RESUMEN

PURPOSE: Incisional hernia occurs in up to 20% of patients after abdominal surgery and is most common after vertical midline incisions. Diastasis recti may contribute to incisional hernia but has not been explored as a risk factor or included in hernia risk models. We examined the association between diastasis recti and incisional hernia after midline incisions. METHODS: In this single-center study, all patients undergoing elective gastrointestinal surgery with a midline open incision or extraction site in a prospective surgical quality collaborative database between 2016 and 2020 were included. Eligible patients had axial imaging within 6 months prior to surgery and no less than 6 months after surgery to determine the presence of diastasis recti and incisional hernia, respectively. Radiographic hernia-free survival was assessed with log-rank tests and multivariable Cox regression, comparing patients with and without diastasis width > 25 mm. RESULTS: Of 156 patients, forty-four (28.2%) developed radiographic hernia > 1 cm. 36 of 85 patients (42.4%) with DR width > 25 mm developed IH, compared to 9 of 71 (12.7%) without DR (p < 0.001). Hernia-free survival differed by DR width on bivariate and multivariable Cox regression, adjusted hazard ratio: 3.87, 95% confidence interval: 1.84-8.14. CONCLUSION: Diastasis recti is a significant risk factor for incisional hernia after midline abdominal surgery. When present, surgeons can include these data when discussing surgical risks and should consider a lower risk, off-midline approach when feasible. Incorporating diastasis into larger studies may improve comprehensive models of incisional hernia risk.

Asunto(s)

Procedimientos Quirúrgicos del Sistema Digestivo , Hernia Ventral , Hernia Incisional , Humanos , Hernia Incisional/cirugía , Hernia Ventral/cirugía , Estudios Prospectivos , Herniorrafia/efectos adversos

RESUMEN

BACKGROUND: Uterine serous papillary carcinoma (USPC) is a biologically aggressive variant of endometrial cancer. We investigated the expression of Serum Amyloid A (SAA) and evaluated its potential as a serum biomarker in USPC patients. METHODS: SAA gene and protein expression levels were evaluated in USPC and normal endometrial tissues (NEC) by real-time PCR, immunohistochemistry (IHC), flow cytometry and by a sensitive bead-based immunoassay. SAA concentration in 123 serum samples from 51 healthy women, 42 women with benign diseases, and 30 USPC patients were also studied. RESULTS: SAA gene expression levels were significantly higher in USPC when compared with NEC (mean copy number by RT-PCR=162 vs 2.21; P=0.0002). IHC revealed diffuse cytoplasmic SAA protein staining in USPC tissues. High intracellular levels of SAA were identified in primary USPC cell lines evaluated by flow cytometry and SAA was found to be actively secreted in vitro. SAA concentrations (mug ml(-1)) had a median (95% CIs) of 6.0 (4.0-8.9) in normal healthy females and 6.0 (4.2-8.1) in patients with benign disease (P=0.92). In contrast, SAA values in the serum of USPC patients had a median (95% CI) of 15.6 (9.2-56.2), significantly higher than those in the healthy group (P=0.0005) and benign group (P=0.0006). Receiver operating characteristics (ROC) analysis of serum SAA to classify advanced- and early-stage USPC yielded an area under the ROC curve of 0.837 (P=0.0024). CONCLUSION: SAA is not only a liver-secreted protein but is also a USPC cell product. SAA may represent a novel biomarker for USPC to assist in staging patients preoperatively, and to monitor early-disease recurrence and response to therapy.

Asunto(s)

Biomarcadores de Tumor/sangre , Carcinoma Papilar/sangre , Cistadenocarcinoma Seroso/sangre , Proteína Amiloide A Sérica/biosíntesis , Neoplasias Uterinas/sangre , Biomarcadores de Tumor/biosíntesis , Biomarcadores de Tumor/genética , Carcinoma Papilar/genética , Carcinoma Papilar/patología , Cistadenocarcinoma Seroso/genética , Cistadenocarcinoma Seroso/patología , Femenino , Citometría de Flujo , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Estadificación de Neoplasias , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteína Amiloide A Sérica/genética , Células Tumorales Cultivadas , Neoplasias Uterinas/genética , Neoplasias Uterinas/patología

RESUMEN

Cancer progression is an abnormal form of tissue repair characterized by chronic inflammation. IkappaB kinase-beta (IKKbeta) required for nuclear factor-kappaB (NF-kappaB) activation plays a critical role in this process. Using EOC cells isolated from malignant ovarian cancer ascites and solid tumors, we identified IKKbeta as a major factor promoting a functional TLR-MyD88-NF-kappaB pathway that confers to EOC cell the capacity to constitutively secrete proinflammatory/protumor cytokines and therefore promoting tumor progression and chemoresistance. Furthermore, we describe for the first time the identification of the microRNA hsa-miR-199a as a regulator of IKKbeta expression. Our study describes the property of ovarian cancer cells to enhance the inflammatory microenvironment as a result of the expression of an active IKKbeta pathway. Identification of these markers in patients' tumor samples may facilitate the adequate selection of treatment and open new venues for the development of effective therapy for chemoresistant ovarian cancers.

