RESUMEN
An organ culture technique was used to investigate the migration and the morphological evolution of lymphocytes from lymphopoietic tissues. This evolution was compared with the behavior of cells extracted from the tissue and kept in nutritive medium in vitro. It was found that cells were continuously migrating from the fragments of lymph nodes or spleen, and were attaching to the glass. They spread on glass, their protoplasm enlarged and their nucleus became clearer. The evolution towards blastoid cells was identical with that described under artificial stimulation by PHA for example. Cytological identification of the cells actively engaged in antibody synthesis (as detected by local hemolysis in gum) at the time of staining, showed that several distinct cellular types were active, including plasma cells and macrophagelike cells. It is assumed that the stimulated lymphocytes, after spontaneous migration from the tissue are able to evolve into an "immunoblast" stage and then, eventually after fixation upon a physical support, to initiate antibody synthesis.
Asunto(s)
Formación de Anticuerpos , Técnicas de Cultivo , Ganglios Linfáticos , BazoRESUMEN
By combining a tissue culture method with the detection of antibody-producing cells by local hemolysis in gum it has been possible to follow the immunological activity of cells from tissue fragments for long period of time. These fragments were obtained from lymph nodes or spleens of rabbits immunized by sheep erythrocytes. It was found that, while the immunological activity of the free cells in suspensions decreased fast and disappeared in a few days, the cells attaching on glass could express their activity for at least 3 wk. It is assumed that these cells are the daughters of cells from the fragments which were not active antibody producers at the beginning, but differentiated, during the culture, into cells endowed with two capacities: glass adherence and antibody synthesis. One can further admit that the type of culture employed exerts a selective pressure favoring formation of antibody-producing cells.
Asunto(s)
Formación de Anticuerpos , Técnicas de Cultivo , Ganglios Linfáticos , BazoRESUMEN
Neuraminidase (acylneuraminyl hydrolase, EC 3.2.1.18) from the influenza virus A/Hong Kong/68 (H3N2) was purified after treatment of the purified virus with sarcosyl (sodium laurylsarcosinate), centrifugation at 110 000 x g, and chromatography on DEAE-Sephadex and Sephadex G-200. It migrated as a single component during electrophoresis on polyacrylamide gel, and its molecular weight was estimated about 270 000. The enzyme was thermolabile, the activity being reduced to 60% in 10 min at 50 degrees C. The purified neuraminidase had an apparent Km value of 4.1 . 10(-3) M for 5-N-acetyl-2-O-(3-methoxyphenyl)-alpha-D-neuraminic acid and was able to release sialic acid with linkages alpha 2-3, alpha 2-6 and alpha 2-8 (with very different efficiency) from fetuin, gangliosides, colominic acid, and bovine and porcine submaxillary mucins. The enzymic activity was measured by several procedures: (A) spectrophotometric determination at 340 nm of the NADH produced in the reaction catalysed by beta-galactose dehydrogenase on beta-galactose + NAD+, this beta-galactose was the product released from lactose by beta-galactosidase and lactose was the product of the neuraminidase activity on N-acetylneuraminyl-lactose; (B) determination of the colored quinone yielded by the liberated methoxyphenol with 4-aminoantipyrine (Santer, U.V., Yee-Foon, J. and Glick, M.C. (1978) Biochim. Biophys. Acta 523, 435-442); (C) periodate-thiobarbiturate procedures (Warren, L. (1959) J. Biol. Chem 234, 1971-1975 or Aminoff, D. (1961) Biochem. J. 81, 384-391). Some peculiarities of these methods are discussed.
Asunto(s)
Subtipo H3N2 del Virus de la Influenza A , Virus de la Influenza A/enzimología , Neuraminidasa/metabolismo , Gangliósidos , Glicoproteínas , Cinética , Ácidos Neuramínicos , Especificidad por SustratoRESUMEN
Influenza virus type C (Johannesburg/1/66) was used as a source for the enzyme O-acetylesterase (EC 3.1.1.53) with several natural sialoglycoconjugates as substrates. The resulting products were immediately employed as substrates using influenza virus type A [(Singapore/6/86) (H1N1) or Shanghai/11/87 (H3N2)] as a source for sialidase (neuraminidase, EC 3.2.1.18). A significant increase in the percentage of sialic acid released was found when the O-acetyl group was cleaved by O-acetylesterase activity from certain substrates (bovine submandibular gland mucin, rat serum glycoproteins, human saliva glycoproteins, mouse erythrocyte stroma, chick embryonic brain gangliosides and bovine brain gangliosides). A common feature of all these substrates is that they contain N-acetyl-9-O-acetylneuraminic acid residues. By contrast, no significant increase in the release of sialic acid was detected when certain other substrates could not be de-O-acetylated by the action of influenza C esterase, either because they lacked O-acetylsialic acid (human glycophorin A, alpha 1-acid glycoprotein from human serum, fetuin and porcine submandibular gland mucin) or because the 4-O-acetyl group was scarcely cleaved by the viral O-acetylesterase (equine submandibular gland mucin). The biological significance of these facts is discussed, relative to the infective capacity of influenza C virus.
