Different Expressions of T-helper Cell Subsets-Related Transcription Factors and Cytokines in Infertile Women with Endometriosis

Research Article

Austin J Reprod Med Infertil. 2021; 7(1): 1057.

Different Expressions of T-helper Cell Subsets-Related Transcription Factors and Cytokines in Infertile Women with Endometriosis

Tarokh M1, Poordast T2,3, Ghaffari-Novin M1*, are M4, Tahmasebi F4, Hesampour F4 and Gharesi-Fard B3,4,5*

1Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

2Department of Obstetrics and Gynecology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

3Infertility Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

4Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

5Department of Reproductive Biology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran

*Corresponding author: Ghaffari-Novin M, Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Gharesi-Fard B, Department of Reproductive Biology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran

Received: September 17, 2021; Accepted: November 02, 2021; Published: November 09, 2021

Abstract

Purpose: It is suggested that alterations in immunologic mechanisms intervenes in the pathophysiology of endometriosis and the disease complications. There are several published reports regarding alteration in T helper cell subsets in the pathology of reproductive organs. This study was designed to find the pattern of T helper subsets in endometriosis compared with non-endometriosis women.

Methods: Expression of T helper subset-specific transcription factors (TBET, GATA3, RORC, FOXP3) and related cytokines, including IL-4, IL-10, IL-17, IL-23, TNFA, IFNG and TGFB were investigated by real time-PCR in peripheral blood, peritoneal fluid, eutopic and ectopic endometrium.

Results: The expressions of RORC, FOXP3 and all the studied cytokines were significantly increased in blood mononuclear cells of endometriosis group. The expression levels of TBET, FOXP3, IL-17A and TGFB were significantly elevated in peritoneal fluid mononuclear cells of endometriosis women. Besides, the expression levels of GATA3, RORC, FOXP3, IL-4, TNFA, and IFNG were significantly increased in eutopic endometrium of endometriosis patients. Moreover, GATA3, FOXP3 and IL-4 showed increased expressions in ectopic endometrium, while TBET, IL-23 and IFNG were significantly decreased in comparison with endometrium of non-endometriosis women. Finally, the expression levels of TBET, RORC, IL-23, TNFA, and IFNG significantly declined in ectopic endometrium in comparison with eutopic endometrium among endometriosis women.

Conclusions: Based on the present study results, it seems Th17, Th1, Th2, Treg cells and their related cytokines may play influencing role in progression of endometriosis and its symptoms including infertility. Indeed, cytokine network imbalance may cause the disease progression in patients during several years.

Keywords: Endometriosis; Infertility; T cell subsets; Transcriptional Factors

Abbreviations

PB: Peripheral Blood; PF: Peritoneal Fluid; Th: T helper; Treg: T regulatory; IL: Interleukine; IFN: Interferone; TGF: Transforming Growth Factor; TF: Transcriptional Factor; TNF: Tumor Necrotizing Factor; PBMC: Peripheral Blood Mononuclear Cell; PFMC: Peritoneal Fluid Mononuclear Cell; Eeu: eutopic Endometrium; Eec: ectopic Endometrium; FSH: Follicle Stimulating Hormone; LH: Luteinizing Hormone; PRL: Prolactin; TSH: Thyroid Stimulating Hormone

Introduction

Endometriosis is a common chronic inflammatory pelvic disease in women at reproductive age with two main manifestations, including pain and infertility [1,2]. This disease is defined as the presence of endometrial tissue out of the uterine cavity [2]. Although the exact etiology of endometriosis is not yet well known, the most accepted theory is retrograde menstruation, which could initiate immune system responses to ectopically implanted endometrial cells on the peritoneal lining among 10 to 15% of women [3,4]. It is suggested that alterations in immunologic mechanisms intervenes in the pathophysiology of endometriosis and the disease complications [3]. In women with endometriosis, significant changes have been reported in population of immune cell subsets and types of responses in the Peripheral Blood (PB), Peritoneal Fluid (PF) as well as endometrial tissues [5,6]. Various leukocytes reside in eutopic endometrium and also endometriotic implants, and produce different cytokines and other immune mediators [5,7]. There are several reports regarding deviation of Th cells responses in endometriosis [8- 10]. While predominance of Th1 cells may promote inflammation, increase in Th2 cells modulates inflammation in affected areas. Th1 cells potentiate inflammation by producing IFN-γ. Depending on the level of secretion, IFN-γ and TNF-a are proposed to have dual effects on early stage of gestation and implantation [11]. On the other hand TNF-a has toxicity on sperm and egg cells and activates Th1cells [12]. TNF-a may also be produced by Th1. Th2 cells secrete IL-4, IL-10, and TGF-β as anti-inflammatory cytokines. Increase of IL-4 and IL- 10 is reported in some research on endometriosis [10]. In addition, Th17 and regulatory T (Treg) cell subsets have been also proposed to play roles in pathogenesis of endometriosis [13-15]. Th17 cells may trigger inflammation by production of neutrophil attractive chemokines [16]. It is argued that Th17 cells deal with induction of many autoimmune diseases, including arthritis, inflammatory bowel disease, psoriasis, uveitis, and multiple sclerosis [17,18]. IL-17A is the most important cytokine produced by Th17 cells, which mediates neutrophilic inflammation [19].

Increase in Treg cells inhibits the recruitment of effector immune cells. Treg cells also produce anti-inflammatory IL-10 and TGF-β cytokines [4]. It is reported that women with endometriosis show decreased level of cytokines IL-17 and TGF-β some weeks after laparoscopy [20]. This is also followed by increase in pregnancy rate during next months.

Moreover, it is well documented that Th cells and their related cytokines cooperate with each other as a network. So, disturbance in Th subsets could be accounted as a major immunological etiology for many infertility-related diseases including endometriosis [21,22]. In line with this idea, several studies have shown that the proportion of immune cells and cytokine production in women with endometriosis has altered in comparison with healthy women [23,24]. In addition to signature cytokines, investigation of Th subset-specific Transcription Factors (TFs) has been also conducted with the aim of evaluating the Th balance in endometriosis [25,26].

Interestingly, there is no published data regarding the simultaneous investigation and comparison of the Th subsets balance in all the potential samples, including PB, PF, intra (eutopic), and extra (ectopic) uterine endometrial tissue samples in endometriosis patients. In the previous published research, only one or two source of PB, PF, eutopic or ectopic uterine endometrial tissues have been investigated. So, in the present study, the expression of TFs and cytokines related to Th1, Th2, Th17, and Treg in PB, PF, and endometrial tissues were simultaneously investigated to elucidate the pattern and the role of T cell subsets involved in endometriosis pathogenesis. For this purpose, we analyzed and compared genes expression of TBET, GATA3, RORC, FOXP3, IL-4, IL-10, IL-17, IL- 23, TNFA, IFNG, and TGFB from endometriosis and healthy women in all the mentioned samples.

Materials and Methods

Subjects

This study was approved by the Medical Ethics Committees of both Shahid Beheshti University of Medical Sciences, Tehran, Iran (IR.SBMU.SM.REC.1394.79) and Shiraz University of Medical Sciences, Shiraz, Iran (IR.SUMS.REC.1395.S307). All participants read and signed written informed consent before entering this research study and sampling. Clinical and demographic features of the participants are shown in Table 1.