Asunto(s)

Quinasa I-kappa B/genética , MicroARNs/fisiología , FN-kappa B/metabolismo , Neoplasias Ováricas/genética , Neoplasias Ováricas/metabolismo , Secuencia de Bases , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Modelos Biológicos , Datos de Secuencia Molecular , Homología de Secuencia de Ácido Nucleico , Transfección , Células Tumorales Cultivadas

RESUMEN

PROBLEM: Macrophages are one of the first immune cells observed at the implantation site. Their presence has been explained as the result of an immune response toward paternal antigens. The mechanisms regulating monocyte migration and differentiation at the implantation site are largely unknown. In the present study, we demonstrate that trophoblast cells regulate monocyte migration and differentiation. We propose that trophoblast cells 'educate' monocytes/macrophages to create an adequate environment that promote trophoblast survival. METHOD OF STUDY: CD14(+) monocytes were isolated from peripheral blood using magnetic beads. Co-culture experiments were conducted using a two-chamber system. Monocytes were stimulated with lipopolysaccharide (LPS) and cytokine levels were determined using multiplex cytokine detecting assay. RESULTS: Trophoblast cells increase monocyte migration and induce a significant increase in the secretion and production of the pro-inflammatory cytokines [interleukin-6 (IL-6), IL-8, tumor necrosis factor-alpha] and chemokines (growth-related oncogen-alpha, monocyte chemoattractant protein-1, macrophage inflammatory protein-1beta, RANTES). Furthermore, the response of monocytes to LPS was different in monocytes pre-exposed to trophoblast cells. CONCLUSION: The results of this study suggest that trophoblast cells are able to recruit and successfully educate monocytes to produce and secrete a pro-inflammatory cytokine and chemokine profile supporting its growth and survival. Furthermore we demonstrate that trophoblast cells can modulate monocytes response to bacterial stimuli.

Asunto(s)

Comunicación Celular , Macrófagos/citología , Trofoblastos/citología , Línea Celular , Movimiento Celular , Técnicas de Cocultivo , Citocinas/biosíntesis , Citocinas/metabolismo , Femenino , Humanos , Receptores de Lipopolisacáridos/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/metabolismo , Monocitos/citología , Monocitos/efectos de los fármacos , Monocitos/metabolismo , Embarazo , Trofoblastos/metabolismo

RESUMEN

Activated CD4 Th1 lymphocytes can enter the brain in the absence of an inflammatory focus. However, the molecular mediators that regulate this early migration of lymphocytes into the brain have remained unclear. We hypothesized that the entry of these 'pioneer' lymphocytes into the brain is regulated by a set of molecular events that are distinct from those used once inflammation has been established. Using cells fluorescently labelled with the lipophilic dye DiI, myelin basic protein (MBP)-specific CD4 lymphocytes that expressed low or high levels of very late antigen-4 (VLA-4) and non-antigen-specific activated splenocytes homed to mouse brain in similar quantities 2 h after adoptive transfer. However, antigen specificity and VLA-4 expression were required for more robust recruitment by 24 h. Immunocytochemistry revealed endothelial and microenvironmental upregulation of vascular cell adhesion molecule (VCAM), intercellular cell adhesion molecule 1 (ICAM-1), MHC class II and interferon-gamma 48 h after transfer of MBP-specific cells. In contrast, expression of meningeal and choroid plexus-associated P selectin was upregulated 2 h after adoptive transfer, but not at 48 h. Monoclonal antibody to P selectin, but not to VLA-4, inhibited early migration of high VLA-4-expressing MBP-specific lymphocytes. These results suggest that early migration occurs independent of the lymphocyte integrin VLA-4 and endothelial VCAM, but does require increased surface expression of endothelial P selectin.

