Glycosaminoglycan Distribution in the Rat Uterine Cervix During the Estrous Cycle

OBJECTIVE:

To analyze the amount of glycosaminoglycans in the uterine cervix during each phase of the rat estrous cycle.

DESIGN:

Based on vaginal smears, forty female, regularly cycling rats were divided into four groups (n = 10 for each group): GI – proestrous, GII – estrous, GIII – metaestrous and GIV – diestrous. Animals were sacrificed at each phase of the cycle, and the cervix was immediately removed and submitted to biochemical extraction and determination of sulfated glycosaminoglycans and hyaluronic acid. The results were analyzed by ANOVA followed by the Bonferroni post-hoc test.

RESULTS:

The uterine cervix had the highest amount of total sulfated glycosaminoglycans and dermatan sulfate during the estrous phase (8.90 ± 0.55 mg/g of cetonic extract, p<0.001; and 8.86 ± 0.57 mg/g of cetonic extract, p<0.001). In addition, there was more heparan sulfate at the cervix during the proestrous phase (0.185 ± 0.03 mg/g of cetonic extract) than during any other phase (p<0.001). There were no significant changes in the concentration of hyaluronic acid in the uterine cervix during the estrous cycle.

CONCLUSION:

Our data suggest that the amount of total sulfated glycosaminoglycans may be influenced by hormonal fluctuations related to the estrous cycle, with dermatan sulfate and heparan sulfate being the glycosaminoglycans most sensitive to hormonal change.     

Chlamydia trachomatis Infection in the Female Reproductive Tract of the Rat: Influence of Progesterone on Infectivity and Immune Response

As the most common cause of sexually transmitted disease in women, chlamydial infections can lead to pelvic inflammatory disease, infertility, and ectopic pregnancy. To better understand the role played by sex hormones in modulating the immune response of the genital tract to microbial infections, we have developed a rat model to study Chlamydia trachomatis infection. Inbred female Lewis rats were primed with progesterone and inoculated by intrauterine instillation of C. trachomatis (mouse pneumonitis strain MoPn) into each uterine horn. When infected animals were examined for the presence of chlamydial antigens 14 days postinfection, both the uterus and vagina were found to be positive compared to those of saline-treated animals, which did not show specific staining. The involvement of local and systemic immune systems following chlamydial infection was determined by analyzing major histocompatibility complex (MHC) class II expression in the reproductive tract and lymphocyte proliferation in response to mitogenic and chlamydia-specific stimulation of cells from the spleen and lymph nodes (LN) draining the reproductive tract. Enhanced proliferation was observed in LN following mitogenic but not antigenic (MOMP [major outer membrane protein]) stimulation. In contrast, spleen cell proliferation was lower in chlamydia-infected rats than in saline-treated controls. MHC class II expression, an indicator of inflammatory responses, was upregulated in the uterus, on glandular epithelial cells, and adjacent to glands in response to chlamydial infection. In other experiments, when rats were infected at estrus and diestrus without prior progesterone priming, chlamydial inclusions were not detected in either the uterus or vagina. However, enhanced lymphocyte proliferation was observed in response to mitogenic and MOMP stimulation in the reproductive tract-draining LN from estrous and diestrous animals. These findings indicate that under appropriate endocrine conditions, the rat uterus is susceptible to C. trachomatis infection and that immune responses to this pathogen can be detected locally and systemically. Further, they suggest that clearance of the infection from the reproductive tract involves immune cells from the LN draining the reproductive tract.

