RESUMEN
Fetal and neonatal alloimmune thrombocytopenia due to mothers' anti-HPA-5b alloimmunization has generally a milder clinical presentation compared to anti-HPA-1a alloimmunization. Nevertheless, a case with infant's death probably due to intracranial haemorrhage has been reported. However, if platelet-specific alloimmunized mothers with prior fetal or neonate injury receive intravenous immunoglobulins during pregnancy, thrombocytopenia in heterozygous fetus and neonate may be prevented. Here are reported 2 cases of anti-HPA-5b fetal-maternal alloimmunization, one with prior fetal death, the other with prior severe fetal intracranial haemorrhage, which were successfully treated with intraveinous immunoglobulins alone during a second pregnancy with HPA-5b incompatibility.
Asunto(s)
Antígenos de Plaqueta Humana , Plaquetas/inmunología , Enfermedades Fetales/terapia , Inmunoglobulinas Intravenosas/uso terapéutico , Enfermedades del Recién Nacido/terapia , Adulto , Femenino , Enfermedades Fetales/inmunología , Humanos , Recién Nacido , Enfermedades del Recién Nacido/inmunología , Masculino , Intercambio Materno-Fetal/inmunología , Embarazo , Trombocitopenia/inmunología , Trombocitopenia/terapiaRESUMEN
Glucocorticoids bind to their receptors and trigger the transcriptional activation or repression of target genes by binding to DNA sequences, the glucocorticoid responsive element (GRE). The murine interferon-beta (Mu-IFN beta) gene in L929 cells can be induced by dexamethasone to give both transcription and translation products specific to murine IFN beta. The 3'-noncoding region of the Mu-IFN beta gene was found to contain a GRE very similar to the consensus GRE sequence involved in glucocorticoid-regulated genes. Gel retardation assays showed that the oligonucleotide corresponding to that GRE competed with the MMTV GRE oligonucleotide for glucocorticoid receptor binding and was supershifted by human antiglucocorticoid receptor antibodies. Transiently transfected murine cells (L929) with the GRE-IFN beta 3' sequence inserted upstream of the thymidine kinase promoter and the chloramphenicol acetyl transferase gene treated with dexamethasone with or without the antiglucocorticoid RU486 and their chloramphenicol acetyl transferase activity assayed, show that this GRE is efficient. We conclude that the Mu-IFN beta gene in L929 murine cells can be induced by dexamethasone, and that the hormone effect may be mediated by the 3'-GRE sequence.
Asunto(s)
Dexametasona/farmacología , Regulación de la Expresión Génica , Interferón beta/genética , Animales , Secuencia de Bases , Línea Celular , Dexametasona/antagonistas & inhibidores , Dexametasona/metabolismo , Interferón beta/metabolismo , Ratones , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ARN Mensajero/análisis , Receptores de Glucocorticoides/metabolismo , Secuencias Reguladoras de Ácidos Nucleicos , TransfecciónRESUMEN
We have reported previously that Moloney virus-transformed cells, when treated for over 200 passages in the presence of low concentrations of mouse interferon-alpha/beta, can be reverted to a stable nonmalignant status. The cells recover full contact inhibition and are unable to raise tumors when grafted in nude mice. In the present report, we show that whether reverted or malignant, these cells contain deleted v-mos oncogenes, which have lost 392 nucleotides. The truncated oncogenes contain a reduced and modified open reading frame but are able, however, to induce tumors when transfected in mouse 3T3 cells. As there is no difference either in the location or in the structure of this modified v-mos, whether yielded by reverted or malignant cells, we postulate that both cell lines derive from the same population and this modification does not play any role in the reversion process obtained through prolonged IFN-dependent selection. We suggest that reversion could be an epigenetic phenomenon, involving the constitutive synthesis of IFN-beta only in the reverted and not in the malignant cells. The continued persistence of such noncancerous cells could result at least partly from a balance involving the expression of v-mos, IFN-beta, and an IFN antagonist, sarcolectin. These reverted cells can undergo an unlimited number of passages, but they must be trypsinized before day 5 in confluent cultures. Thereafter, the cells stop dividing, cannot proliferate anymore, progressively show signs of apoptosis, and die.
