RESUMEN
Regulation of endometrial (EM) CD8+T cells is essential for successful reproduction and protection against pathogens. Suppression of CD8+T cells is necessary for a tolerogenic environment that promotes implantation and pregnancy. However, the mechanisms regulating this process remain unclear. Sex hormones are known to control immune responses directly on immune cells and indirectly through the tissue environment. When the actions of estradiol (E2), progesterone (P) and TGFß on EM CD8+T cells were evaluated, cytotoxic activity, perforin and granzymes were directly suppressed by E2 and TGFß but not P. Moreover, incubation of polarized EM epithelial cells with P, but not E2, increased TGFß secretion. These findings suggest that E2 acts directly on CD8+T cell to suppress cytotoxic activity while P acts indirectly through induction of TGFß production. Understanding the mechanisms involved in regulating endometrial CD8+T cells is essential for optimizing reproductive success and developing protective strategies against genital infections and gynecological cancers.
Asunto(s)
Endometrio/citología , Endometrio/inmunología , Estradiol/metabolismo , Progesterona/metabolismo , Linfocitos T Citotóxicos/inmunología , Factor de Crecimiento Transformador beta1/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Citotoxicidad Inmunológica/inmunología , Implantación del Embrión/inmunología , Implantación del Embrión/fisiología , Femenino , Granzimas/biosíntesis , Humanos , Persona de Mediana Edad , Perforina/biosíntesis , EmbarazoRESUMEN
A health care crisis such as the coronavirus disease 2019 (COVID-19) pandemic requires allocation of hospital staff and resources on short notice. Thus, new and sometimes less experienced team members might join the team to fill in the gaps. This scenario can be particularly challenging in endovascular stroke treatment, which is a highly specialized task that requires seamless cooperation of numerous health care workers across various specialties and professions. This document is intended for stroke teams who face the challenge of integrating new team members into endovascular stroke-treatment workflows during the COVID-19 pandemic or any other global health care emergency. It discusses the key strategies for smooth integration of new stroke-team members in a crisis situation: 1) transfer of key knowledge (simple take-home messages), 2) open communication and a nonjudgmental atmosphere, 3) strategic task assignment, and 4) graded learning and responsibility. While these 4 key principles should generally be followed in endovascular stroke treatment, they become even more important during health care emergencies such as the COVID-19 pandemic, when health care professionals have to take on new and additional roles and responsibilities in challenging working environments for which they were not specifically trained.
Asunto(s)
SARS-CoV-2 , Accidente Cerebrovascular/terapia , COVID-19 , Humanos , Flujo de TrabajoRESUMEN
Dendritic cells (DCs) throughout the female reproductive tract (FRT) were examined for phenotype, HIV capture ability and innate anti-HIV responses. Two main CD11c+ DC subsets were identified: CD11b+ and CD11blow DCs. CD11b+CD14+ DCs were the most abundant throughout the tract. A majority of CD11c+CD14+ cells corresponded to CD1c+ myeloid DCs, whereas the rest lacked CD1c and CD163 expression (macrophage marker) and may represent monocyte-derived cells. In addition, we identified CD103+ DCs, located exclusively in the endometrium, whereas DC-SIGN+ DCs were broadly distributed throughout the FRT. Following exposure to GFP-labeled HIV particles, CD14+ DC-SIGN+ as well as CD14+ DC-SIGN- cells captured virus, with â¼30% of these cells representing CD1c+ myeloid DCs. CD103+ DCs lacked HIV capture ability. Exposure of FRT DCs to HIV induced secretion of CCL2, CCR5 ligands, interleukin (IL)-8, elafin, and secretory leukocyte peptidase inhibitor (SLPI) within 3 h of exposure, whereas classical pro-inflammatory molecules did not change and interferon-α2 and IL-10 were undetectable. Furthermore, elafin and SLPI upregulation, but not CCL5, were suppressed by estradiol pre-treatment. Our results suggest that specific DC subsets in the FRT have the potential for capture and dissemination of HIV, exert antiviral responses and likely contribute to the recruitment of HIV-target cells through the secretion of innate immune molecules.