Chlamydia trachomatis is an obligate intracellular gram-negative bacteria that is known to infect the genital tract and ocular epithelium. Disease caused by this organism can range from acute self-limiting infection to chronic inflammatory conditions, which can result in pelvic inflammatory disease, infertility, and ectopic pregnancy (12). Chlamydial infection of the urogenital tract is currently the leading cause of sexually transmitted disease in adolescents and women in the United States and Europe (11). Despite the recognition of chlamydial infection as a major public health concern, immune responses to human chlamydial infection are not well understood. Both T-cell-mediated and humoral responses are known to be involved in resolution of chlamydial infection. However, protective immunity developed as a result of chlamydial infection is transient, and reinfections are common (12).Animal models to study chlamydial infections have been developed in the mouse, guinea pig, marmoset, and nonhuman primate (13, 15). In mice, for example, inoculations of chlamydiae via intravaginal and intrauterine routes are known to cause cervicitis or salpingitis, respectively (17, 18). In this species, intravaginal infection with human serovars of chlamydiae was dependent on pretreatment with progesterone (18). In the guinea pig, however, pretreatment with estradiol markedly enhanced the course of infection with guinea pig inclusion conjunctivitis (16). In all cases, endocrine balance at the time of exposure to chlamydiae appeared to play a crucial role for host susceptibility.Previous work from our laboratory has demonstrated that sex hormones regulate the mucosal immune response in the female reproductive tract. Using the rat as a model system, we and others have shown that stage of the estrous cycle and treatment with sex hormones, specifically estradiol and progesterone, influence both the afferent and efferent arms of the immune system (for a review see reference 19). For example, antigen presentation, immunoglobulin A (IgA) transport, immune cell traffic into the reproductive tract, and cytokine production in the uterus and vagina have been shown to be under hormonal control. The aims of the present study were (i) to determine if infection could be successfully established in rats following intrauterine exposure to chlamydiae with or without progesterone treatment and (ii) to determine if immune responses occur locally and at sites distal to the reproductive tract in response to chlamydial infection.     

肿瘤微环境中免疫细胞与肿瘤逃逸

肿瘤微环境是肿瘤细胞赖以生存和发展的内环境,肿瘤免疫逃逸机制与肿瘤微环境密切相关。文章综述了肿瘤微环境中的免疫细胞,如调节性T细胞、骨髓来源的抑制性细胞、选择性活化的巨噬细胞及功能受损的树突状细胞对肿瘤免疫逃逸的作用,以及如何逆转这种肿瘤微环境的免疫治疗策略。     

乙酰胆碱对大鼠体液免疫应答的影响

本文利用神经药理学的方法分别改变大鼠中枢和外周乙酰胆碱(Ach)的含量,以及阻断中枢M或N型的胆碱能受体,观察大鼠的初次体液免疫应答的变化,从而探讨Ach对体液免疫功能的影响及其影响机制。结果表明;(1)侧脑室注射Ach的合成阻断剂密胆碱后,体液免疫应答增强;侧脑室注射Ach酯酶的抑制剂新斯的明后,免疫应答减弱;(2)腹腔给予密胆碱或新斯的明,免疫应答都无明显改变;(3)侧脑室注射M型胆碱能受体拮抗剂阿托品,使体液免疫应答增强;注射N型胆碱能受体拮抗剂六烃季胺则抑制了免疫应答。以上结果提示:中枢神经系统内的Ach可影响体液免疫功能,可能具有免疫抑制效应;外周Ach可能不参与体液免疫功能的调节。中枢Ach的免疫调节作用是通过相应的胆碱能受体实现的。     

The Role of Sex Hormones and the Tissue Environment in Immune Protection Against HIV in the Female Reproductive Tract

Despite extensive studies of the mucosal immune system in the female reproductive tract (FRT) and its regulation by sex hormones, relatively little attention has been paid to the tissue environment in the FRT that regulates immune cell function. Consisting of secretions from epithelial cells (EC), stromal fibroblasts, and immune cells in tissues from the upper (Fallopian tubes, uterus, and endocervix) and lower (ectocervix and vagina) tracts, each tissue compartment is unique and precisely regulates immune cells to optimize conditions for successful pregnancy and protection against sexually transmitted diseases including HIV. Our goal in this review is to focus on the mucosal (tissue) environment in the upper and lower FRT. Specifically, this review will identify the contributions of EC and fibroblasts to the tissue environment and examine the impact of this environment on HIV‐target cells. Much remains to be learned about the complex interactions with the tissue environment at different sites in the FRT and the ways in which they are regulated by sex hormones and chemical contraceptives. Awareness of the involvement of the tissue environment in determining immune cell function and HIV acquisition is crucial for understanding the mechanisms that lead to HIV prevention, acquisition, and the development of new therapeutic modalities of immune protection.     

5—羟色胺对大鼠体液免疫应答的影响

本研究用神经药理学方法改变大鼠全身、中枢或外周5-羟色胺(5-HT)的含量,并使用5-HT受体的拮抗剂,观察初次体液免疫应答有无变化,从而探讨5-HT调节体液免疫功能的作用途径和作用机制。结果发现:大鼠腹腔注射5-羟色氨酸(5-HTP)后,体液免疫应答减弱。腹腔注射对氯苯丙氨酸(PCPA)后,免疫应答增强。用去甲丙味嗪(DMI)预处理,然后侧脑室注射5、7-双羟色胺(5、7-DHT);或者单独侧脑室注射5-HT,免疫应答都无显著变化,腹腔注射5-HT后,免疫应答减弱。给予赛庚啶阻断5-HT的受体后,免疫应答增强。本研究结果提示:5-HT对体液免疫应答具有抑制作用;此作用是通过外周途径实现的;其机制可能与免疫细胞上的5-HT受体有关。     

