION: Patients with endometriosis had peritoneal regulatory T cells expressing various Toll-like receptors types.
Assuntos
Líquido Ascítico/patologia , Endometriose/patologia , Endométrio/patologia , Linfócitos T Reguladores/química , Receptores Toll-Like/análise , Adolescente , Adulto , Líquido Ascítico/imunologia , Índice de Massa Corporal , Estudos de Casos e Controles , Endometriose/imunologia , Endométrio/imunologia , Feminino , Humanos , Valores de Referência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estatísticas não Paramétricas , Linfócitos T Reguladores/imunologia , Escala Visual Analógica , Adulto JovemRESUMO
Eutopic endometrium appears to be crucial for endometriosis development. Despite of the evident importance, data regarding the cellular microenvironment remain unclear. Our objective was to explore the tissue microenvironment heterogeneity, transcripts, and pathways that are enriched in all phases of the menstrual cycle by analysing publicly deposited data derived from whole transcriptome microarrays of eutopic endometria of women with and without endometriosis. A meta-analysis of the transcriptome microarrays was performed using raw data available from a public database. Eligibility criteria included eutopic endometrium samples from women with endometriosis and healthy controls without any pathological condition reported the presence of an adequately reported normal menstrual phase, and samples containing both glandular and stromal components. Raw data were processed using a robust multiarray average method to provide background correction, normalisation, and summarisation. The batch effect was estimated by principal variant component analysis and removed using an empirical Bayes method. Cellular tissue heterogeneity was inferred using the xCell package. Differentially expressed genes were identified based on a 5% adjusted p value and a 2.0-fold change. Pathways were identified by functional enrichment based on the Molecular Signatures Database, a p value of < 5%, and an FDR q value of ≤ 25%. Genes that were more frequently found in pathways were identified using leading edge analysis. In a manner independent of cycle phase, the subpopulations of activated dendritic cells, CD4 T effector memory phenotype cells, eosinophils, macrophages M1, and natural killer T cells (NKT) were all higher in stage I-II endometriosis compared to those in healthy controls. The subpopulations of M2 macrophages and natural killer T cells were elevated in eutopic endometriums from women with stage III-IV endometriosis, and smooth muscle cells were always more prevalent in healthy eutopic endometriums. Among the differently expressed genes, FOS, FOSB, JUNB, and EGR1 were the most frequently mapped within the interaction networks, and this was independent of stage and cycle phase. The enriched pathways were directly related to immune surveillance, stem cell self-renewal, and epithelial mesenchymal transition. PI3K AKT mTOR, TGF signalling, and interferon alpha/gamma responses were enriched exclusively in stage III-IV endometriosis. The cellular microenvironments and immune cell profiles were different between eutopic endometriums from women with stage I-II and stage III-IV endometriosis, and these differences were independent of the hormonal milieu. Specifically, a pro-inflammatory profile was predominant in stage I-II endometriosis, and M1-M2 polarization into eutopic endometrium may be crucial for the progression of the disease. The higher prevalence of NKT cells in eutopic endometriums from women with endometriosis that was independent of cycle phase or staging suggested a sustained stress and/or damage to these eutopic endometriums. Based on this, the results of this meta-analysis are important for identifying challenges and opportunities for future research.
Assuntos
Endometriose/patologia , Endométrio/metabolismo , Transcriptoma , Adulto , Estudos de Casos e Controles , Microambiente Celular , Análise por Conglomerados , Bases de Dados Factuais , Endometriose/imunologia , Endometriose/metabolismo , Transição Epitelial-Mesenquimal , Feminino , Perfilação da Expressão Gênica/métodos , Humanos , Fosfatidilinositol 3-Quinases/metabolismo , Análise de Componente Principal , Índice de Gravidade de Doença , Transdução de Sinais/genética , Serina-Treonina Quinases TOR/metabolismoRESUMO
ABSTRACT Objective To analyze and compare the expression of Toll-like receptors by regulatory T cells present in the peritoneal fluid of patients with and without endometriosis. Methods Regulatory T cells were isolated from peritoneal fluid of women with and without endometriosis, collected during surgery, and mRNA was extracted for analysis of Toll-like receptors expression by reverse-transcriptase polymerase chain reaction. Results Patients with endometriosis presented regulatory T cells expressing a larger number and variety of Toll-like receptors when compared to regulatory T cells from patients in the Control Group. Toll-like receptor-1 and Toll-like receptor-2 in regulatory T cells were expressed in both groups. All other expressed Toll-like receptors types were only found in regulatory T cells from the Endometriosis Group. Conclusion Patients with endometriosis had peritoneal regulatory T cells expressing various Toll-like receptors types.