Asunto(s)
Transformación Celular Viral/genética , Inhibición de Contacto/efectos de los fármacos , Genes mos , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Ensayo de Inmunoadsorción Enzimática , Fibroblastos/efectos de los fármacos , Interferón beta/antagonistas & inhibidores , Interferón beta/genética , Lectinas/análisis , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Datos de Secuencia Molecular , Transfección , Células Tumorales CultivadasRESUMEN
Starting from the structure of the novel nonpeptide AT1 receptor antagonist DuP 753 (losartan), a new series of potent antagonists was designed. In these compounds the central imidazole nucleus was replaced by the dihydroimidazol-4-one structure. The most active compounds had a spirocyclopentane or a spirocyclohexane ring in position 5. Like the imidazole series, the best substituents were the linear butyl chain in position 1 and the [2'-(tetrazol-5-yl)biphenylyl]methyl group in position 3. Antagonistic activity was assessed by the ability of the compounds to competitively inhibit [125I]AII binding to the AT1 subtype receptor and to antagonize AII-induced contractions in rabbit aorta rings. The most active compounds had IC50 values in the nanomolar range. In conscious rats, compounds 4 and 21 antagonized the AII pressor response when administered orally. Compound 21 (SR 47436) was the most active; it was recently shown to also be active in cynomolgus monkeys both intravenously and orally. This molecule is now undergoing clinical trials for the treatment of hypertension.
Asunto(s)
Angiotensina II/metabolismo , Antagonistas de Receptores de Angiotensina , Imidazoles/síntesis química , Imidazoles/farmacología , Animales , Compuestos de Bifenilo/síntesis química , Compuestos de Bifenilo/farmacología , Femenino , Técnicas In Vitro , Irbesartán , Macaca fascicularis , Masculino , Conejos , Ratas , Ratas Sprague-Dawley , Receptores de Angiotensina/metabolismo , Sensibilidad y Especificidad , Relación Estructura-Actividad , Tetrazoles/síntesis química , Tetrazoles/farmacologíaRESUMEN
The aim of this pilot study was to assess the feasibility of a tolerance study of qualified (secured by quarantine or solvent-detergent-treated) fresh frozen plasma (FFP) in real conditions of use. We included all patients receiving qualified FFP during a one-month observation period in three french hospitals (Besançon, Brest, Lyon). The 192 FFP transfusion episodes corresponded to 111 patients. Only two thirds of all prescriptions corresponded to indications mentioned in the ministerial order of december 1991. Forty-two episodes consisted of FFP only; in the 150 remaining episodes, at least one product (mostly labile blood products) had been injected within 24 hours before or after the plasma injection. The free interval between FFP transfusion and the nearest associated product was usually less than three hours. Only one side effect was notified. This pilot study points out the difficulties of a tolerance study of qualified FFP in real conditions of use. It also raises the necessity to clarify current indications of FFP.
Asunto(s)
Plasma , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Estudios de Factibilidad , Francia , Humanos , Lactante , Persona de Mediana Edad , Proyectos PilotoRESUMEN
In France, relative increases in hemolytic disease of the newborn (HDN) due to non anti-D maternal alloimmunization is linked to a systematic search and prevention of an anti-D alloimmunization in Rh negative women during and after pregnancy. In May 1985, detection of anti-red blood cell alloantibodies in all pregnant women has been ordered by a new law. We have observed the efficiency of this law by studying the number of HDN with anti-D and non anti-D maternal alloimmunization detected before and after delivery during the years 1984 and 1986. Our results show that this law is well used. Nevertheless, this work demonstrates that often clinicians omit to set cord blood aside for hemoglobin and bilirubinemia concentration determinations. Because these two parameters are very important in establishing the severity of HDN and therapeutic management, improvements are necessary.
Asunto(s)
Eritroblastosis Fetal/prevención & control , Isoanticuerpos/análisis , Embarazo/inmunología , Isoinmunización Rh , Adulto , Femenino , Humanos , Recién NacidoRESUMEN
Thymidine kinase deficient L cells treated by interferon become resistant to transformation by a recombinant plasmid carrying the herpes virus tk gene. The number of colonies appearing after transformation is inversely proportional to the interferon concentration.
