RESUMEN
The maternal-fetal interface undergoes dynamic changes to allow the fetus to grow and develop in the uterus, despite being recognized by the maternal immune cells. Within the innate immune system, decidual natural killer cells and antigen presenting cells (including macrophages and dendritic cells) that comprise a large proportion of the decidual leukocyte populations play an important role in modulating trophoblast invasion, angiogenesis and vascular remodeling. On the other hand, within the adaptive immune system, CD8+ T cells, effector CD4+ T cells, Foxp3+ regulatory T cells and CD4+HLA-G+ suppressor T cells are identified as potential players in maintaining immune tolerance toward the semi-allogeneic fetus. This review discusses how these key immune cells contribute to pregnancy outcome and the complex interactions between the innate and adaptive immune system during human pregnancy.
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Células Presentadoras de Antígenos/inmunología , Decidua/inmunología , Células Asesinas Naturales/inmunología , Linfocitos T Reguladores/inmunología , Inmunidad Adaptativa , Animales , Decidua/patología , Femenino , Factores de Transcripción Forkhead/metabolismo , Homeostasis , Humanos , Tolerancia Inmunológica , Inmunidad Innata , Circulación Placentaria , EmbarazoRESUMEN
Circulating microRNAs (miRNAs) have emerged as candidate biomarkers of various diseases and conditions including malignancy and pregnancy. This approach requires sensitive and accurate quantitation of miRNA concentrations in body fluids. Herein we report that enzyme-based miRNA quantitation, which is currently the mainstream approach for identifying differences in miRNA abundance among samples, is skewed by endogenous serum factors that co-purify with miRNAs and anticoagulant agents used during collection. Of importance, different miRNAs were affected to varying extent among patient samples. By developing measures to overcome these interfering activities, we increased the accuracy, and improved the sensitivity of miRNA detection up to 30-fold. Overall, the present study outlines key factors that prevent accurate miRNA quantitation in body fluids and provides approaches that enable faithful quantitation of miRNA abundance in body fluids.
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Biomarcadores de Tumor/sangre , MicroARNs/sangre , Neoplasias/sangre , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Anticoagulantes/química , Células Cultivadas , ARN Polimerasas Dirigidas por ADN/antagonistas & inhibidores , ARN Polimerasas Dirigidas por ADN/química , Liasa de Heparina/química , Humanos , Neoplasias/diagnóstico , Oxalatos/química , Plasma/química , Estabilidad del ARN , Sensibilidad y Especificidad , Fluoruro de Sodio/química , Manejo de EspecímenesRESUMEN
Cyclical hormonal changes during an ovarian cycle may affect immune responses, which is crucial for the embryonic implantation. We aim to investigate whether the levels and activity of T, B, and NK cells change during a menstrual cycle. Twenty-two normally cycling women were enrolled and peripheral blood was drawn serially during a menstrual cycle. Intracellular cytokine expression of CD3(+)CD4(+) and CD3(+)CD8(+) cells, and Th1/Th2 cytokine-producing T cell ratios were determined using flow cytometric analysis. NK cell cytotoxicity was measured by flow cytometric analysis at E:T ratios of 50:1, 25:1, and 12.5:1 and also using LU at 20%. Proportions (percentage) of CD3(+) (p = 0.046) and CD3(+)CD4(+) (p = 0.002) T cells were increased in the follicular phase compared with the luteal phase. The levels of CD3(-)CD56(+) (p = 0.010) and CD3(-)CD56(dim) (p = 0.012) NK cells and NK cytotoxicity at E:T ratio of 50:1, 25:1, and 12.5:1 and LU at 20% were significantly increased in the luteal phase compared with the follicular phase. Even though IL-10-producing CD3(+)CD4(+) T cells were significantly lower in the midluteal phase as compared with the early follicular phase, proportions of CD19(+) B cells, CD3(+)CD56(+) NKT cells, Th1 cytokine-producing T cell subsets, and ratios of Th1/Th2 cytokine-producing T cells were not significantly changed during a menstrual cycle. We conclude that peripheral blood NK and T cell levels as well as NK cytotoxicity are changed during a menstrual cycle. Neuroendocrine regulation on immune responses is suggested during an ovarian cycle, which may be critical for embryonic implantation and pregnancy.