Regulation of Mucosal Immunity in the Female Reproductive Tract: The Role of Sex Hormones in Immune Protection Against Sexually Transmitted Pathogens

The immune system in the female reproductive tract (FRT) does not mount an attack against human immunodeficiency virus (HIV) or other sexually transmitted infections (STI) with a single endogenously produced microbicide or with a single arm of the immune system. Instead, the body deploys dozens of innate antimicrobials to the secretions of the FRT. Working together, these antimicrobials along with mucosal antibodies attack viral, bacterial, and fungal targets. Within the FRT, the unique challenges of protection against sexually transmitted pathogens coupled with the need to sustain the development of an allogeneic fetus, has evolved in such a way that sex hormones precisely regulate immune function to accomplish both tasks. The studies presented in this review demonstrate that estradiol (E₂) and progesterone secreted during the menstrual cycle act both directly and indirectly on epithelial cells, fibroblasts and immune cells in the reproductive tract to modify immune function in a way that is unique to specific sites throughout the FRT. As presented in this review, studies from our laboratory and others demonstrate that the innate and adaptive immune systems are under hormonal control, that protection varies with the stage of the menstrual cycle and as such, is dampened during the secretory stage of the cycle to optimize conditions for fertilization and pregnancy. In doing so, a window of STI vulnerability is created during which potential pathogens including HIV enter the reproductive tract to infect host targets.     

Mucosal Immunity in the Human Female Reproductive Tract: Cytotoxic T Lymphocyte Function in the Cervix and Vagina of Premenopausal and Postmenopausal Women

PROBLEM: To investigate the mucosal immune system in the cervix and vagina of premenopausal women in terms of immune cells present and cytolytic capacity of mucosal CD3+ T cells in the lower reproductive tract. METHODS: Fresh tissue fragments prepared by vibratome sectioning were analyzed for the presence of immune cells by confocal scanning laser microscopy (CSLM). Isolated reproductive tract cells prepared by enzymatic digestion were analyzed for CD3+ T cell phenotype by FACS analysis and for cytolytic function by an anti-CD3 mAb mediated redirected lysis assay. RESULTS: As determined by CSLM, CD3+ cells as well as macrophages and dendritic cells are distributed throughout the lower female reproductive tract in both the epithelium and subepithelial mucosa. It was found that cervical and vaginal tissues from pre- and post-menopausal women contain CD3+ T cells (CTL) that have cytolytic activity, when measured in an antigen non-specific anti-CD3 mAb mediated redirected lysis assay. CONCLUSIONS: These results indicate that the lower reproductive tract of women is immuno-competent as judged by the presence of CD3, CD4, CD8, macrophage, and dendritic cells in the endocervix, ectocervix, and vagina of premenopausal and postmenopausal women. Further, these studies demonstrate that CD3+ T cells with cytolytic activity are present in the cervix and vagina during the proliferative and secretory phases of the menstrual cycle and following menopause.     

Quiescent Tissue Stem Cells Evade Immune Surveillance

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大鼠生后发育期间胃肠道生长抑素,5-羟色胺及胃泌素免疫反应细胞的分布及形态学观察

本文用免疫组织化学 PAP法对正常 Wistar大鼠生后发育期间胃肠道生长抑素 ( SS)、5 -羟色胺( 5 - HT)及胃泌素 ( Gas)免疫反应 ( IR)细胞进行了定位研究和形态学观察。结果显示 ,SS- IR细胞于生后 1d已分布于胃肠各段。生后 1d- 4 5 d,胃窦部的 SS- IR细胞随生后龄的增长而逐渐增多 ,而其余肠道的 SS- IR细胞均以 18d为最多 ;5 - HT- IR细胞在生后发育期间可见于胃肠道各段 ,以十二指肠分布最为密集。生后发育期间的 5 - HT- IR细胞均以 18d数量最多 ;生后发育期间胃肠道的 Gas- IR细胞仅见于胃、十二指肠和空肠 ,以18d时胃窦和十二指肠的密度最大。胃肠道中 SS-、5 - HT、及 Gas- IR细胞的形态多种多样。 SS- HT- IR细胞除有突起伸至腔面外 ,尚有突起与邻与细胞接触。结果提示 ,三种 IR细胞在生后继续发生变化 ,表明各种内分泌激素在个体发育期间对机体的进一步发育起着重要作用     