Asunto(s)
Acetilesterasa/metabolismo , Gammainfluenzavirus/enzimología , Virus de la Influenza A/enzimología , Neuraminidasa/metabolismo , Ácidos Siálicos , Especificidad por SustratoRESUMEN
Seventy-eight healthy subjects belonging to 16 different families were submitted to an anti-influenza vaccination. The antibody titers and the red blood cell and plasma Mg concentrations were determined before and 30 days after vaccination. The population study performed on 32 subjects showed the occurrence of a higher antibody response (P less than 0.01) and a lower red blood cell Mg level, among the Bw35 individuals. These findings are confirmed by family studies: HLA identical sibs have values much closer to those of the propositi than to those of the HLA different sibs. The relationships between HLA, immune response, and Mg revealed by the present investigation are discussed in light of the literature together with the known associations between HLA Bw35 antigen and diseases.
Asunto(s)
Anticuerpos Antivirales/inmunología , Antígenos HLA/inmunología , Magnesio/sangre , Adulto , Niño , Eritrocitos/análisis , Femenino , Antígeno HLA-B35 , Humanos , Inmunización , Masculino , Orthomyxoviridae/inmunología , Factores Sexuales , Estadística como AsuntoRESUMEN
BACKGROUND: The Regional Influenza Surveillance Group (GROG) is a French surveillance network set up in 1984. It collects virological specimens and health service based indicators on a weekly basis. Our aim was to assess the predictive value of the health service based indicators for the detection of influenza A epidemics. METHODS: Virological data were used as a gold standard for defining the epidemics. For each health service based indicator, a statistical threshold was used as a test for the identification of epidemic weeks. Finally, an epidemiological criterion was defined in order to improve the specificity and the speed of detection of outbreaks. RESULTS: Health service based indicators have a positive predictive value of around 0.80. They also advance the detection of outbreaks by between 1 and 4 weeks. CONCLUSIONS: These indicators are easy to collect and are useful for the surveillance of influenza epidemics. Such a system is the prerequisite for the rational use of preventive tools.
Asunto(s)
Brotes de Enfermedades , Indicadores de Salud , Virus de la Influenza A , Gripe Humana/epidemiología , Vigilancia de la Población/métodos , Estudios de Evaluación como Asunto , Humanos , Gripe Humana/prevención & control , Gripe Humana/virología , Paris/epidemiología , Probabilidad , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
In previous serological surveys of lemurs in Madagascar, antibodies against flaviviruses were frequently detected. To examine the epidemiological role of Lemur fulvus, experimental infections with yellow fever (YF) virus and West Nile (WN) virus were performed. YF and WN infections were clinically unapparent. A 3 to 4-day-long viremia, with moderate levels was observed with YF virus. WN virus, especially the strain isolated in Madagascar, provoked a 4 to 6-day-long viremia sufficient to infect Aedes aegypti. In all experiments, the antibody response was studied during the following weeks by 3 methods. The results led to the conclusion that Malagasy lemurs could act as amplifying hosts for WN virus present in Madagascar, and as hosts for YF virus if it were introduced on the island. The epidemiological role of these primates is discussed according to their ecology and their contact with potential mosquito vectors in forest areas of Madagascar.
Asunto(s)
Lemur/microbiología , Lemuridae/microbiología , Infecciones por Togaviridae/microbiología , Fiebre del Nilo Occidental/microbiología , Fiebre Amarilla/microbiología , Aedes/microbiología , Animales , Anticuerpos Antivirales/análisis , Culex/microbiología , Culicidae/microbiología , Insectos Vectores/microbiología , Madagascar , Ratones , Fiebre del Nilo Occidental/inmunología , Fiebre del Nilo Occidental/transmisión , Virus del Nilo Occidental/inmunología , Fiebre Amarilla/inmunología , Fiebre Amarilla/transmisión , Virus de la Fiebre Amarilla/inmunologíaRESUMEN
Neuraminidase is one of the two surface glycoproteins of influenza virions. In order to compare neuraminidases of the same subtype but isolated from different species (man, birds, pig), a new and simple method was adapted and optimized using peanut hemagglutinin. Results were very similar to those obtained with the classical method recommended by the WHO, using fetuin as a substrate. The technique was used to examine the relationship between animal and human neuraminidases belonging to serotypes N1 and N2. The results confirm the possible role of ducks as a reservoir for influenza viruses and the eventuality of interspecific exchanges.