Asunto(s)
Interferones/farmacología , Plásmidos/efectos de los fármacos , Timidina Quinasa/genética , Transformación Genética/efectos de los fármacos , Animales , Herpesviridae/genética , Células L , Ratones , Recombinación GenéticaRESUMEN
Specific genes can be introduced into cultured mammalian cells by DNA transfer, sometimes to be stably integrated and expressed. Because interferon inhibits the transformation initiated by DNA viruses, we wondered if it might also affect transformation induced by recombinant plasmids and cellular genes. We show here that in mouse L cells interferon treatment prevents stable integration and expression of the transfected plasmids containing the cloned herpesvirus thymidine kinase (tk) gene, or the dihydrofolate reductase (dhfr) gene from hamster ovary cells. In contrast, interferon does not prevent transient expression of the tk gene in unintegrated form.
Asunto(s)
Transformación Celular Viral/efectos de los fármacos , Genes Virales/efectos de los fármacos , Interferones/farmacología , Animales , Cinética , Células L/efectos de los fármacos , Células L/fisiología , Ratones , Plásmidos , Timidina Quinasa/genética , TransfecciónRESUMEN
A female baby with a severe thrombocytopenia at 18 x 10(9)/l was born to a 29-year-old (gestation 2/partum 2) mother. Scattered petechiae were present on her legs, arms, chest and face, but there was no bleeding, infection, fever or hepatosplenomegaly. A platelet antibody screening immunocapture test was positive, which was performed on the mother's serum 3, 12 and 38 days after delivery, but no platelet-specific antibodies were found by the monoclonal-antibody-specific immobilization of platelet antigen assay. The baby's platelets and lymphocytes and the father's platelets reacted strongly with the HLA antibodies present in the mother's serum. The neonate was treated with intravenous human immunoglobulin (Tegeline), 1 g/kg per day) 1, 2 and 3 days after delivery. The platelet count rose from 18 x 10(9)/l on day 0 to 37 x 10(9)/l on day 3 and to 227 x 10(9)/l on day 12. No platelet transfusion was needed. Several factors which developed hereafter lead us to think that this neonatal alloimmune thrombocytopenia is due to the transplacental passage of maternal HLA antibodies to the baby.
Asunto(s)
Antígenos HLA/inmunología , Isoanticuerpos/sangre , Trombocitopenia/inmunología , Adulto , Plaquetas/inmunología , Femenino , Antígenos HLA/efectos adversos , Humanos , Inmunoglobulinas Intravenosas/administración & dosificación , Recién Nacido , Masculino , Intercambio Materno-Fetal/inmunología , Padres , Embarazo , Sistema del Grupo Sanguíneo Rh-Hr/inmunología , Trombocitopenia/tratamiento farmacológico , Trombocitopenia/etiología , Resultado del TratamientoRESUMEN
During the last thirty years, the diagnosis, management and prevention of haemolytic disease of the newborn infant (HDN) have improved. From 1959 to 1988, 3004 HDN (ABO excluded) have been collected. The percentage of HDN with anti-D alloimmunization decreased significantly (98.4% from 1959 to 1968, 93.5% from 1969 to 1978 and 68.1% from 1979 to 1988). The anti-D HDN with exchange transfusion (ET) fell significantly between the first and second periods (577 versus 970; chi 2 = 19.92; P less than 0.001). On the other hand, the number of HDN other than anti-D increased during these three periods, but the percentage of these HDN which needed ET decreased. Our study shows the long term efficiency of the prevention of anti-D alloimmunization (since 1970) and of the irregular antibodies screening among all pregnant women (since 1979).
Asunto(s)
Eritroblastosis Fetal/prevención & control , Isoinmunización Rh/prevención & control , Anticuerpos Antiidiotipos/uso terapéutico , Especificidad de Anticuerpos , Transfusión de Sangre Intrauterina , Eritroblastosis Fetal/epidemiología , Eritroblastosis Fetal/inmunología , Eritroblastosis Fetal/terapia , Femenino , Humanos , Inmunoglobulina G , Recién Nacido , Isoanticuerpos/inmunología , Embarazo , Diagnóstico Prenatal , Isoinmunización Rh/tratamiento farmacológicoRESUMEN
Hemolytic disease of the newborn infant (HDN) due to anti-K (Cellano) is very uncommon in Caucasians. We report here a case of anti-K HDN. The anti-K alloimmunization appeared in the mother during her fifth pregnancy. This HDN needed an exchange transfusion immediately after delivery. The clinical outcome of the newborn infant was good.