Asunto(s)

Química Encefálica/inmunología , Encefalomielitis Autoinmune Experimental/inmunología , Vigilancia Inmunológica , Integrinas/inmunología , Receptores Mensajeros de Linfocitos/inmunología , Molécula 1 de Adhesión Celular Vascular/inmunología , Animales , Antialérgicos/inmunología , Antialérgicos/metabolismo , Anticuerpos Monoclonales , Encéfalo/citología , Encéfalo/inmunología , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Carbocianinas , Movimiento Celular/inmunología , Encefalomielitis Autoinmune Experimental/metabolismo , Femenino , Colorantes Fluorescentes , Antígenos de Histocompatibilidad Clase II/inmunología , Integrina alfa4beta1 , Integrinas/metabolismo , Molécula 1 de Adhesión Intercelular/inmunología , Molécula 1 de Adhesión Intercelular/metabolismo , Interferón gamma/inmunología , Ratones , Ratones Endogámicos , Proteína Básica de Mielina/inmunología , Selectina-P/inmunología , Páncreas/citología , Páncreas/inmunología , Receptores Mensajeros de Linfocitos/metabolismo , Bazo/citología , Células TH1/citología , Células TH1/inmunología , Molécula 1 de Adhesión Celular Vascular/metabolismo

RESUMEN

Type 1 diabetes is an autoimmune disease in which the insulin-producing pancreatic beta cells are destroyed at an early age by an immune process that involves both CD4 and CD8 T lymphocytes. The identification of autoantigens in diabetes is very important for the design of antigen-specific immunotherapy. By screening a pancreatic islet cDNA library, we have identified the autoantigen recognized by highly pathogenic CD8 T cells in the non-obese diabetic mouse, one of the best animal models for human diabetes. This is the first identification, to our knowledge, of a CD8 T-cell epitope in an autoimmune disease. The peptide recognized by the cells is in the same region of the insulin B chain as the epitope recognized by previously isolated pathogenic CD4 T cells. This has very important implications for the potential use of insulin in preventative therapy.

Asunto(s)

Autoantígenos/inmunología , Linfocitos T CD8-positivos/inmunología , Diabetes Mellitus Tipo 1/inmunología , Biblioteca de Genes , Antígenos de Histocompatibilidad Clase I/inmunología , Islotes Pancreáticos/inmunología , Secuencia de Aminoácidos , Animales , Linfocitos T CD8-positivos/patología , Células COS , Células Clonales/inmunología , Células Clonales/patología , Clonación Molecular , Diabetes Mellitus Tipo 1/genética , Epítopos de Linfocito T/química , Epítopos de Linfocito T/genética , Epítopos de Linfocito T/inmunología , Insulina/química , Insulina/genética , Insulina/inmunología , Interferón gamma/biosíntesis , Islotes Pancreáticos/metabolismo , Islotes Pancreáticos/patología , Activación de Linfocitos , Recuento de Linfocitos , Ratones , Ratones Endogámicos NOD , Ratones Endogámicos , Especificidad de Órganos , Péptidos/química , Péptidos/genética , Péptidos/inmunología

RESUMEN

Activated Th1 CD4 T cells bind to P-selectin and migrate into inflamed tissue, whereas Th2 cells do not. We show that alpha(1, 3)-fucosyltransferase VII (FucT-VII) and alpha(2, 3)-sialyltransferase IV (ST3GalIV), which are crucial for the biosynthesis of functional P-selectin ligands, are absent in naive CD4 T cells, but are rapidly up-regulated upon activation. Th1 or Th2 differentiation in the presence of polarizing cytokines leads to down-regulation of FucT-VII mRNA selectively in Th2 but not in Th1 cells. Influencing the differentiation by varying the priming dose of antigenic peptide results in similar FucT-VII down-regulation only in Ag-specific Th2 cells. ST3GalIV levels remain elevated. FucT-VII and ST3GalIV mRNAs are also up-regulated by Th1 cells primed in vivo and recruited into the lymph nodes draining delayed-type hypersensitivity sites. We identify FucT-VII gene expression as a principal difference between Th1 and Th2 cells, and underscore the importance of FucT-VII and ST3GalIV expression for the biosynthesis of functional selectin ligands.