Asunto(s)
Neuraminidasa/análisis , Orthomyxoviridae/enzimología , Animales , Variación Antigénica/inmunología , Patos , Humanos , Virus de la Influenza A/clasificación , Virus de la Influenza A/enzimología , Lectinas , Neuraminidasa/inmunología , Orthomyxoviridae/clasificación , Aglutinina de Mani , Especificidad de la Especie , PorcinosRESUMEN
In order to provide an easy and powerful analysis of influenza C viral HEF RNA segment of a recent strain, a combination of reverse transcription and the polymerase chain reaction was used. We amplified the entire coding region of the HEF gene of a laboratory strain of virus called C/Johannesburg/1/66, widely used for binding and esterase activity studies as well as that of a strain isolated in 1991 (C/Paris/145/91) from a patient suffering from severe flu syndrome. The sequences we amplified were about 2 kilobases long. In this work, we show that the forward 'universal primer' Uni1, which has been used for influenza A and B viruses cDNA syntheses can also be used for influenza C virus. The PCR primers were designed to contain restriction sites to make the PCR products ready to be used for further purposes. A restriction analysis of the PCR products combined with analyses of all the human influenza C virus HEF gene sequences published so far permitted the design of sets of oligonucleotides which can prime PCR on cDNA of unknown influenza C virus for cloning.
Asunto(s)
Gammainfluenzavirus/genética , Reacción en Cadena de la Polimerasa/métodos , ARN Viral/genética , Virología/métodos , Secuencia de Bases , Enzimas de Restricción del ADN , ADN Viral/genética , Amplificación de Genes , Genes Virales , Humanos , Gripe Humana/microbiología , Gammainfluenzavirus/aislamiento & purificación , Datos de Secuencia MolecularRESUMEN
A new membrane-enzyme immunofiltration assay (MIFA) was developed for rapid diagnosis of influenza A infection. The pretreated specimens were dispensed into a 1.2 micron Biodyne B nylon membrane-bottomed microplate and vacuum filtration was applied. Blocking solution, peroxidase-conjugated anti-influenza A nucleoprotein monoclonal antibody, washing buffer and substrate were added in that order. The assay was completed within 30 min. Out of 103 nasopharyngeal swabs collected in transport medium, 31 isolates of influenza A virus were obtained and 22 specimens were detected directly by the MIFA technique. The 9 isolation-positive MIFA-negative specimens required 6 days or more for viral detection in cell culture, and probably contained a very low quantity of virus. The 72 cell culture negative specimens were also negative by MIFA. Comparison with a classical immunocapture assay (ICA) gave a better sensitivity for MIFA, as only 15/103 specimens were positive by ICA. MIFA is a rapid test with 71% sensitivity and 100% specificity. It was also very useful to test the cell culture supernatants, as a sensitivity of 100% was obtained with MIFA when the immunofluorescence technique was positive. The same technique could be readily carried out on the same plate for other respiratory viruses since capture antibody is not used.
Asunto(s)
Antígenos Virales/análisis , Técnicas para Inmunoenzimas , Virus de la Influenza A/aislamiento & purificación , Gripe Humana/diagnóstico , Nasofaringe/microbiología , Adolescente , Adulto , Niño , Preescolar , Humanos , Lactante , Virus de la Influenza A/inmunología , Persona de Mediana Edad , Nylons , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
The effects of severe and moderate iron deficiency upon the antibody response to influenza virus were investigated in rats. Three groups of weanling male Wistar rats were fed one of two iron-deficient diets (5 mg and 15 mg iron/kg diet) or a normal iron-containing diet (35 mg iron/kg diet). A group of individually pair-fed rats was introduced with the low iron-consuming rats. The effects of the diets upon various iron status parameters were followed during the 4th, 5th, 6th, and 7th week of diet. After 4 weeks of feeding different diets, an intraperitoneal injection of inactivated influenza virus A/New Jersey/76 was performed and a recall injection was done at 5 weeks. Primary and secondary antibody responses were assayed. Rats were sacrificed at 7 weeks of diet. After 4 weeks of feeding different diets, the rats fed the 5 mg iron/kg diet were severely anemic and rats fed 15 mg iron/kg diet were moderately iron-deficient, as shown by their iron status parameters. Growth was delayed in anemic and matched pair-fed rats. A primary antibody response was almost nonexistent in all groups. Secondary antibody titers were significantly weaker in anemic rats than in ad libitum controls, but were not different from those of pair-fed rats. This response was similar in moderately iron-deficient, ad libitum, and pair-fed rats. These results show that antibody synthesis in response to the influenza virus vaccine is preserved in moderate iron deficiency but is reduced in severe anemia. The reduction in energy consumption associated with severe iron deficiency in the rat could play a part in the altered humoral response.