Secretory Component Production by Polarized Epithelial Cells from the Human Female Reproductive Tract

At mucosal surfaces, the polymeric Ig receptor (pIgR) is responsible for transporting polymeric IgA across epithelial cells. The purpose of this study was to determine whether normal epithelial cells from the female reproductive tract form tight junctions and produce secretory component, the external domain of the pIgR. Uterine, cervical and vaginal tissues from women at different stages of the menstrual cycle and following menopause were used to prepare purified epithelial cell sheets, which were cultured in cell chambers. Transepithelial resistance was measured and the media from apical and basolateral compartments assayed for secretory component. Secretory component produced by uterine epithelial cells accumulated preferentially in apical compartment and correlated with increased transepithelial resistance. Seeding as epithelial sheets at 1×10⁶ cells/cm² of matrix coated cell chambers was required for growth. Epithelial cells from endo-cervix and ecto-cervix, but not the vagina, also showed preferential production and release of secretory component into the apical chamber. In conclusion, normal epithelial cells from the human female reproductive tract grow to confluence, become polarized and produce secretory component. Our results suggest that uterine and cervical epithelial cells play a key regulatory role in the control of IgA transcytosis from tissue into secretions.     

A Cross‐Talk of Decidual Stromal Cells,Trophoblast, and Immune Cells: A Prerequisite for the Success of Pregnancy

Embryo implantation and formation of a functional placenta are complex processes that require a plethora of regulatory mechanisms involving both mother and embryo cells. Recently, an important role in this complicated cells and factors network was assigned to the decidual stromal cells (DSC) and trophoblast cells. Decidualization includes biochemical changes that trigger DSC to produce a number of factors required for the implantation and induction of immunotolerance in maternal immune system. Immunotolerance is achieved by a cascade of strictly controlled events starting with selective homing of immune cells to the feto‐maternal site, regulated proliferation, and predominant differentiation into a regulatory type of immune cells. Furthermore, cytotoxic effector functions are reduced owing to the influence of steroid hormones, factors, cytokines, and inhibitory receptors. Altogether the entire immune system of the mother is switched to tolerogenic functional state which is a prerequisite for the successful maintenance of pregnancy.     

Secretory Immunity in the Female Reproductive Tract

PROBLEM: Antibodies and antibody-producing cells display a different and characteristic distribution in body fluids and tissues. METHOD: We have investigated the tissues of the female reproductive tract to determine whether the distribution of immunoglobulin-producing cells and the contents of cervical secretions were similar to those found in tissues of the secretory immune system. RESULTS: Immunohistochemical examinations of female genital tissues revealed the presence of plasma cells that secrete IgA (and in lower numbers IgM and IgG) especially in the subepithelial layers of the uterine endo- and ectocervix, fallopian tubes, and vagina. Both IgA1- and IgA2-producing plasma cells were found in approximately equal proportions. The presence of J-chain in the IgA-secreting cells suggests the synthesis of polymeric IgA (plgA). Epithelial cells lining the fallopian tube and endocervix were positive for secretory component (SC), which is required for the transepithelial transport of pIgA into external secretions. Cervical mucus was collected and the molecular forms of IgA were separated using column chromatography. Approximately 80% of IgA in cervical mucus was polymeric compared with 55% in the vaginal fluid. CONCLUSIONS: These data indicate that all effector components of the mucosal immune system are present in the female reproductive tract. The immunization routes that lead to a secretory IgA (S-IgA) response need to be further explored.     

Neurosecretory cells of the amygdaloid Complex during Estrous Cycle

Ultrastructure of neurosecretory cells of the dorsomedial nucleus of the cerebral amygdaloid complex (one of the main zones of sexual dimorphism) was studied in different phases of the estrous cycle. The characteristics of the light and dark cells change depending on the concentrations of sex steroids during estrus and metestrus.__________Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 139, No. 2, pp. 231–233, February, 2005     

Immune interactions at the maternal-fetal interface: a focus on antigen presentation

Problem: Viruses and fetuses face similar immunologic challenges. Each must evade immune detection and destruction. The virus must avoid host recognition of intracellular infection; the fetus allogenic recognition. Each has manipulated the process of antigen presentation to allow survival in an immunologic environment otherwise predictably hostile. How have these approaches co‐evolved? What can they teach us about viral pathogenesis and immunologic interactions at the maternal‐fetal interface? Method of study: Review of relevant literature. Results: Special classical and non‐classical MHC class I products are spared from downregulation in the placenta and from viral immunoevasive strategies. Conclusions: Viruses rely upon some of the same strategies to avoid immune detection as do trophoblast cells. In the future, viral infections may prove a useful tool for studies of immunology at the maternal‐fetal interface.     