Asunto(s)

Linfocitos T CD4-Positivos/enzimología , Movimiento Celular/inmunología , Fucosiltransferasas/biosíntesis , Ganglios Linfáticos/patología , Activación de Linfocitos , Sialiltransferasas/biosíntesis , Células TH1/enzimología , Animales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/patología , Diferenciación Celular/inmunología , Regulación hacia Abajo/inmunología , Epítopos de Linfocito T/biosíntesis , Fucosiltransferasas/genética , Gangliósidos/inmunología , Antígenos de Histocompatibilidad Clase II/metabolismo , Hipersensibilidad Tardía/enzimología , Hipersensibilidad Tardía/inmunología , Hipersensibilidad Tardía/patología , Interleucina-12/farmacología , Interleucina-4/farmacología , Interfase/inmunología , Antígenos del Grupo Sanguíneo de Lewis/inmunología , Ganglios Linfáticos/enzimología , Ganglios Linfáticos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Ratones Transgénicos , Fragmentos de Péptidos/inmunología , Fragmentos de Péptidos/metabolismo , ARN Mensajero/biosíntesis , Antígeno Sialil Lewis X , Células TH1/inmunología , Células TH1/patología , Células Th2/enzimología , Células Th2/inmunología , Regulación hacia Arriba/inmunología , beta-Galactosida alfa-2,3-Sialiltransferasa

RESUMEN

Bone marrow (BM)-derived dendritic cells (DC) are potent stimulators of naive CD4+ T cell activation. Because DC are efficient at Ag processing and could potentially present self Ags, we investigated the role of DC in the presentation of an encephalitogenic peptide from myelin basic protein (Ac1-11) in the induction of experimental autoimmune encephalomyelitis (EAE). To determine if DC could prime for EAE, we transferred DC pulsed with Ac1-11 or with medium alone into irradiated mice in combination with CD4+ T cells isolated from a mouse transgenic for a TCR specific for Ac1-11 + I-Au. Mice transferred with Ac1-11-pulsed DC developed EAE 7-10 days later, whereas mice receiving medium-pulsed DC did not. By day 15, all mice given peptide-loaded DC had signs of tail and hind limb paralysis, and by day 20 infiltration of Ac1-11-specific CD4+ T cells was detected in the brain parenchyma. We also demonstrated interactions between Ac1-11-pulsed DC and Ac1-11-specific T cells in the lymph nodes 24 h following adoptive transfer of both cell populations. These data show that DC can efficiently present the self Ag myelin basic protein Ac1-11 to Ag-specific T cells in the periphery of mice to induce EAE.

Asunto(s)

Presentación de Antígeno/inmunología , Autoantígenos/metabolismo , Células Dendríticas/inmunología , Encefalomielitis Autoinmune Experimental/etiología , Proteína Básica de Mielina/inmunología , Secuencia de Aminoácidos , Animales , Autoantígenos/genética , Autoantígenos/inmunología , Células de la Médula Ósea/inmunología , Células de la Médula Ósea/metabolismo , Encéfalo/patología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Comunicación Celular/inmunología , Células Clonales , Células Dendríticas/metabolismo , Encefalomielitis Autoinmune Experimental/inmunología , Encefalomielitis Autoinmune Experimental/patología , Epítopos de Linfocito T/inmunología , Epítopos de Linfocito T/metabolismo , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/metabolismo , Activación de Linfocitos , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Datos de Secuencia Molecular , Proteína Básica de Mielina/genética , Proteína Básica de Mielina/metabolismo , Fragmentos de Péptidos/inmunología , Fragmentos de Péptidos/metabolismo , Receptores de Antígenos de Linfocitos T/genética , Médula Espinal/patología , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo , Subgrupos de Linfocitos T/patología , Células TH1/inmunología

RESUMEN

We developed a simple method employing the use of flat-embedding techniques on thick frozen sections which allows correlation of light and electron microscopic immunohistochemistry. This method has been particularly useful in visualization of pancreas sections, an adaptation especially important because this tissue is not amenable to conventional vibratome sectioning. In this study we demonstrate the use of this technique to examine the same tissue section at the light and the electron microscopic level while maintaining morphology.

Asunto(s)

Inmunohistoquímica/métodos , Islotes Pancreáticos/metabolismo , Islotes Pancreáticos/ultraestructura , Microscopía Electrónica/métodos , Animales , Antígenos CD8/metabolismo , Linfocitos T CD8-positivos/metabolismo , Linfocitos T CD8-positivos/ultraestructura , Secciones por Congelación , Insulina/metabolismo , Ratones , Ratones Endogámicos NOD