RESUMEN
Sera collected in May 1984 from 132 adult residents of Karamoja district, Uganda, were examined by haemagglutination inhibition tests for antibodies against selected arboviruses, namely Chikungunya and Semliki Forest alphaviruses (Togaviridae); dengue type 2, Wesselsbron, West Nile, yellow fever and Zika flaviviruses (Flaviviridae); Bunyamwera, Ilesha and Tahyna bunyaviruses (Bunyaviridae); and Sicilian sandfly fever phlebovirus (Bunyaviridae); and by immunofluorescence tests against certain haemorrhagic fever viruses, Lassa fever arenavirus (Arenaviridae), Ebola-Sudan, Ebola-Zaïre and Marburg filoviruses (Filoviridae), Crimean-Congo haemorrhagic fever nairovirus and Rift Valley fever phlebovirus (Bunyaviridae). Antibodies against Chikungunya virus were the most prevalent (47%), followed by flavivirus antibodies (16%), which were probably due mainly to West Nile virus. No evidence of yellow fever or dengue virus circulation was observed. A few individuals had antibodies against Crimean-Congo haemorrhagic fever, Lassa, Ebola and Marburg viruses, suggesting that these viruses all circulate in the area.
Asunto(s)
Anticuerpos Antivirales/análisis , Infecciones por Arbovirus/epidemiología , Adulto , Alphavirus/inmunología , Infecciones por Arbovirus/inmunología , Bunyaviridae/inmunología , Virus Chikungunya/inmunología , Femenino , Flavivirus/inmunología , Técnica del Anticuerpo Fluorescente , Pruebas de Inhibición de Hemaglutinación , Fiebres Hemorrágicas Virales/epidemiología , Fiebres Hemorrágicas Virales/inmunología , Humanos , Masculino , Infecciones por Togaviridae/epidemiología , Uganda/epidemiología , Virus Zika/inmunologíaRESUMEN
Little is known of the epidemiology of influenza C virus infections in western Europe and of the exact role of this agent in acute viral respiratory infections. Several tests may be used for detecting antibodies against this agent but the significance of their respective results is not clear. A total of 301 samples of serum was collected from persons aged from 4 months to 88 years living in France in 1988. The samples were tested for the presence of antibodies to influenza C virus by haemagglutination-inhibition (HI) tests and ELISA. The specificity of the results was checked by immunoblotting and by antibody absorption with staphylococcal protein A. Significant HI activity was found in 61% of the 301 samples tested, titres ranging from 20-320; 70% were positive by ELISA with titres ranging from 500 to 32,000. The population tested was divided into four age groups: 0-15 years; 16-30 years; 31-50 years and 51-88 years. The highest rates for positive samples were found in the 16-30 year group (76 and 79% by HI tests and ELISA respectively) as well as significant HI and ELISA geometric mean titres. Positive samples were less common in young children (46 and 50% by HI tests and ELISA respectively) and in the oldest group (44 and 54% respectively). The 31-50 years age group formed an intermediate class. The high prevalence of antibody as well as the significant titres indicate intense circulation of influenza C virus, especially among young adults.