Asunto(s)
Células Dendríticas/inmunología , Genitales Femeninos/inmunología , Infecciones por VIH/inmunología , VIH/inmunología , Inmunidad Innata , Antígeno CD11c/metabolismo , Moléculas de Adhesión Celular/metabolismo , Células Cultivadas , Quimiocina CCL2/metabolismo , Células Dendríticas/virología , Elafina/metabolismo , Estradiol/farmacología , Femenino , VIH/patogenicidad , Infecciones por VIH/transmisión , Humanos , Interleucina-8/metabolismo , Lectinas Tipo C/metabolismo , Receptores de Lipopolisacáridos/metabolismo , Fagocitosis , Receptores CCR5/metabolismo , Receptores de Superficie Celular/metabolismo , Inhibidor Secretorio de Peptidasas Leucocitarias/metabolismoRESUMEN
Prevention of sexual acquisition of HIV in women requires a substantial increase in our knowledge about HIV-target cell availability and regulation in the female reproductive tract (FRT). In this study, we analyzed the phenotype and susceptibility to HIV infection of CD4(+) T cell in the endometrium (EM), endocervix (END), and ectocervix (ECT) of the FRT. We found that T helper type 17 (Th17) cells represent a major subset in FRT tissues analyzed and that Th17 cells were the main CD4(+) T-cell population expressing C-C motif chemokine receptor 5 (CCR5) and CD90. In premenopausal women, CD4(+) T cells and Th17 cells, in particular, were significantly lower in EM relative to END and ECT. Th17 cells were elevated in EM from postmenopausal women relative to premenopausal tissues but not changed in END and ECT. Susceptibility of CD4(+) T cells to HIV infection measured as intracellular p24 was lowest in the EM and highest in the ECT. Additionally, we found that Th17 cells co-expressing CCR5 and CD90 were the most susceptible to HIV infection. Our results provide valuable information for designing preventive strategies directed at targeting highly susceptible target cells in the FRT.
Asunto(s)
Cuello del Útero/inmunología , Endometrio/inmunología , Infecciones por VIH/inmunología , Posmenopausia/inmunología , Células Th17/inmunología , Adulto , Anciano , Cuello del Útero/patología , Susceptibilidad a Enfermedades/inmunología , Susceptibilidad a Enfermedades/patología , Endometrio/patología , Femenino , Regulación de la Expresión Génica/inmunología , Infecciones por VIH/patología , Humanos , Persona de Mediana Edad , Receptores CCR5/inmunología , Células Th17/patología , Antígenos Thy-1/inmunologíaRESUMEN
The complexity of the human female reproductive tract (FRT) with its multiple levels of hormonally controlled immune protection has only begun to be understood. Dissecting the functions and roles of the immune system in the FRT is complicated by the differential hormonal regulation of its distinct anatomical structures that vary throughout the menstrual cycle. Although many fundamental mechanisms of steroid regulation of reproductive tract immune function have been determined, the effects of exogenous synthetic steroids or endocrine disruptors on immune function and disease susceptibility in the FRT have yet to be evaluated in detail. There is increasing evidence that environmental or synthetic molecules can alter normal immune function. This review provides an overview of the innate and adaptive immune systems, the current status of immune function in the FRT and the potential risks of environmental or pharmacological molecules that may perturb this system.
Asunto(s)
Disruptores Endocrinos/farmacología , Genitales Femeninos/inmunología , Inmunidad Mucosa/efectos de los fármacos , Membrana Mucosa/inmunología , Animales , Exposición a Riesgos Ambientales , Femenino , Genitales Femeninos/citología , Genitales Femeninos/efectos de los fármacos , Genitales Femeninos/metabolismo , Humanos , Sistema Inmunológico/citología , Sistema Inmunológico/efectos de los fármacos , Sistema Inmunológico/crecimiento & desarrollo , Membrana Mucosa/efectos de los fármacos , Membrana Mucosa/metabolismo , Reproducción/efectos de los fármacosRESUMEN
This review examines the multiple levels of pre-existing immunity in the upper and lower female reproductive tract. In addition, we highlight the need for further research of innate and adaptive immune protection of mucosal surfaces in the female reproductive tract. Innate mechanisms include the mucus lining, a tight epithelial barrier and the secretion of antimicrobial peptides and cytokines by epithelial and innate immune cells. Stimulation of the innate immune system also serves to bridge the adaptive arm resulting in the generation of pathogen-specific humoral and cell-mediated immunity. Less understood are the multiple components that act in a coordinated way to provide a network of ongoing protection. Innate and adaptive immunity in the human female reproductive tract are influenced by the stage of menstrual cycle and are directly regulated by the sex steroid hormones, progesterone and estradiol. Furthermore, the effect of hormones on immunity is mediated both directly on immune and epithelial cells and indirectly by stimulating growth factor secretion from stromal cells. The goal of this review is to focus on the diverse aspects of the innate and adaptive immune systems that contribute to a unique network of protection throughout the female reproductive tract.