Asunto(s)
Incompatibilidad de Grupos Sanguíneos/inmunología , Eritroblastosis Fetal/inmunología , Isoantígenos , Sistema del Grupo Sanguíneo de Kell , Adulto , Eritroblastosis Fetal/terapia , Recambio Total de Sangre , Femenino , Humanos , Recién Nacido , MasculinoRESUMEN
Interferon sensitivity of different normal and aneusomic human cells and of different mouse-human hybrids cells has been compared. G21 trisomic cells are more sensitive than diploid cells; whereas, on the contrary, triploid cells are normal in their human interferon sensitivity. Among other aneusomic cell lines tested, E16 trisomic cells are significantly less sensitive. These data are in favor of the hypotheses that the G21 chromosome carries genetic information for structural proteins involved in the receptor system for interferon, that there is a regulatory mechanism governing the antiviral state, and that the E16 chromosome is a possible candidate for carrying information for such a depressive regulatory mechanism. None of the chromosome abnormalities studies are involved with interferon synthesis.
Asunto(s)
Cromosomas/metabolismo , Genes , Interferones/metabolismo , Línea Celular , Diploidia , Relación Dosis-Respuesta a Droga , Células Híbridas/efectos de los fármacos , Células Híbridas/metabolismo , Interferones/farmacología , Cariotipificación , Trisomía , Interferencia ViralRESUMEN
Analysis of a great number of independent hamster-human and mouse-chimpanzee somatic cell hybrid clones confirms the role of chromosome 9 as carrying one or more primate beta interferon genes. The presence of chromosome 13 in producing hybrids and its absence in all non producing clones must be kept in mind for future studies. The strong negative regulation of interferon production in the parental hamster cells also affects the human gene product. The UV irradiation target for these regulatory genes is significantly greater than the structural genes responsible for interferon production.
Asunto(s)
Cromosomas Humanos 13-15/análisis , Cromosomas Humanos 6-12 y X/análisis , Interferones/biosíntesis , Pan troglodytes/genética , Animales , Cricetinae , Humanos , Interferones/genética , RatonesRESUMEN
The karyotype of Cercopithecus aethiops was analysed by RHG, GTG, and C banding, and a band nomenclature is proposed.
Asunto(s)
Cercopithecus , Cariotipificación , Animales , HaplorrinosRESUMEN
Four clones of a somatic monkey-mouse hybrid cell line were studied. One of these clones produced both mouse and monkey interferon, while the three others produced only mouse interferon. All four were, however, sensitive to both mouse and monkey (or human) interferon. The karyotype analysis of these four hybrid clones and the parental cell lines enabled us to locate the possible genetic site governing the synthesis of monkey interferon on a small subtelocentric monkey chromosome. The genetic site responsible for the synthesis of the antiviral protein is located on a different (monkey or mouse) chromosome.
Asunto(s)
Cromosomas , Células Clonales/metabolismo , Animales , Línea Celular/metabolismo , Efecto Citopatogénico Viral/efectos de los fármacosRESUMEN
Haemolytic disease of the newborn (HDN) due to Rhesus anti-e alone is rarely observed in the Caucasian population. We report here a case of an R2R2 mother who had never been transfused and whose two children had a mild HDN without transfusion or blood exchange transfusion (BE) after delivery.