RESUMEN

B7-1 transgene expression on the pancreatic islets in nonobese diabetic (NOD) mice leads to accelerated diabetes, with >50% of animals developing diabetes before 12 wk of age. The expression of B7-1 directly on the pancreatic beta cells, which do not normally express costimulator molecules, converts the cells into effective antigen-presenting cells leading to an intensified autoimmune attack. The pancreatic islet infiltrate in diabetic mice consists of CD8 T cells, CD4 T cells, and B cells, similar to diabetic nontransgenic NOD mice. To elucidate the relative importance of each of the subsets of cells, the NOD-rat insulin promoter (RIP)-B7-1 animals were crossed with NOD.beta2microglobulin -/- mice which lack major histocompatibility complex class I molecules and are deficient in peripheral CD8 T cells, NOD.CD4 -/- mice which lack T cells expressing CD4, and NOD.muMT -/- mice which lack B220-positive B cells. These experiments showed that both CD4 and CD8 T cells were necessary for the accelerated onset of diabetes, but that B cells, which are needed for diabetes to occur in normal NOD mice, are not required. It is possible that B lymphocytes play an important role in the provision of costimulation in NOD mice which is unnecessary in the NOD-RIP-B7-1 transgenic mice.

Asunto(s)

Antígeno B7-1/biosíntesis , Diabetes Mellitus Tipo 1/inmunología , Islotes Pancreáticos/inmunología , Subgrupos Linfocitarios , Traslado Adoptivo , Edad de Inicio , Animales , Presentación de Antígeno , Antígeno B7-1/genética , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Diabetes Mellitus Tipo 1/etiología , Antígenos de Histocompatibilidad Clase I , Incidencia , Insulina/genética , Islotes Pancreáticos/patología , Ratones , Ratones Endogámicos NOD , Ratones Transgénicos , Regiones Promotoras Genéticas , Bazo/trasplante

RESUMEN

Immunologically privileged sites express Fas ligand (FasL), which protects them from attack by activated T cells that express Fas and die upon contact with FasL. In an attempt to protect nonobese diabetic mice (NOD) from autoimmune diabetes, we made FasL transgenic NOD mice using the beta cell-specific rat insulin-1 promoter. Surprisingly, these transgenic mice showed heightened sensitivity to diabetogenic T cells, which was due to self-destruction of beta cells upon T cell-mediated induction of Fas. Fas-negative NOD(lpr/lpr) animals were resistant to diabetogenic T cells and to spontaneous diabetes. Thus, induction of Fas expression on beta cells and their subsequent destruction constitutes the main pathogenic mechanism in autoimmune diabetes.

Asunto(s)

Diabetes Mellitus Tipo 1/inmunología , Diabetes Mellitus Tipo 1/fisiopatología , Glicoproteínas de Membrana/genética , Animales , Apoptosis/inmunología , Proteína Ligando Fas , Femenino , Citometría de Flujo , Inmunohistoquímica , Insulina/biosíntesis , Islotes Pancreáticos/citología , Islotes Pancreáticos/inmunología , Islotes Pancreáticos/metabolismo , Ligandos , Masculino , Glicoproteínas de Membrana/metabolismo , Ratones , Ratones Endogámicos NOD , Ratones Transgénicos , Regiones Promotoras Genéticas/inmunología , Ratas , Linfocitos T/química , Linfocitos T/inmunología , Transgenes/inmunología

RESUMEN

During inflammation, T cells transmigrate from the bloodstream into perivascular tissues. As T cells transmigrate, they undergo a series of attachments to and detachments from the endothelium and then extravasate into the extracellular matrix (ECM). T cell migration into the ECM involves a number of mechanisms that influence cell-ECM interactions. The modulation of integrin expression and affinity are two such mechanisms in which cells can alter their ability to interact with other cells and ECM. We show in vitro that transmigrated T cells exhibit down-regulation of very late activation antigen-4 and leukocyte function-associated antigen-1 integrin surface expression and a decrease in binding to recombinant vascular cell adhesion molecule-1 and recombinant intercellular adhesion molecule-1. Also, transmigrated T cells displayed an increase in binding to collagens I and IV and fibronectin. Further, brain sections of experimental autoimmune encephalomyelitis mice demonstrated that as T cells migrated farther into the tissue, very late activation antigen-4 expression was lost while CD4 expression remained unchanged. The significance of these findings in the modulation of the inflammatory response is discussed.