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Subgrupos de Linfocitos B/citología , Subgrupos de Linfocitos B/inmunología , Células Asesinas Naturales/citología , Células Asesinas Naturales/inmunología , Ciclo Menstrual/inmunología , Subgrupos de Linfocitos T/citología , Subgrupos de Linfocitos T/inmunología , Adulto , Subgrupos de Linfocitos B/metabolismo , Estudios Transversales , Citocinas/biosíntesis , Citocinas/sangre , Pruebas Inmunológicas de Citotoxicidad , Femenino , Humanos , Inmunofenotipificación , Recuento de Linfocitos , Estudios Prospectivos , Subgrupos de Linfocitos T/metabolismo , Células TH1/citología , Células TH1/inmunología , Células TH1/metabolismo , Células Th2/citología , Células Th2/inmunología , Células Th2/metabolismo , Salud de la Mujer , Adulto JovenRESUMEN
PROBLEM: Women with antiphospholipid antibodies (aPL) are at risk of recurrent miscarriage and pre-eclampsia. aPL target the placenta by binding to beta(2)-glycoprotein I (beta(2) GPI) expressed by the trophoblast. The objective of this study was to evaluate if and how aPL affect first trimester trophoblast migration. METHOD OF STUDY: First trimester trophoblast cells were treated with anti-beta(2) GPI monoclonal antibodies. Migration was determined using a two-chamber assay. Interleukin (IL)-6 production was evaluated by RT-PCR and enzyme-linked immunosorbent assay, and signal transducer and activator of transcription 3 (STAT3) activation was assessed by western blot. RESULTS: Trophoblast cells constitutively secreted IL-6 in a time-dependent manner and this directly correlated with STAT3 phosphorylation. In the presence of anti-beta(2) GPI Abs, trophoblast IL-6 mRNA levels and secretion was downregulated in a Toll-like receptor 4/MyD88-independent manner and this correlated with a reduction in phosphorylated STAT3 levels. In addition, the anti-beta(2) GPI Abs reduced the migratory potential of trophoblast. Heparin was able to reverse aPL-dependent inhibition of trophoblast IL-6 secretion and migration. CONCLUSION: This study demonstrates that aPL limit trophoblast cell migration by downregulating trophoblast IL-6 secretion and STAT3 activity. As heparin was unable to prevent these effects, our findings may explain why women with antiphospholipid syndrome, treated with heparin, remain at risk of developing obstetrical syndromes, associated with impaired deep placentation, such as pre-eclampsia.
Asunto(s)
Anticuerpos Antifosfolípidos/inmunología , Movimiento Celular , Regulación hacia Abajo , Interleucina-6/biosíntesis , Factor de Transcripción STAT3/metabolismo , Trofoblastos/citología , Trofoblastos/inmunología , Células Cultivadas , Humanos , Interleucina-6/inmunología , Interleucina-6/metabolismo , Fosforilación , Transducción de Señal , Trofoblastos/metabolismo , beta 2 Glicoproteína I/inmunologíaRESUMEN
It is becoming clear that during each developmental stage of pregnancy, different immunological conditions exist and may even be necessary for success. The widely accepted T helper (Th) 1 and 2 concept has some limitations if applied to the various developmental stages of pregnancy. During the implantation period, a multidirectional cytokine network is necessary with the blastocyst producing cytokines and other factors and the endometrium synthesizing factors necessary for the embryonic development. Improper immune responses and an unbalanced cytokine network may be related to implantation failures, pregnancy losses and obstetrical complications. A propensity to Th1 immune responses has been reported in these conditions systemically or locally. The presence of elevated Th1:Th2 cell ratios, high concentration of Th1 cytokines secreted by peripheral blood mononuclear cells, elevated NK cell cytotoxicity and levels, and emergence of various autoantibodies are the supporting evidence. The underlying immunopathology for the preponderance of Th1 is unknown. Genetic, environmental, and hormonal etiologies need to be explored further in the future. The purpose of this review is to give an overview of what is known about the immune response in women with reproductive failures and provide an update of some of the most recent findings in this field.