RESUMEN
Erythema Toxicum, a rash frequently present in the healthy newborn infant is an innate, immune response to the first commensal micro flora. Flushing and urtication are seen in this manifestation suggesting mast cell (MC) activation and MC derived mediator release. It has recently become evident that MCs participate in the protective, innate immune response against microbes also by secreting products toxic to pathogens such as cathelicidin peptide antibiotics. We hypothesized that MCs contribute to the process of inflammation in Erythema Toxicum and that skin MCs of human newborns express the cathelicidin peptide antibiotic LL-37. Skin sections were immunostained for MC tryptase. Double immunofluorescence was performed by staining LL-37 in combination with tryptase. We studied ultra structure of skin MCs with transmission (TEM) and immunoelectron microscopy (IEM). Seven infants with and six infants without the rash, as well as three adults were included. We found numerous tryptase-expressing MCs recruited around the hair follicles in the lesions of Erythema Toxicum. TEM analysis of MCs exhibited signs of degranulation in the lesion. Neither skin MCs from newborns nor adults did express LL-37 as judged by confocal and IEM. MCs participate in the inflammatory responses of Erythema Toxicum by taking an active part in the immune system of the hair follicle. However, their immunological activity is not linked to the expression of the cathelicidin antimicrobial peptide LL-37. A pivotal role of MCs in the innate, inflammatory response at the site of pathogen invasion during the critical time of perinatal colonization is suggested.
Asunto(s)
Eritema/inmunología , Folículo Piloso/inmunología , Mastocitos/inmunología , Piel/inmunología , Triptasas/metabolismo , Urticaria/inmunología , Adulto , Péptidos Catiónicos Antimicrobianos/análisis , Catelicidinas , Eritema/metabolismo , Femenino , Humanos , Recién Nacido , Masculino , Mastocitos/enzimología , Mastocitos/ultraestructura , Microscopía Electrónica de Transmisión , Microscopía Inmunoelectrónica , Piel/metabolismo , Piel/ultraestructura , Urticaria/metabolismoRESUMEN
At birth, commensal microbes penetrate into the skin of the human newborn, eliciting an acute rash, erythema toxicumn neonatorum. Histologically, the rash is characterized by an upregulation of proinflammatory activity and a local recruitment of immunocytes, including macrophages. High mobility group box chromosomal protein 1, a nuclear and cytosolic protein, is also a pro-inflammatory cytokine released by macrophages in response to microbial stimulation. Here, we reasoned that macrophages but also keratinocytes might upregulate this protein in response to the first colonization and that high mobility group box chromosomal protein 1 might play a role as a proinflammatory mediator in the development and progression of erythema toxicum. Punch biopsy specimens from 1-day-old healthy infants, seven with and four without erythema toxicum were analyzed with indirect immunohistochemistry and two different antihigh mobility group box chromosomal protein 1 antibodies, immunofluorescence, nuclear counterstaining, confocal and immunoelectron imaging. We found relocation of nuclear high mobility group box chromosomal protein 1 into the cytoplasm in keratinocytes and macrophages in erythema toxicum. Cytoplasmatic high mobility group box chromosomal protein 1 was also found in melanocytes and did neither co-locate with lysosomal-associated membrane proteins nor with melanosomes. We speculate that terrestrial adaptation triggers the induction of the endogenous "danger signal" high mobility group box chromosomal protein 1 in the skin of the newborn infant, perhaps in response to the first commensal colonization and that this signal may contribute to alert the immune system and promote a protective immune response.