Asunto(s)

Adhesión Celular , Endotelio Vascular/fisiología , Matriz Extracelular/fisiología , Linfocitos T/fisiología , Animales , Encéfalo/inmunología , Encéfalo/patología , Movimiento Celular , Células Cultivadas , Colágeno/fisiología , Encefalomielitis Autoinmune Experimental/inmunología , Encefalomielitis Autoinmune Experimental/patología , Endotelio Vascular/inmunología , Fibronectinas/fisiología , Citometría de Flujo , Integrina alfa4beta1 , Integrinas/biosíntesis , Molécula 1 de Adhesión Intercelular/farmacología , Interleucina-2/farmacología , Antígeno-1 Asociado a Función de Linfocito/biosíntesis , Ratones , Ratones Endogámicos , Proteína Básica de Mielina/farmacología , Fragmentos de Péptidos/farmacología , Receptores Mensajeros de Linfocitos/biosíntesis , Receptores de Antígeno muy Tardío/biosíntesis , Proteínas Recombinantes/farmacología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Regulación hacia Arriba , Molécula 1 de Adhesión Celular Vascular/farmacología , Molécula 1 de Adhesión Celular Vascular/fisiología

RESUMEN

T cells play an important role in the pathogenesis of diabetes in the nonobese diabetic (NOD) mouse. CD8 cytotoxic T cell lines and clones were generated from the lymphocytic infiltrate in the islets of Langerhans of young (7-wk-old). NOD mice by growing them on (NOD x B6-RIP-B7-1)F1 islets. These cells proliferate specifically to NOD islets and kill NOD islets in vitro. The cells are restricted by H-2Kd, and all bear T cell antigen receptor encoded by V beta 6. When these CD8 T cell lines and clones are adoptively transferred to irradiated female NOD, young NOD-SCID, and CB17-SCID mice, diabetes occurs very rapidly, within 10 d of transfer and without CD4 T cells.

Asunto(s)

Antígeno B7-1/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Diabetes Mellitus Tipo 2/inmunología , Islotes Pancreáticos/inmunología , Secuencia de Aminoácidos , Animales , Antígeno B7-1/genética , Secuencia de Bases , Células Clonales , Citocinas/biosíntesis , Femenino , Inmunohistoquímica , Inmunoterapia Adoptiva , Insulina/genética , Activación de Linfocitos , Glicoproteínas de Membrana/biosíntesis , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos NOD , Ratones SCID , Datos de Secuencia Molecular , Páncreas/anatomía & histología , Perforina , Proteínas Citotóxicas Formadoras de Poros , Regiones Promotoras Genéticas/genética

RESUMEN

B7-1 is a co-stimulatory molecule that signals T-cells that recognize antigen to proliferate and differentiate into effector T-cells. The same cell must present antigen and express co-stimulatory molecules, such as B7-1, to activate naive T-cells. Thus, tissues that do not express co-stimulatory molecules would not be expected to induce immune responses, while expression of a co-stimulator on tissue cells may convert them into effective antigen-presenting cells and induce autoimmunity. To test this, transgenic mice have been generated that express B7-1 on the beta-cells of the pancreatic islets of Langerhans. On a B6 genetic background, B7-1 expression on beta-cells does not predispose to diabetes. B6 mice are resistant to diabetes. However, when B7-1 is expressed on the beta-cells of B6 mice backcrossed once to the genetically susceptible NOD strain, the onset of diabetes is accelerated and the autoimmune attack intensified. This illustrates that B7-1 is a very potent co-stimulatory molecule in vivo and that its presence on the surface of tissue cells can potentiate the autoimmune process.

Asunto(s)

Antígeno B7-1/biosíntesis , Diabetes Mellitus Tipo 1/fisiopatología , Islotes Pancreáticos/metabolismo , Linfocitos T/inmunología , Envejecimiento/fisiología , Animales , Cruzamientos Genéticos , Diabetes Mellitus Tipo 1/patología , Femenino , Expresión Génica , Humanos , Islotes Pancreáticos/crecimiento & desarrollo , Islotes Pancreáticos/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos NOD , Ratones Transgénicos , Páncreas/crecimiento & desarrollo , Páncreas/patología

RESUMEN

An adoptive transfer model of insulin-dependent diabetes mellitus (IDDM) in the nonobese diabetic mouse was used to examine the roles of alpha 4-integrin, vascular cell adhesion molecule 1 (VCAM-1); and intercellular adhesion molecule 1 (ICAM-1) in the pathogenesis of autoimmune diabetes. Antibodies specific for both alpha 4-integrin and one of its ligands, VCAM-1, were able to delay onset of diabetes and decrease the incidence of the disease in adoptive transfer studies. This blocking of disease was accompanied by a marked decrease in lymphocytic infiltration of the islets of Langerhans. Furthermore, these antibodies preferentially block entrance of CD4 T cells into the tissue. Antibodies specific for ICAM-1 had little effect on the onset or incidence of IDDM. Thus, we conclude that an alpha 4-integrin-VCAM-1 interaction is important in T cell entry into the islets of Langerhans and in the pathogenesis of IDDM. In addition, the cascade of events leading to T cell transit across endothelium may be different for CD4 and CD8 cells, and may differ depending on the endothelium involved. Our results support the more general conclusion that an alpha 4-integrin-VCAM-1 interaction may be crucial in allowing activated effector CD4T cells to leave the blood and enter tissue to clear infection.