An Examination of the Development and Localization of Key Immune Cells in Developing Pouch Young of the Red-Tailed Phascogale (Phascogale calura)

Cells expressing the surface markers CD3, CD4, CD79b, IgM, MHC class II, and ModoUG (nonclassical MHC class I) were detected in red-tailed phascogale tissues using immunohistochemistry, and the appearance and localization of cells observed here was consistent with previous observations in other marsupial species. CD3⁺ cells were first detected at one day postpartum (dpp) in the thymus, followed by ModoUG⁺ cells at 5–7 dpp in the thymus and lymph nodes. CD79b⁺ cells were first detected at 12–14 dpp in bone marrow, spleen, and lymph nodes. IgM⁺ cells were first detected at 12–14 dpp in thymus, bone marrow, spleen, and lymph nodes. MHC class II⁺ cells were first detected at 12–14 dpp in thymus, bone marrow, and lymph nodes. CD4⁺ cells were detected in adult thymus and spleen only. The presence of the mature immune cell populations and their localization to characteristic T and B cell zones in mature lymphoid tissues with normal histological structure indicates that red-tailed phascogales develop immunocompetence by the end of pouch life. Anat Rec, 302:1985–2002, 2019. © 2019 American Association for Anatomy     

ORIGINAL ARTICLE: RCAS1 Decidual Immunoreactivity during Stillbirth: Immune Cell Presence and Activity

Problem Alterations in RCAS1 (a receptor‐binding cancer antigen expressed on SiSo cells) expression in the placenta and decidua may be related to the regulation of the process of maternal immune tolerance against fetal antigens. Moreover, it has been demonstrated that the occurrence of the spontaneous beginning of stillbirth is related to a decrease in the placental expression of RCAS1. There are no data currently available on the immune processes in decidua during stillbirth. The aim of this study was to evaluate the RCAS1 immunoreactivity level in decidua and to identify the cytotoxic immune cells present during labor, induced after intrauterine fetal death either with a combination of oxytocin (OT) and prostaglandins or with OT alone; a further objective was to assess the potential impact of these molecular alterations on the effectiveness of stillbirth induction. Methods The immunoreactivity of RCAS1, CD3, CD56, CD69, and CD25 was assessed by immunohistochemistry in 31 decidual samples derived from patients in whom the stillbirth occurred before the onset of labor. Results The RCAS1 immunoreactivity level was higher in a statistically significant manner in decidual tissue samples derived from patients in whom OT alone proved insufficient to induce labor after the diagnosis of intrauterine fetal death but required additionally the use of prostaglandins when compared with samples from women in whom stillbirth was induced successfully with OT alone. However, we did not observe any differences either in CD56 and CD3 positive cell presence or in CD25 and CD69 antigen immunoreactivity in the respective decidua of these two groups of patients. Conclusion The level of RCAS1 in decidua seems to influence the effectiveness of stillbirth induction.     

渗透压对未成熟树突状细胞生物力学特性和免疫学功能的影响

目的从力学生物学角度研究渗透压对未成熟树突状细胞(immature dendritic cells,im DCs)生物力学特性和免疫功能的影响。方法不同渗透压处理im DCs后,CCK-8检测细胞存活率,激光共聚焦显微镜观察细胞骨架结构的变化,细胞电泳仪检测细胞电泳迁移率的改变,荧光偏振法检测细胞的膜流动性,实时荧光定量PCR检测免疫相关分子的表达变化,流式细胞仪检测细胞的抗原吞噬能力。结果高、低渗均会改变im DCs的F-actin结构,甚至诱导细胞凋亡。低渗组电泳率显著高于等渗组,高渗组电泳率低于等渗组(P<0.05)。荧光偏振结果显示,高、低渗均会显著降低细胞的膜流动性(P<0.05);q PCR结果发现,高、低渗会显著上调im DCs免疫表型分子CCR7、CD40、CD205、CD11a、CD11c的表达和抗原吞噬能力(P<0.05)。结论高、低渗应激会影响im DCs生物力学特性和免疫表型分子表达,研究结果对深入理解DCs的免疫调节功能具有重要意义。