Asunto(s)
Genitales Femeninos/inmunología , Infecciones/inmunología , Membrana Mucosa/inmunología , Enfermedades de Transmisión Sexual/inmunología , Inmunidad Adaptativa , Animales , Femenino , Humanos , Inmunidad Innata , Infecciones/transmisión , Embarazo , Enfermedades de Transmisión Sexual/transmisiónRESUMEN
Female reproductive tract (FRT) epithelial cells protect against potential pathogens and sexually transmitted infections. The purpose of this study was to determine if epithelial cells from the upper FRT secrete antimicrobials that inhibit reproductive tract pathogens that threaten women's health. Apical secretions from primary cultures of Fallopian tube, uterine, cervical, and ectocervical epithelial cells were incubated with Neisseria gonorrhoeae, Candida albicans (yeast and hyphal forms), human immunodeficiency virus 1 (HIV-1), and Lactobacillus crispatus before being tested for their ability to grow and/or infect target cells. Epithelial cell secretions from the upper FRT inhibit N. gonorrhoeae and both forms of Candida, as well as reduce HIV-1 (R5) infection of target cells. In contrast, none had an inhibitory effect on L. crispatus. An analysis of cytokines and chemokines in uterine secretions revealed several molecules that could account for pathogen inhibition. These findings provide definitive evidence for the critical role of epithelial cells in protecting the FRT from infections, without comprising the beneficial presence of L. crispatus, which is part of the normal vaginal microflora of humans.
Asunto(s)
Antiinfecciosos/metabolismo , Secreciones Corporales/metabolismo , Candida albicans/inmunología , Epitelio/metabolismo , VIH-1/inmunología , Lactobacillus/inmunología , Neisseria gonorrhoeae/inmunología , Enfermedades de Transmisión Sexual/inmunología , Antiinfecciosos/inmunología , Secreciones Corporales/inmunología , Candida albicans/efectos de los fármacos , Candida albicans/crecimiento & desarrollo , Técnicas de Cultivo de Célula , Procesos de Crecimiento Celular/efectos de los fármacos , Células Cultivadas , Citocinas/metabolismo , Epitelio/inmunología , Epitelio/microbiología , Epitelio/patología , Epitelio/virología , Femenino , Genitales Femeninos/patología , VIH-1/crecimiento & desarrollo , Humanos , Lactobacillus/efectos de los fármacos , Lactobacillus/crecimiento & desarrollo , Neisseria gonorrhoeae/efectos de los fármacos , Neisseria gonorrhoeae/crecimiento & desarrollo , Enfermedades de Transmisión Sexual/prevención & controlRESUMEN
Human NK cells from the decidua basalis of gravid uteri and from the cycling endometrium of women undergoing hysterectomy were isolated and compared by gene expression profiling using Affymetrix microarrays with probes representing approximately 47,400 transcripts. Substantial differences indicate that these two types of NK cells represent distinct subsets.