Asunto(s)

Moléculas de Adhesión Celular/fisiología , Diabetes Mellitus Tipo 1/etiología , Integrinas/fisiología , Linfocitos T/fisiología , Animales , Anticuerpos Monoclonales/inmunología , Relación CD4-CD8 , Moléculas de Adhesión Celular/análisis , Inmunoterapia Adoptiva , Integrinas/análisis , Molécula 1 de Adhesión Intercelular , Islotes Pancreáticos/patología , Ratones , Ratones Endogámicos NOD , Receptores de Antígeno muy Tardío/fisiología , Bazo/inmunología , Linfocitos T/química , Linfocitos T/patología , Molécula 1 de Adhesión Celular Vascular

RESUMEN

Tissue cells obtained from the follicular fluids of women undergoing in vitro fertilization and embryo transfer were found to contain interleukin-1 beta mRNA by Northern analysis. Since macrophages are known to produce interleukin-1 beta, we examined the follicular fluids of 20 women undergoing in vitro fertilization as well as tissue sections of normal human ovary for the presence of macrophages and monocytes. Although granulosa-luteal cells predominate in follicular fluid, we found that resident macrophages and monocytes comprise 5-15% of human follicular tissue cells. In addition, we observed that macrophages are present in the human ovarian follicle as well as in the corpus luteum.

Asunto(s)

Líquido Folicular/citología , Interleucina-1/análisis , Macrófagos/química , Monocitos/química , ARN Mensajero/análisis , Northern Blotting , Transferencia de Embrión , Femenino , Fertilización In Vitro , Citometría de Flujo , Humanos , Técnicas para Inmunoenzimas , Interleucina-1/genética , Fase Luteínica , Ciclo Menstrual , Ovario/citología

RESUMEN

Failure to respond to human menopausal gonadotropin (hMG) with adequate ovarian stimulation is associated with a poor prognosis in subsequent cycles in women participating in an in vitro fertilization/embryo transfer program. Sera from 26 menstruating women (mean age 38 +/- 4.3 years) identified as "low responders" with either tubal or male factor infertility, mean baseline FSH values of 11 mIU/mL, and peak serum estradiol levels lower than 300 pg/mL were assessed for specific antibodies to human ovary and gonadotropins. Twenty-five infertile women with tubal or male factor infertility with a good response to hMG served as controls. Ninety-two percent of low responders had antibodies to FSH and 65% had antibodies to LH when assessed by enzyme-linked immunosorbent assay. Similarly, 77% of low responders had ovarian antibodies. No hepatic antibodies were found in the sera of low responders, indicating that the positivity was not a general interaction with cell components. None of the "good responders" had antibodies to gonadotropins or to ovarian or liver tissue. The significant differences in antibodies between the groups supports a possible immunologic cause for low ovarian stimulation response to gonadotropin.

Asunto(s)

Autoanticuerpos/análisis , Hormona Folículo Estimulante/inmunología , Infertilidad Femenina/inmunología , Hormona Luteinizante/inmunología , Menotropinas/farmacología , Ovario/efectos de los fármacos , Adulto , Ensayo de Inmunoadsorción Enzimática , Estradiol/sangre , Femenino , Fertilización In Vitro , Hormona Folículo Estimulante/sangre , Humanos , Inmunohistoquímica , Infertilidad Femenina/sangre , Infertilidad Femenina/fisiopatología , Hormona Luteinizante/sangre , Persona de Mediana Edad , Ovario/inmunología , Ovario/fisiopatología

RESUMEN

An enzyme-linked immunosorbent assay (ELISA) was used to detect ovarian and oocyte antibodies in serum from 45 patients with premature ovarian failure (POF). Control sera were obtained from a similar group of normally cycling women without POF. A specific antibody reaction was found when POF sera were tested against human ovary (47%) or oocytes (47%). A combined total of 69% of the sera were positive for either ovary or oocytes. Fewer sera were positive for antibodies against human thyroid (18%) or human placenta (22%), and virtually no reaction with human liver (4%) was seen. LH antibodies were detected by ELISA against LH in only 3 POF sera that also contained ovarian antibodies. Therefore, gonadotropin antibodies alone do not appear to account for POF. In addition, 2 patients were treated by immunosuppression and became pregnant coincident with a decline in the serum concentration of ovarian antibodies. In summary, the results of this study are consistent with previous immunohistochemical data which indicate that ovarian and oocyte antibodies are common in patients with POF. This supports the concept that some forms of POF are associated with an autoimmune process. Furthermore, detection of ovarian and oocyte antibodies by ELISA may permit routine diagnosis of autoimmune POF and provide a basis for therapy.