Asunto(s)
Eritema/patología , Proteína HMGB1/metabolismo , Enfermedades del Recién Nacido/patología , Queratinocitos/metabolismo , Piel/patología , Bacterias/inmunología , Biopsia , Eritema/inmunología , Eritema/microbiología , Técnica del Anticuerpo Fluorescente , Humanos , Recién Nacido , Enfermedades del Recién Nacido/inmunología , Enfermedades del Recién Nacido/microbiología , Queratinocitos/patología , Queratinocitos/ultraestructura , Proteína 2 de la Membrana Asociada a los Lisosomas , Proteínas de Membrana de los Lisosomas/metabolismo , Macrófagos/metabolismo , Macrófagos/patología , Macrófagos/ultraestructura , Proteínas de la Membrana/metabolismo , Microscopía Confocal , Microscopía Inmunoelectrónica , Piel/inmunología , Piel/microbiología , Proteínas de Transporte Vesicular/metabolismoRESUMEN
OBJECTIVE: To study the effect of mifepristone on heparin-binding epidermal growth factor (HB-EGF) and its receptors HER1 and HER4 in the fallopian tube and in the endometrium. DESIGN: Prospective clinical study. SETTING: Hospital-based unit for obstetrics and gynecology and research laboratories. PATIENT(S): Healthy women divided into two groups: controls and patients treated with a single dose of 200 mg mifepristone on day LH+2. INTERVENTION(S): Endometrial biopsies from 30 women were obtained during one control cycle or one treatment cycle. Fallopian tubes from 14 women were collected during laparoscopic sterilizations. MAIN OUTCOME MEASURE(S): Immunohistochemistry and reverse transcriptase-polymerase chain reaction. RESULT(S): The staining intensity of HB-EGF was not affected by mifepristone treatment. Treatment with mifepristone increased the immunostaining on HER1 in the epithelium and the stroma of the endometrium, which was not seen in the fallopian tube. The immunostaining of HER4 decreased in the stroma of the fallopian tube, while an increase was seen in the epithelial cells of the endometrium. CONCLUSION(S): Treatment with mifepristone has a limited effect on HB-EGF and its receptors in the fallopian tube, while the increase in HER1 and HER4 in the endometrium probably reflects defective endometrial maturation.
Asunto(s)
Anticonceptivos Sintéticos Orales/administración & dosificación , Factor de Crecimiento Epidérmico/genética , Receptores ErbB/genética , Mifepristona/administración & dosificación , Adulto , Endometrio/efectos de los fármacos , Endometrio/fisiología , Factor de Crecimiento Epidérmico/metabolismo , Receptores ErbB/metabolismo , Trompas Uterinas/efectos de los fármacos , Trompas Uterinas/fisiología , Femenino , Expresión Génica/efectos de los fármacos , Factor de Crecimiento Similar a EGF de Unión a Heparina , Humanos , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intercelular , Laparoscopía , Persona de Mediana Edad , Estudios Prospectivos , Receptor ErbB-4 , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Esterilización ReproductivaRESUMEN
OBJECTIVES: The objective of the present study was to investigate the effect of vaginally and orally administered misoprostol on the local cervical inflammatory response. METHODS: Healthy women with a normal intrauterine pregnancy between 8 and 12 weeks of gestation presenting for an elective termination of pregnancy by vacuum aspiration were recruited into a cohort study with a control and a treatment group. In the treatment group, the women were randomized to misoprostol, 400 mug, given either orally or vaginally 3 h before surgery. Immunohistochemistry staining of CD45, CD68, MMP 8, MMP 9, TIMP 1 and TIMP 2 were assessed in cervical biopsies obtained directly prior to mechanical cervical dilatation and vacuum aspiration. RESULTS: In the treatment group, there was a greater amount of CD45-positive cells in the subepithelium region of the cervix compared to the control group. The staining of CD68 was similar in both groups. The immunostaining of MMP 8 and MMP 9 was greater in the treatment group, while the expression of TIMP 1 and TIMP 2 did not differ between control and treatment groups. CONCLUSIONS: Compared to untreated controls, treatment with misoprostol was associated with a greater expression of inflammatory cells. It could be hypothesized that administration of misoprostol mimics the cervical ripening at term pregnancy by a possible influx and activation of inflammatory cells, which increases MMP 8 and MMP 9 and thereby leads to the degradation of collagen and cervical softening.