Asunto(s)
Decidua/inmunología , Endometrio/inmunología , Células Asesinas Naturales/metabolismo , Subgrupos Linfocitarios/metabolismo , Útero/inmunología , Separación Celular , Femenino , Perfilación de la Expresión Génica , Humanos , Células Asesinas Naturales/inmunología , Embarazo/inmunologíaRESUMEN
The goal of this study was to examine the role of E(2) in regulating innate immune protection by human uterine epithelial cells (UECs). Recognizing that UECs produce cytokines and chemokines to recruit and activate immune cells as well as viral and bacterial antimicrobials, we sought to examine the effect of E(2) on constitutive and Toll-like receptor (TLR) agonist (lipopolysaccharide (LPS) and poly (I:C))-induced immune responses. The secretion by polarized UECs in culture of interleukin (IL)-6, macrophage inhibitory factor (MIF), and secretory leukocyte protease inhibitor (SLPI) was examined as well as the mRNA expression of human beta-defensin-2 (HBD2), tumor necrosis factor (TNF)-alpha, IL-8, and nuclear factor (NF)-kB. When incubated with E(2) for 24-48 h, we found that E(2) stimulated UEC secretion of SLPI (fourfold) and mRNA expression of HBD2 (fivefold). Moreover, when antibacterial activity in UEC secretions was measured using Staphylococcus aureus, E(2) increased the secretion of soluble factor(s) with antibacterial activity. In contrast, E(2) had no effect on constitutive secretion of proinflammatory cytokines and chemokines by UECs but completely inhibited LPS- and poly (I:C)-induced secretion of MIF, IL-6, and IL-8. Estradiol also reversed the stimulatory effects of IL-1beta on mRNA expression of TNF-alpha, IL-8, and NF-kB by 85, 95, and 70%, respectively. As SLPI is known to inhibit NF-kB expression, these findings suggest that E(2) inhibition of proinflammatory cytokines may be mediated through SLPI regulation of NF-kB. Overall, these findings indicate that the production of cytokines, chemokines, and antimicrobials by UECs are differentially regulated by E(2). Further, it suggests that with E(2) regulation, epithelial cells that line the uterine cavity have evolved immunologically to be sensitive to viral and bacterial infections as well as the constraints of procreation.
Asunto(s)
Polaridad Celular/fisiología , Citocinas/inmunología , Células Epiteliales/metabolismo , Estradiol/farmacología , Inmunidad Innata , Útero/citología , Células Cultivadas , Citocinas/metabolismo , Células Epiteliales/citología , Células Epiteliales/inmunología , Estradiol/fisiología , Femenino , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Oxidorreductasas Intramoleculares/metabolismo , Lipopolisacáridos/farmacología , Factores Inhibidores de la Migración de Macrófagos/metabolismo , FN-kappa B/metabolismo , Inhibidor Secretorio de Peptidasas Leucocitarias/metabolismo , Staphylococcus aureus/fisiología , Receptores Toll-Like/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , beta-Defensinas/metabolismoRESUMEN
Neutrophils enter tissues including the uterus and are found in the endometrium in increased numbers prior to menses. In this environment, they are exposed to transforming growth factor (TGF)-beta1 produced by endometrial stromal and epithelial cells. We observed that incubation of neutrophils in vitro with TGF-beta1 at 1 pg/ml significantly reduced their secretion of lactoferrin in response to lipopolysaccharide (LPS). This effect was achieved with as little as 15 min of pretreatment with TGF-beta1. Inhibition of lactoferrin release by TGF-beta1 was observed irrespective of whether neutrophils were stimulated by ligands for Toll-like receptor (TLR)-2, TLR-4 or FPR, the G protein-coupled receptor for formylated peptides. Inhibition by TGF-beta1 was negated by SB-431542, a small molecule inhibitor that specifically blocks the kinase activity of the type I TGF-beta receptor (ALK5) In contrast to lactoferrin release, another important neutrophil function, interleukin (IL)-8 driven chemotaxis, was not affected by TGF-beta1 at 1 pg/ml or 100 pg/ml. We conclude that in tissues of the female reproductive tract, TGF-beta1 inhibition of neutrophil degranulation may prevent these cells from initiating an inflammatory response or releasing degradative enzymes that could potentially damage the oocyte or fetus.