RESUMO Objetivo Analisar e comparar a expressão de receptores do tipo Toll por células T reguladoras presentes no líquido peritoneal de pacientes com endometriose. Métodos Células T reguladoras foram isoladas do líquido peritoneal de mulheres com e sem endometriose, coletadas durante a cirurgia, e o RNAm foi extraído para análise da expressão de receptores do tipo Toll por reação em cadeia da polimerase com transcriptase reversa. Resultados Pacientes com endometriose apresentaram células T reguladoras expressando maior número e variedade de Toll por células quando comparadas com T reguladoras de pacientes do Grupo Controle. Receptores do tipo Toll-1 e receptores do tipo Toll-2 foram expressos em ambos os grupos. Todos os outros tipos de receptores Toll foram encontrados expressos apenas em células T reguladoras do grupo com endometriose. Conclusão Pacientes com endometriose apresentaram células T reguladoras peritoneais expressando vários tipos de receptores tipo Toll.
Assuntos
Humanos , Feminino , Adolescente , Adulto , Adulto Jovem , Líquido Ascítico/patologia , Linfócitos T Reguladores/química , Endometriose/patologia , Endométrio/patologia , Receptores Toll-Like/análise , Valores de Referência , Líquido Ascítico/imunologia , Índice de Massa Corporal , Estudos de Casos e Controles , Linfócitos T Reguladores/imunologia , Estatísticas não Paramétricas , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Endometriose/imunologia , Endométrio/imunologia , Escala Visual AnalógicaRESUMO
Endometriosis is a chronic gynecological disease, characterized by growth of endometrial tissue in ectopic sites due to alteration of peritoneal homeostasis and deregulation of apoptosis. Here we have examined whether TNFRp55 deficiency modulates the pro-inflammatory state and the reinnervation of endometriotic-like lesions in mice. Two-month-old female C57BL/6 mice, eight wild type (WT) and eight TNFRp55-/- (KO) were used in the study. Endometriotic-like lesions were induced experimentally. The right uterine horn was removed from the animal, divided longitudinally, cut in three square pieces and sutured to the intestine mesentery. After 4 weeks, the lesions and the peritoneal fluid were collected. The level of TNFα in the peritoneal fluid was evaluated by enzyme-linked immunosorbent assay (EIA). The expressions of COX2, GRα and GRß were evaluated in the lesions by western blot and immunohistochemistry. ß-III TUBULIN, BDNF and NGF protein concentrations were evaluated in the lesions by western blot. Gene expression of Pgp 9.5, SP and Th was analyzed by RT-PCR, whereas relative concentrations of TRKA, NTRp75, phosphorylated NFκB (pNFκB) and total NFκB in lesions were measured by EIA. Compared with the WT group, the KO mice showed lower TNFα levels in the peritoneal fluid and lower numbers of COX2 immunoreactive cells along with increased expression of GRα, ß-III TUBULIN, Pgp 9.5, SP, Th, BDNF, NGF, NTRp75 and pNFκB in the lesions. Future histological studies will be necessary to confirm the sensory/sympathetic imbalance in the endometriotic-like lesions of the KO mice. Our results suggest that a reduced inflammatory state promotes reinnervation of endometriotic-like lesions in TNFRp55-/- mice. Chronic deregulation of TNF receptors can have serious consequences for women with advanced endometriosis.
Assuntos
Endometriose/imunologia , Endometriose/metabolismo , Endométrio/inervação , Endométrio/metabolismo , Inflamação/imunologia , Inflamação/metabolismo , Receptores Tipo I de Fatores de Necrose Tumoral/deficiência , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Receptores Chamariz do Fator de Necrose Tumoral/deficiência , Receptores Chamariz do Fator de Necrose Tumoral/metabolismo , Animais , Apoptose/genética , Apoptose/fisiologia , Western Blotting , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Endométrio/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
OBJECTIVE: To detect Mollicutes in women with endometriosis and healthy peritoneal tissues and evaluate the participation of these bacteria in the immune response during endometriosis. DESIGN: Cross-sectional study. SETTING: University hospitals. PATIENT (S): Women with endometriosis (n = 73) and without endometriosis (n = 31). INTERVENTION(S): Endocervical swabs, peritoneal fluid, and biopsied lesions of endometriosis of women with endometriosis (study group) and healthy peritoneal tissues (control group) were collected during surgery. Clinical characteristics were registered before surgery. MAIN OUTCOME MEASURE(S): We determined the infectious agents with the use of quantitative polymerase chain reaction (PCR). The cytokine secretion profile was determined with the use of Luminex. The expression of immune response related genes was determined with the use of a PCR array kit. RESULT(S): All target microorganisms were detected at least once in the swab samples analyzed. It was possible to observe higher diversity of microorganisms in the samples of swab and peritoneal fluid in the study group compared with the control. Ureaplasma parvum was associated with the severity of the symptom dyspareunia. Mycoplasma genitalium was associated with higher production of interferon-γ and interleukin-1ß. Genes of inflammatory response activation and antigen presentation were up-regulated in biopsied tissue of women with endometriosis. In women with endometriosis, peritoneal fluid cells showed a down-regulation of genes associated with the inflammatory response. This down-regulation profile was higher in presence of M. genitalium. CONCLUSION(S): Mycoplasma genitalium may play a key role in the immune tolerance process and, especially, the aggravation of this profile. More studies are needed to understand this immune tolerance profile of bacterial infections.