Asunto(s)
Abortivos no Esteroideos/administración & dosificación , Maduración Cervical/efectos de los fármacos , Edad Gestacional , Inflamación , Misoprostol/administración & dosificación , Aborto Inducido , Administración Intravaginal , Administración Oral , Adulto , Antígenos CD/análisis , Antígenos de Diferenciación Mielomonocítica/análisis , Femenino , Humanos , Inmunohistoquímica , Antígenos Comunes de Leucocito/análisis , Metaloproteinasa 8 de la Matriz/análisis , Metaloproteinasa 9 de la Matriz/análisis , Embarazo , Inhibidor Tisular de Metaloproteinasa-1/análisis , Inhibidor Tisular de Metaloproteinasa-2/análisis , Legrado por AspiraciónRESUMEN
We have examined the cellular localization and human amniotic fluid content of endothelin-1 (ET-1) and macrophage colony-stimulating factor (M-CSF). The study material consisted of amniotic fluid from 20 patients referred for amniocentesis, and placental samples from normal deliveries. ET-1 and M-CSF were analysed by radioimmunoassay and enzyme-linked immunosorbent assay respectively. The cellular localization of ET-1 and M-CSF in the amnion membranes was analysed by double-labelling immunocytochemistry using fluorescein isothiocyanate- and Cy3-labelled secondary antibodies. Release of ET-1 and M-CSF was studied in cultured amniocytes. We found that the mean +/- SD concentrations of ET-1 and M-CSF in fetal amniotic fluid were 45.6 +/- 17.3 pmol/l (range 16.8-85.5) and 7323 +/- 3415 ng/l (range 2640-12 110) respectively. Double-labelling immunocytochemistry showed that both M-CSF and ET-1 were co-localized in the same cells to a high extent. Further analysis revealed that levels of M-CSF, but not ET-1, were significantly correlated with pregnancy length. Both M-CSF and ET-1 were released from cultured amniocytes in response to interleukin-1. These findings show that ET-1 and M-CSF are partly co-localized to specific cells in the human amniotic membrane. As both M-CSF and ET-1 were released from cultured amniocytes in vitro, this suggests that they both may be secreted into fetal amniotic fluid in vivo as well.
Asunto(s)
Amnios/química , Líquido Amniótico/química , Endotelina-1/análisis , Endotelina-1/metabolismo , Factor Estimulante de Colonias de Macrófagos/análisis , Factor Estimulante de Colonias de Macrófagos/metabolismo , Adolescente , Adulto , Amnios/citología , Líquido Amniótico/citología , Peso al Nacer , Desarrollo Embrionario y Fetal , Femenino , Humanos , Embarazo , Trimestres del EmbarazoRESUMEN
Erythema toxicum neonatorum is a common, inflammatory skin reaction in healthy newborn infants characterized by an accumulation of activated immune cells in the lesions. Its etiology and physiologic significance are still unclear. The purpose of this study was to extend the search for possible inflammatory mediators of the rash. We performed immunohistochemistry on punch biopsy cryosections from lesions of four, 1-day-old infants and from four matched controls without rash, using antibodies against the water channel proteins aquaporin-1 (AQP1) and aquaporin-3 (AQP3), psoriasin, and the nitric oxide synthase (NOS) enzymes, neuronal NOS (nNOS), inducible NOS (iNOS), and endothelial NOS (eNOS). All sections from the lesions showed a dense, nodular cellular infiltrate located near the hair follicle. The vessels in the dermis showed a high incidence of AQP1 and eNOS. Strong staining for AQP1, AQP3, and psoriasin, as well as nNOS, iNOS, and eNOS were seen in the entire epidermal layer. The infiltrate in the dermis contained numerous cells expressing AQP1, AQP3, nNOS, iNOS, and eNOS. Double immunofluorescence staining showed that AQP3 was located in CD1a-expressing Langerhans cells and other dendritic cells in the dermis, as well as in CD14-expressing macrophages, CD15-expressing neutrophils, and EG2-expressing eosinophils surrounding the hair follicle. Our findings show that AQP1 and AQP3, psoriasin, and NOSs are involved in the activation of the skin immune system at birth.
Asunto(s)
Acuaporinas/metabolismo , Proteínas de Unión al Calcio/metabolismo , Eritema/metabolismo , Mediadores de Inflamación/metabolismo , Óxido Nítrico Sintasa/metabolismo , Piel/química , Acuaporina 1 , Acuaporina 3 , Antígenos de Grupos Sanguíneos , Eritema/patología , Femenino , Humanos , Inmunohistoquímica , Recién Nacido , Masculino , Proteína A7 de Unión a Calcio de la Familia S100 , Proteínas S100 , Piel/patologíaRESUMEN
OBJECTIVE: To investigate the effects of the antiprogestin mifepristone on the expression of insulin-like growth factor binding protein-1 (IGFBP-1) mRNA and protein during the early luteal phase in the human oviduct. DESIGN: Prospective case-control study. SETTING: University hospital. PATIENT(S): Fourteen healthy women with regular menstrual cycles who were admitted to the hospital for voluntary sterilization by the laparoscopic technique. INTERVENTION(S): Treatment with 200 mg of mifepristone was administered on day LH+2. Fallopian tube samples were obtained on days LH+4 to LH+6. MAIN OUTCOME MEASURE(S): Expression of IGFBP-1 was identified using immunhistochemistry, and mRNA levels were determined with semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). RESULT(S): Immunoreactivity for IGFBP-1 was primarily localized to the cytoplasm of the oviductal epithelial cells. Messenger RNA for IGFBP-1 was identified in total RNA extracted from the same fallopian tube samples. There was a significant increase in the expression of IGFBP-1 immunostaining and mRNA after treatment with mifepristone. CONCLUSION(S): These data further illustrate the complex actions of mifepristone and support the view that changes in the oviductal environment after treatment with mifepristone may be detrimental to normal gamete transport and function and contribute to the contraceptive action of mifepristone.