Asunto(s)
Degranulación de la Célula/inmunología , Neutrófilos/fisiología , Factor de Crecimiento Transformador beta1/inmunología , Benzamidas/farmacología , Células Cultivadas , Quimiotaxis de Leucocito/inmunología , Dioxoles/farmacología , Relación Dosis-Respuesta Inmunológica , Femenino , Humanos , Lactoferrina/metabolismo , Lipopolisacáridos/inmunología , Neutrófilos/inmunología , Receptores de Factores de Crecimiento Transformadores beta/antagonistas & inhibidores , Proteínas Recombinantes/inmunologíaRESUMEN
BACKGROUND: Pro-inflammatory chemokines that attract and cytokines that activate immune cells contribute to normal physiological homeostasis in the female reproductive tract, and are needed to deal effectively with potential pathogenic microbes. Mucosal epithelial cells are capable of producing these factors that communicate with cells of the innate and adaptive immune systems. METHODS: Epithelial cells from Fallopian tube, endometrium and endocervix were isolated and grown to high transepithelial resistance in cell inserts from seven patients who had hysterectomies. Interleukin (IL)-8, IL-6, granulocyte colony-stimulating factor (G-CSF), monocyte chemoattractant protein-1 (MCP-1), granulocyte-macrophage colony-stimulating factor (GM-CSF), tumour necrosis factor-alpha (TNF-alpha) and macrophage inflammatory peptide-1beta (MIP-1beta) were assessed by Luminex bead analysis or enzyme-linked immunosorbent assay (ELISA) in epithelial cell conditioned media from the apical and basolateral compartments. RESULTS: With the exception of MCP-1, the seven chemokines/cytokines constitutively produced by the polarized epithelial cells were preferentially secreted apically. A concentration pattern was found in all cases, with IL-8 and IL-6 produced in the greatest quantity. CONCLUSIONS: The concentrations of IL-8, IL-6, G-CSF and MCP-1 are similar to the levels found in reproductive tract fluids of patients with infection. The constitutive secretion and compartmentalization of large quantities of bioactive chemokines and cytokines provide additional evidence for the role of epithelial cells as gatekeepers of innate immune protection in the female reproductive tract.
Asunto(s)
Quimiocinas/metabolismo , Citocinas/metabolismo , Células Epiteliales/metabolismo , Útero/citología , Polaridad Celular , Células Cultivadas , Quimiocina CCL2/metabolismo , Células Epiteliales/fisiología , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Humanos , Infecciones/patología , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Membrana Mucosa/citología , Útero/metabolismoRESUMEN
It has been demonstrated previously that mixed cell suspensions from the female reproductive tract consisting of human epithelial and stromal cells were capable of presenting foreign antigen to autologous T cells. There have been, however, no reported studies examining antigen presentation by isolated epithelial cells from the human female reproductive tract. It is now shown that freshly isolated epithelial cells from the uterine endometrium constitutively express MHC class II antigen and that class II was upregulated on cultured epithelium by interferon gamma (IFNgamma). Using a highly purified preparation, it was demonstrated that these epithelial cells were able to process and present tetanus toxoid recall antigen driving autologous T cell proliferation. Cells isolated from the basolateral sub-epithelium stroma were also potent antigen presenting cells in this model system. Thus, isolated endometrial epithelial cells were able to directly process and present antigen to T cells and may be responsible for the transcytosis and delivery of antigen to professional antigen presenting cells found in the sub-epithelial stroma.
Asunto(s)
Endometrio/inmunología , Regulación de la Expresión Génica/efectos de los fármacos , Genes MHC Clase II , Antígenos de Histocompatibilidad Clase II/biosíntesis , Interferón gamma/farmacología , Adulto , Células Cultivadas , Femenino , Humanos , Células del Estroma/inmunologíaRESUMEN
Lymphoid aggregates (LA) develop during the proliferative phase of the menstrual cycle in the human uterine endometrium (EM). They contain mostly CD8+ T cells and B cells. As these LA are absent immediately following menses, they may arise by division of cells resident in the EM, or by division of a limited number of precursor cells that traffic into the EM during the early proliferative phase of the menstrual cycle. Alternatively, they may arise by the continuous trafficking of cells into the EM throughout the proliferative phase of the menstrual cycle. In this study we investigated the distribution and frequency of CD8+ T cells in the aggregates using expression of Vbeta2 or Vbeta8 as markers of clonality and Ki-67 as a marker of dividing cells. Confocal microscopic analysis of endometrial tissues showed the random distribution of CD8+ T cells within aggregates within the same sample and in aggregates from