Assuntos
Endometriose/imunologia , Endometriose/microbiologia , Tolerância Imunológica , Infecções por Mycoplasma/imunologia , Infecções por Mycoplasma/microbiologia , Mycoplasma genitalium/imunologia , Adolescente , Adulto , Líquido Ascítico/imunologia , Líquido Ascítico/microbiologia , Estudos de Casos e Controles , Colo do Útero/imunologia , Colo do Útero/microbiologia , Estudos Transversais , Endometriose/genética , Endometriose/metabolismo , Feminino , Regulação da Expressão Gênica , Hospitais Universitários , Interações Hospedeiro-Patógeno , Humanos , Tolerância Imunológica/genética , Mediadores da Inflamação/metabolismo , Interferon gama/genética , Interferon gama/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Pessoa de Meia-Idade , Infecções por Mycoplasma/genética , Infecções por Mycoplasma/metabolismo , Mycoplasma genitalium/patogenicidade , Adulto JovemRESUMO
BACKGROUND: Endometriosis is defined as the presence of endometrial glands and stroma at ectopic locations. Although the prevalence of endometriosis is as high as 35%-50%, its pathogenesis remains controversial. An increasing number of studies suggest that changes in immune reactivity may be primarily involved in the development of endometriosis development. In this sense, it has been strongly suggested that a fundamental part of immunologic system, the natural killer cells (NK cells), are an important part of this process. NK cells, a component of the innate immune system, have been extensively studied for their ability to defend the organism against infections and malignancy. Recent studies have shown that IL-2-activated NK (A-NK) cells are able to attack and destroy tumors in lungs and livers of mice, demonstrating the therapeutic potential of these cells. Similarly to metastatic tumor cells, endometrial cells are able to adhere, infiltrate and proliferate at ectopic locations. Therefore, in this study, we evaluated the ability of adoptively transferred and endogenous NK cells to infiltrate endometriosis lesions. METHODS: As NK cells donors were used C57BL/6 B6. PL- Thy 1.1 female mice. As uterine horns donors were used C57/BL6+GFP female mice and as endometriosis recipients C57BL/6 Thy1.2 female mice. Endometriosis induction was made by injection of endometrial tissue fragments. After 4 weeks, necessary for endometriosis lesions establishment the animals were divided in 3 experimental groups with 10 animals each. Group 1 received i.v doses of 5x106 A-NK in 200µl RPMI; Group 2 received i.p dose of 5x106 A-NK in 200µl RPMI and Group 3 received i.p dose of IL2 (0.5 mL RPMI containing 5.000U of IL2). RESULTS: Our data show that exogenous A-NK cells injected via ip combined with endogenous A-NK cells seems to be the most efficient way for activated NK cells track and infiltrate endometriosis. CONCLUSION: For the first time, it was shown that both endogenous as exogenous A-NK cells are able to track, migrate and infiltrate endometriosis lesion. This seems to be a promising result, and if confirmed the efficiency of A-NK cells in killing endometriosis lesions, maybe in the future we could use this approach as an alternative treatment for women with endometriosis.
Assuntos
Movimento Celular , Endometriose/imunologia , Endometriose/patologia , Células Matadoras Naturais/imunologia , Ativação Linfocitária/imunologia , Animais , Contagem de Células , Movimento Celular/efeitos dos fármacos , Feminino , Proteínas de Fluorescência Verde/metabolismo , Humanos , Injeções Intraperitoneais , Injeções Intravenosas , Interleucina-2/farmacologia , Células Matadoras Naturais/efeitos dos fármacos , Ativação Linfocitária/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Camundongos TransgênicosRESUMO
Endometriosis is a gynecological inflammatory and estrogen dependent pathology, defined as the presence of endometrial tissue outside the uterine cavity. The two most common symptoms of endometriosis are pelvic pain and infertility. This pain can be so intense that it affects the quality of life of women, from their relationships to their daily activities. Approximately 10% of women of reproductive age suffer from this disease. The main objective of this work was to investigate which immune system abnormalities present in patients with endometriosis that prevents ectopic endometrial tissue removal. Here we describe a series of changes in the different types of leukocytes, cytokines and factors that regulate the immune response seen in patients with endometriosis and the mechanisms by which these changes, not only favor "immunological tolerance" to the endometrial implants but at the same time stimulate the development of the disease by increasing cell proliferation and angiogenesis and inhibition of apoptosis ectopic endometrial tissue.