Asunto(s)
Eritroblastosis Fetal/inmunología , Isoinmunización Rh/inmunología , Femenino , Humanos , Recién Nacido , MasculinoRESUMEN
The murine interferon-A11 (Mu IFN-A11) gene is a member of the IFN-A multigenic family. In mouse L929 cells, the weak response of the gene's promoter to viral induction is due to a combination of both a point mutation in the virus responsive element (VRE) and the presence of negatively regulating sequences surrounding the VRE. In the distal part of the promoter, the negatively acting E1E2 sequence was delimited. This sequence displays an inhibitory effect in either orientation or position on the inducibility of a virus-responsive heterologous promoter. It selectively represses VRE-dependent transcription but is not able to reduce the transcriptional activity of a VRE-lacking promoter. In a transient transfection assay, an E1E2-containing DNA competitor was able to derepress the native Mu IFN-A11 promoter. Specific nuclear factors bind to this sequence; thus the binding of trans-regulators participates in the repression of the Mu IFN-A11 gene. The E1E2 sequence contains an IFN regulatory factor (IRF)-binding site. Recombinant IRF2 binds this sequence and anti-IRF2 antibodies supershift a major complex formed with nuclear extracts. The protein composing the complex is 50 kDa in size, indicating the presence of IRF2 or antigenically related proteins in the complex. The Mu IFN-A11 gene is the first example within the murine IFN-A family, in which a distal promoter element has been identified that can negatively modulate the transcriptional response to viral induction.
Asunto(s)
Interferón-alfa/genética , Regiones Promotoras Genéticas , Proteínas Represoras , Factores de Transcripción , Animales , Secuencia de Bases , Sitios de Unión , Línea Celular , Proteínas de Unión al ADN/metabolismo , Factor 2 Regulador del Interferón , Ratones , Datos de Secuencia Molecular , Proteínas Nucleares/metabolismoRESUMEN
Comparative transfection analysis of murine interferon A4 and interferon A11 promoter constructs transiently transfected in mouse L929 and human HeLa S3 cells infected with Newcastle disease virus showed that the second positive regulatory domain I-like domain (D motif), located between nucleotides -57 and -46 upstream of the transcription start site, contributes to the activation of virus-induced transcription of the interferon (IFN)-A4 gene promoter by cooperating with the positive regulatory domain I-like and TG-like domains previously described. Electrophoretic mobility shift assay performed with the virus-inducible fragments containing these motifs indicated that the binding activity that we have denoted as virus-induced factor (Génin, P., Bragança, J., Darracq, N., Doly, J., and Civas, A. (1995) Nucleic Acids Res. 23, 5055-5063) is different from interferon-stimulated gene factor 3. It binds to the D motif but not to the virus-unresponsive form of the D motif disrupted by a G-57 --> C substitution. We show that the low levels of IFN-A11 gene expression are caused essentially by the lack of two inducible enhancer domains disrupted by the A-78 --> G and the G-57 --> C substitutions. These data suggest a model taking account of the differential regulation of IFN-A gene family members. They also suggest that virus-induced factor may correspond to the primary transcription factor directly activated by virus that is involved in the initiation of IFN-A gene transcription.
Asunto(s)
Proteínas de Unión al ADN/fisiología , Regulación Viral de la Expresión Génica , Interferón-alfa/genética , Virus de la Enfermedad de Newcastle/fisiología , Regiones Promotoras Genéticas , Factores de Transcripción , Proteínas Virales/fisiología , Animales , Secuencia de Bases , Sitios de Unión/genética , Proteínas de Unión al ADN/metabolismo , Dimerización , Células HeLa , Humanos , Factor 2 Regulador del Interferón , Ratones , Datos de Secuencia Molecular , Proteínas Represoras/fisiología , TATA Box , Transcripción Genética , TransfecciónRESUMEN
Dose-response curves of interferons in different sensitive cells are regularly sigmoidal. In somatic monkey-mouse hybrid cells, however, a significant decrease in the slope of the curve for primate interferon was observed, while the dose-response effect was unaltered for mouse interferon. High concentrations of primate interferon were 10- to 100-times less effective in hybrid clones than in parental monkey CV-1 cells; at low concentrations the antiviral effect was 10- to 20-times higher in hybrid clones than in the parental cells. The receptor(s) for primate interferon located on the cell membrane was destroyed by trypsin but not by EDTA. Similarly, acid pH inactivated these receptor sites. We, thus, postulate that the antiviral effect is, at least partially, related to the amount of interferon taken up by the cells. Uptake could be conditioned by active cooperation of two cell-specific factors: a receptor and an activator. The activator might be missing or inactivated for primate interferon in the hybrid cells. We suggest that the putative antiviral protein is not cell-species specific, and that information for its synthesis in the hybrid cells might be located on a mouse rather than a monkey chromosome.