different samples. Furthermore, comparisons of the distribution of Vbeta2 and Vb8 with expected values predicted from Poisson distribution values were not significantly different, suggesting that CD8+ T cells do not arise by division from single precursors. A low level of T-cell division within LAs was confirmed by positive staining for Ki-67. Dividing T cells were randomly dispersed throughout the LA and the frequency of dividing cells did not vary greatly between aggregates within the same tissue. Nearest-neighbour analysis of dividing cells showed no statistically significant deviations from a random distribution. Taken together, these results suggest that LA develop during the menstrual cycle largely by the trafficking of cells to nucleation sites within the EM, rather than by division of a limited number of precursor cells.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Endometrio/inmunología , Tejido Linfoide/inmunología , Ciclo Menstrual/inmunología , Subgrupos de Linfocitos T/inmunología , Apoptosis/inmunología , Agregación Celular/inmunología , División Celular/inmunología , Movimiento Celular/inmunología , Femenino , Técnica del Anticuerpo Fluorescente , Humanos , Inmunofenotipificación , Antígenos Comunes de Leucocito/análisis , Receptores de Antígenos de Linfocitos T alfa-beta/análisisRESUMEN
CD8(+) T cell phenotype and function were assessed in the female reproductive tracts (FRTs) of 3 human immunodeficiency virus (HIV)-positive patients who had undergone hysterectomy. FRT cytotoxic T lymphocyte (CTL) lytic activity from 1 patient (patient 872) was detected by using CD3-dependent redirected-lysis assay and HIV-specific assay, concomitant with the presence of CD8(+) cells. In contrast, samples from the 2 other HIV-positive patients (patients 1356 and 1364), who also were asymptomatic for HIV-associated illnesses, demonstrated no CTL activity in any solid tissue tested by either assay, despite activity by autologous peripheral blood mononuclear cells (PBMC). This absence of CTL activity was correlated with a relative absence of CD8(+) cells in the FRT, whereas CD8(+) cells were present in PBMC. Thus, CTL activity in PBMC may fail to correlate with mucosal activity. The finding of CTL activity in the FRT of patient 872 represents the first description of CTL in upper and lower FRT tissues of an HIV-positive woman.
Asunto(s)
Complejo CD3/inmunología , Genitales Femeninos/inmunología , Antígenos VIH/inmunología , Seropositividad para VIH/inmunología , Inmunidad Mucosa , Linfocitos T Citotóxicos/inmunología , Adulto , Células Cultivadas , Pruebas Inmunológicas de Citotoxicidad , Femenino , VIH/inmunología , Humanos , Histerectomía , Leucocitos Mononucleares/inmunología , FenotipoRESUMEN
PROBLEM: The objective of this study was to examine the susceptibility of rat uterine epithelial cells (UEC) to infection with Chlamydia trachomatis and to study the epithelial-stromal interactions following infection. METHOD OF STUDY: UEC were isolated from adult rats and grown in culture. Polarized, confluent monolayers of UEC were infected with 10(6) IFU/well C. trachomatis (MoPn). In order to confirm infection, MoPn was labeled with a fluorescent tracking dye, PKH-26, and then used in epithelial cell infections. Transepithelial resistances were measured prior to and following infection to test the effect of Chlamydia on the integrity of the epithelial monolayers. In other experiments, polarized epithelial cultures were infected in the presence and absence of stromal cells. Media was collected from the apical and basolateral compartments of the cultures before and after infection and analyzed for cytokines IL-1alpha and TNF-alpha. RESULTS: Epithelial cell cultures infected with PKH-26 labeled MoPn were examined 4-5 days later. Bacterial inclusions were detected inside epithelial cells indicating infection had occurred. Co-localization of PKH-26 labeled bacteria with FITC-labelled anti-Chlamydia antibody on the epithelial cells confirmed infection. No changes were found in resistance across the monolayers of epithelial cells in the presence or absence of infection. ELISA results indicate that UEC secrete IL-1alpha constitutively in citro. Stromal cells secrete very little IL-1alpha. When stromal cells were co-incubated with epithelial cells there was a decrease in the amount of IL-1alpha secreted by epithelial cells 48 hr post-infection. On the other hand, maximum TNF-alpha was found in stromal cells. both with and without infection. Epithelial cells, in these studies made very little TNF-alpha. CONCLUSIONS: These results show that primary rat epithelial cells can be infected with Chlamydia in vitro. Epithelial and stromal cells from uteri of adult rats make IL-1alpha and TNF-alpha in vitro both prior to and following infection with Chlamydia. This system can be used to analyze the role played by epithelial-stromal interactions in providing protection on this mucosal surface.