Asunto(s)
Trompas Uterinas/metabolismo , Antagonistas de Hormonas/farmacología , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina/biosíntesis , Fase Luteínica/metabolismo , Mifepristona/farmacología , Adulto , Estudios de Casos y Controles , Trompas Uterinas/efectos de los fármacos , Femenino , Humanos , Inmunohistoquímica , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Fase Luteínica/efectos de los fármacos , Persona de Mediana Edad , Estudios Prospectivos , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Insulin-like growth factor binding protein-1 (IGFBP-1) and prolactin are recognized as crucial signals for the initiation and maintainance of decidualization. The purpose of the study was to investigate the effect of mifepristone on the expression of IGFBP-1, prolactin and progesterone receptors (PR) during the implantation phase in human endometrium. Eight fertile women were studied during control and treatment cycles. Treatment with 200 mg of mifepristone was administered on day LH +2. Endometrial samples were collected on day LH +6 to +8. Expression of IGFBP-1, prolactin and PR was identified using immunohistochemistry, and mRNA levels were determined with RT-PCR. In control specimens, IGFBP-1 and prolactin were localized to the cytoplasm of the endometrial glandular and to a lesser extent in stromal cells. In the same samples, PR immunoreactivity was detected in the nucleus of the endometrial stromal cells, and was absent from the glandular cells. After mifepristone treatment, there was a significant increase in the immunostaining and mRNA expression for IGFBP-1 and PR. Prolactin expression increased only slightly after treatment. These results support the view that administration of mifepristone in the early luteal phase does not simply retard endometrial development. Our findings provide further insight into the regulation of IGFBP-1 and prolactin by PR in the human endometrium in vivo.
Asunto(s)
Implantación del Embrión/efectos de los fármacos , Endometrio/efectos de los fármacos , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Mifepristona/farmacología , Prolactina/genética , Receptores de Progesterona/genética , Abortivos Esteroideos/farmacología , Adulto , Implantación del Embrión/fisiología , Endometrio/fisiología , Femenino , Humanos , Inmunohistoquímica/métodos , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina/efectos de los fármacos , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Prolactina/efectos de los fármacos , Prolactina/metabolismo , ARN Mensajero/efectos de los fármacos , Receptores de Progesterona/efectos de los fármacos , Receptores de Progesterona/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: To study the effect of mifepristone and levonorgestrel on ovarian function and endometrial development in doses effective as emergency contraception. METHODS: Twelve fertile women were treated with either 10 mg of mifepristone as a single dose (n = 6) or two doses of 0.75 mg of levonorgestrel, 12 hours apart (n = 6) before and after ovulation. An endometrial biopsy performed during the implantation period was analyzed for endometrial maturation and expression of markers of endometrial receptivity. The markers tested for were integrin alpha4 and beta3, cyclooxygenase-1 and -2, progesterone receptors, Dolichos biflorus agglutinin lectin binding, and pinopodes. Urinary excretion of luteinizing hormone, estrone, and pregnanediol were also determined. RESULTS: Treatment with mifepristone and levonorgestrel before ovulation inhibited the luteinizing hormone surge showing no significant differences between the means of luteinizing hormone measurements. When mifepristone was administered in the early luteal phase, downregulation of progesterone receptors was inhibited in five of six women. No significant alteration was found in any of the remaining markers of endometrial receptivity. CONCLUSION: The mode of action of emergency contraception with mifepristone or levonorgestrel is primarily due to inhibition of ovulation rather than inhibition of implantation.