Asunto(s)

Anticuerpos/aislamiento & purificación , Enfermedades del Ovario/inmunología , Ovario/inmunología , Adulto , Ensayo de Inmunoadsorción Enzimática , Femenino , Hormona Folículo Estimulante/inmunología , Humanos , Inmunohistoquímica , Infertilidad Femenina/tratamiento farmacológico , Hormona Luteinizante/inmunología , Metilprednisolona/uso terapéutico , Persona de Mediana Edad , Enfermedades del Ovario/complicaciones , Enfermedades del Ovario/terapia , Ovario/fisiopatología , Óvulo/inmunología , Placenta/inmunología , Glándula Tiroides/inmunología

RESUMEN

To identify luteinizing hormone (LH) receptors, a monoclonal antibody (MAb) was produced by immunization of Balb/c mice with rat luteal cell membranes. Hybridomas, produced by a method for proteins of low antigenicity, were selected by competition with [125I]-hCG (LH) for luteal membrane binding. Conditions for analysis of LH receptor antibody (IgG2b isotype) binding by immunohistochemistry with an avidin-biotin-peroxidase complex were examined and results compared to localization of bound hCG, to detect receptors. By light microscopy, both bound hCG and the LH receptor antibody were located on luteal cell surfaces. In addition, the LH receptor antibody was associated with luteal cell cytoplasm. Cell surface membrane binding, but not cytoplasmic staining, was reduced in ovaries from rats injected with hCG. By electron microscopy, LH receptor antibody was observed in patches on luteal cell surface membranes and was associated with polysomes, small vesicles, and occasionally with discrete areas of endoplasmic reticulum. Therefore, detection of LH receptors with bound hCG may be limited to receptors found on cell surfaces, while additional LH receptors are revealed by use of a receptor antibody. The cytoplasmic LH receptor may represent stages in the processing of receptor protein. Furthermore, the methodology used in this study should be generally useful for immunohistochemistry with other MAb to receptors.

Asunto(s)

Anticuerpos Monoclonales/análisis , Hormona Luteinizante/metabolismo , Ovario/ultraestructura , Receptores de HL/metabolismo , Animales , Anticuerpos Monoclonales/inmunología , Femenino , Inmunohistoquímica , Hormona Luteinizante/inmunología , Microscopía Electrónica , Ovario/citología , Ovario/metabolismo , Ratas , Receptores de HL/inmunología

RESUMEN

In the hamster flank organ, the growth of hair and growth of sebaceous glands are androgen-dependent functions. Although dihydrotestosterone (DHT) is known to be a potent stimulator of flank organ growth, there is no information about localization of DHT receptor sites in this organ. The purpose of this study was to use steroid autoradiography to localize DHT receptors in the hamster flank organ. Because steroid hormones are functional when translocated to nuclear receptors, nuclear localization by autoradiography defines receptor sites. In order to be able to visualize autoradiographic grains from radiolabeled androgens around hair follicles, albino hamsters were studied to avoid confusion between the grains and pigment granules which are abundant in the more common Golden Syrian hamster. Mature male hamsters castrated 24 hours earlier were given tritium-labeled dihydrotestosterone ( [3H]DHT). Using the technique of thaw-mount steroid autoradiography, 4-micron unfixed frozen sections were mounted in the dark onto emulsion-coated glass slides and allowed to develop for 4-6 months. [3H]DHT was found to be concentrated over sebocyte nuclei. The label was present peripherally as well as in differentiating sebocytes. There was no nuclear localization of [3H]DHT in animals pretreated with excessive quantities of unlabeled DHT. Steroid metabolites of [3H] DHT were assessed by thin-layer chromatography in paired tissue samples. Most of the label remained with DHT. Uptake was inhibited in the flank organ of hamsters pretreated with unlabeled DHT. Specific DHT receptors in the albino hamster flank organ are located in peripheral and differentiating sebocytes. Steroid autoradiography is a useful tool to study androgen interaction in the skin.

Asunto(s)

Cricetinae , Dihidrotestosterona/análisis , Receptores Androgénicos/análisis , Receptores de Esteroides/análisis , Animales , Autorradiografía , Cromatografía en Capa Delgada , Masculino , Mesocricetus , Glándulas Sebáceas/análisis , Tritio