Asunto(s)
Comunicación Celular , Chlamydia trachomatis/fisiología , Interleucina-1/biosíntesis , Factor de Necrosis Tumoral alfa/biosíntesis , Útero/microbiología , Animales , Anticuerpos Antibacterianos/inmunología , Polaridad Celular , Células Epiteliales/microbiología , Femenino , Conejos , Ratas , Ratas Endogámicas LewRESUMEN
PROBLEM: The isolation of human female reproductive tract (RT) cells that maintain viability and are representative of the entire population is essential for a thorough evaluation of mucosal immunity in the reproductive tract mucosa. Here, we describe the isolation of RT cells in high yields and with high viability from the Fallopian tube, uterine endometrium, endocervix, ectocervix and vagina. METHOD OF STUDY: This cell dispersion method uses an enzyme cocktail composed of pancreatin, hyaluronidase, and collagenase (PHC), and employs a 250-microm mesh screen to facilitate cell dispersion. RESULTS: The yields of cells isolated per gram of tissue in the presence of this PHC cocktail were compared and found to be strikingly higher relative to the yields obtained with other enzyme cocktails or in the absence of enzymes. Flow cytometry was used to characterize leukocyte subsets isolated from uterine endometrium in the presence of the various enzyme cocktails. The common leukocyte antigen marker CD45, pan T-cell marker CD3, monocyte/macrophage marker CD14 and B-cell marker CD19 were retained after exposure to the PHC cocktail of enzymes. The expression of CD8 and CD4 was lost after exposure to added enzymes but regained after culture overnight. CONCLUSION: These studies demonstrate the feasibility of using enzymatic digestion for the isolation of whole populations of Fallopian tube, endometrial, cervical and vaginal cells, including leukocyte subsets in high yields, and provide a foundation for investigating mucosal immune cell function in the human female RT.
Asunto(s)
Separación Celular , Genitales Femeninos/citología , Genitales Femeninos/inmunología , Leucocitos/inmunología , Antígenos CD4/análisis , Antígenos CD8/análisis , Femenino , Humanos , Antígenos Comunes de Leucocito/análisisRESUMEN
The objective of the present study was to define the afferent arm of the mucosal immune system in the lower female reproductive tract. We report here that antigen presentation by vaginal cells is under hormonal control. When vaginal cells from ovariectomized rats treated with estradiol (0.01-10 microg) were incubated with ovalbumin-specific T cells and ovalbumin, a dose-dependent inhibition of antigen presentation was measured. In time course studies, estradiol given to ovariectomized rats inhibited vaginal cell antigen presentation within 24 h after a single injection, relative to that seen in saline controls. To determine whether changes in antigen presentation were attributable to the effect of estradiol on the number of antigen-presenting cells (APCs) in the vagina, tissues were analyzed by immunohistochemistry. Our findings indicate that estradiol inhibited antigen presentation without affecting the number of major histocompatibility complex class II positive cells and at a time when macrophage/dendritic cells/granulocytes in the vagina increase in response to estradiol treatment. Antibody neutralization studies indicated that antigen presentation by vaginal cells from ovariectomized rats is mediated through class II and involves the expression of transmembrane proteins B7.1 and B7.2. In other studies, vaginal APCs interact with thymus APCs to synergistically enhance antigen presentation under conditions in which vaginal antigen presentation is inhibited by estradiol. Analysis of conditioned media indicates that enhancement of thymus antigen presentation involves the release of a soluble factor(s) into the culture media of vaginal cells. When spleen cells were cocultured with vaginal cells from saline-treated rats, proliferation increased in the presence of concanavalin A and/or phytohemagglutinin and decreased with lipopolysaccharide, relative to spleen cells and mitogen alone. In contrast, when incubated with vaginal cells from estradiol-treated rats, spleen cell proliferation was not affected with concanavalin but was inhibited with phytohemagglutinin and lipopolysaccharide. These studies demonstrate that estradiol regulates antigen presentation by vaginal cells and that vaginal cells, in turn, influence antigen presentation, as well as B and T cell proliferation.
Asunto(s)
Presentación de Antígeno/efectos de los fármacos , Células Presentadoras de Antígenos/inmunología , Estradiol/farmacología , Vagina/citología , Vagina/inmunología , Animales , Medios de Cultivo Condicionados , Femenino , Mitosis , Membrana Mucosa/inmunología , Ovalbúmina/inmunología , Ovariectomía , Ratas , Ratas Endogámicas Lew , Bazo/citología , Bazo/inmunología , Linfocitos T/inmunología , Timo/citología , Timo/inmunologíaRESUMEN
We have used a previously described rodent model to examine the influence of hormonal environment on susceptibility and immune responses to genital Chlamydia infection. Ovariectomized rats were administered estradiol, progesterone, or a combination of both, infected with Chlamydia trachomatis via the intrauterine route, and sacrificed 5 days later. Histopathological examination showed severe inflammation in the uteri and vaginae of progesterone-treated animals, whereas animals receiving estradiol or a combination of both hormones showed no inflammation. Large numbers of chlamydiae were found in vaginal secretions of progesterone-treated and combination-treated animals, while estradiol-treated animals had none. Tissue localization showed that numerous chlamydial inclusions were present in the uterine epithelium of the progesterone group and the cervicovaginal epithelium of the combination group. Examination of the acute immune responses of the infected animals showed that maximum activation was present in the draining lymph node cells from the progesterone-treated group, and these cells were producing large amounts of interleukin-10 and gamma interferon compared to other hormone-treated groups. In contrast, spleen cell proliferation was suppressed in progesterone-treated animals compared to other hormone-treated groups. We conclude that progesterone increases and estradiol decreases susceptibility to intrauterine chlamydial infection in this rat model. Our data demonstrate that hormone environment, at the time of infection, has a profound effect on the outcome of microbial infection in the female reproductive tract.
Asunto(s)
Infecciones por Chlamydia/etiología , Infecciones por Chlamydia/inmunología , Chlamydia trachomatis , Estradiol/farmacología , Progesterona/farmacología , Animales , Cuello del Útero/inmunología , Cuello del Útero/microbiología , Cuello del Útero/patología , Infecciones por Chlamydia/patología , Chlamydia trachomatis/aislamiento & purificación , Citocinas/metabolismo , Estradiol/metabolismo , Femenino , Cuerpos de Inclusión/microbiología , Inflamación/patología , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/inmunología , Activación de Linfocitos/efectos de los fármacos , Ovariectomía , Progesterona/metabolismo , Ratas , Ratas Endogámicas Lew , Bazo/efectos de los fármacos , Bazo/inmunología , Vagina/inmunología , Vagina/microbiología , Vagina/patologíaRESUMEN
PROBLEM: To determine whether cells in the female reproductive tract (FRT) are functionally capable of presenting antigen to T cells. METHOD OF STUDY: Analysis was done by determining the proliferation of purified autologous T cells to antigen, following co-incubation with non-proliferating cell suspensions isolated from the uterus and prepared by enzymatic digestion of reproductive tract tissues from hysterectomy patients with benign disease. RESULTS: All uterine preparations analyzed were functionally capable of presenting antigen; the ability to present antigen was independent of pre- and post-menopausal status. In contrast, some, but not all, tissues from the ovary, Fallopian tube, cervix, and vagina were capable of presenting antigen. CONCLUSION: These results suggest that the human FRT is an inductive site for immune responses. Regulation of antigen presentation in the reproductive tract may be important for protection against sexually transmitted diseases.
Asunto(s)
Presentación de Antígeno , Células Presentadoras de Antígenos , Posmenopausia/inmunología , Premenopausia/inmunología , Femenino , Genitales Femeninos/inmunología , Humanos , Inmunidad Mucosa , Activación de Linfocitos , Linfocitos T/efectos de los fármacos , Toxoide Tetánico/farmacología , Útero/citología , Útero/inmunologíaRESUMEN
To determine the influence of human uterine cells recovered at different stages of the menstrual cycle and following menopause on the proliferation of peripheral blood mononuclear cells (PBMC), whole cell suspensions of uterine tissues were co-cultured with autologous and heterologous PBMC. PBMC proliferation in response to tetanus toxoid (TT) or Con A was inhibited by uterine endometrial cells and was dependent on the phase of the menstrual cycle. Inhibition by cells from the proliferative phase was significantly greater than by cells from the secretory phase. Uterine cells isolated from post-menopausal women also inhibited proliferation of PBMC. Cell fractionation studies indicated that epithelial cells are the primary source of uterine inhibitory activity. When epithelial cells and PBMC were cultured in separate compartments, epithelial cells released a soluble factor(s) that inhibited the PBMC proliferation. These results suggest that uterine epithelial cells produce cytokines that down-regulate the proliferation of PBMC in response to antigens and mitogens. This may be important for the control of uterine immune responses, as well as the growth of the reproductive tract in preparation for implantation during the secretory phase of the menstrual cycle.