Asunto(s)
Gammainfluenzavirus , Infecciones por Orthomyxoviridae/epidemiología , Adolescente , Adulto , Factores de Edad , Anciano , Anticuerpos Antivirales/sangre , Niño , Ensayo de Inmunoadsorción Enzimática , Francia/epidemiología , Pruebas de Inhibición de Hemaglutinación , Pruebas de Hemaglutinación , Humanos , Inmunoglobulina G/metabolismo , Lactante , Recién Nacido , Gammainfluenzavirus/inmunología , Persona de Mediana Edad , Infecciones por Orthomyxoviridae/inmunología , Sensibilidad y Especificidad , Proteína Estafilocócica A/aislamiento & purificación , Proteína Estafilocócica A/metabolismoRESUMEN
Influenza virus strains of different types for use as an inactivated vaccine have been successfully grown in different cell lines. Increasing titres were obtained with BHK-21/BRS, VERO and MDCK cells. Cultures in stationary flasks, in spinner cultures or in large bioreactor systems were tested and the optimal conditions were studied. MDCK cells grown in serum-free medium before and during the virus production phase were found to yield high titres in the presence of trypsin. Satisfactory results were obtained with egg-adapted strains of human and equine origin as well as with strains just isolated from human patients without any further passages in eggs or cell culture.
Asunto(s)
Virus de la Influenza A/inmunología , Vacunas contra la Influenza , Animales , Línea Celular , Chlorocebus aethiops , Cricetinae , Técnicas de Cultivo/métodos , Perros , Caballos , Humanos , Virus de la Influenza A/crecimiento & desarrollo , Virus de la Influenza A/aislamiento & purificación , Células Vero , Virología/métodos , Replicación ViralRESUMEN
Antigenic variation, New Jersey porcine influenza, Pandemic planning: Berlin 1993, Pandemics, Principles of action, H5N1 avian influenza in Hong Kong.
RESUMEN
Two seroepidemiological surveys carried out so far, one in Japan, the other in France, gave a strong indication that dogs may be naturally infected by influenza C virus, considered to be exclusively human until recently. In this work, 101 serum samples were collected during winter 1989/1990 from dogs in Castilla y León, Spain. Sera were tested for the presence of antibodies to influenza C virus by Hemagglutination Inhibition (HI) test. Using antibody absorption by staphylococcal protein A, we demonstrated the specificity of the results. Significant HI activity was found in 56.3% of the 101 tested sera and titres ranged from 25 to 200.
Asunto(s)
Anticuerpos Antivirales/sangre , Enfermedades de los Perros/epidemiología , Gammainfluenzavirus , Infecciones por Orthomyxoviridae/veterinaria , Animales , Enfermedades de los Perros/microbiología , Perros , Pruebas de Inhibición de Hemaglutinación/veterinaria , Pruebas de Hemaglutinación/veterinaria , Gammainfluenzavirus/inmunología , Infecciones por Orthomyxoviridae/epidemiología , Sensibilidad y Especificidad , Estudios Seroepidemiológicos , España/epidemiologíaRESUMEN
The evaluation of morbidity risk is difficult in influenza because of a lack of accurate quantitative information and of a high variability of the virus. The level of specific herd immunity towards epidemic strains is an important factor of prevision. As far as mortality is concerned, there is a net and regular increase of risk up from the age of 45. In addition to elderly, other high risk categories are known such as patients suffering of a number of organic deficiences or chronical conditions, pregnant women and children. The best evaluation of efficacy of vaccines is the study of incidence reduction in a vaccinated group in comparison to unvaccinated controls. Present vaccines are really efficient to reduce the risk of getting the disease, except in the case of an antigenic shift, which means the activity of a new antigenic component. These advantages are to be weighted to the small risk of vaccine reactions, which are not frequent and usually benign. Several vaccination strategies can be used. In France, it has been decided to recommand vaccine to priority groups: people over 65 and deficient patients. This policy is eqlivalent in many western countries (West Europe, USA); it is not efficient in preventing or delaying epidemics.
Asunto(s)
Gripe Humana/epidemiología , Anciano , Niño , Femenino , Francia , Humanos , Inmunidad , Vacunas contra la Influenza/efectos adversos , Gripe Humana/inmunología , Persona de Mediana Edad , Orthomyxoviridae , Embarazo , RiesgoRESUMEN
The phenomenon of arbovirus transmission by their vectors is based upon the constitution of virus-vector systems, not only at the species level, but also at an infra-specific level. The study of such systems leads to consider some particular aspects of their ecology which can have important epidemiological consequences (selection of viral clones by the vector, transovarial and sexual transmissions). Finely the authors approach the question of evolution of virus-vector systems, which appear as adaptative, functional and temporary associations.
Asunto(s)
Infecciones por Arbovirus/transmisión , Vectores Artrópodos , Dinámica Poblacional , Aedes/microbiología , Arbovirus , Clima , HumanosRESUMEN
I.--After showing that bank voles are parasited only by Ixodes ricinus larvae, the authors attempt to found different factors (demographic, biometric, and sexual) who favor individual parasitism. The authors conclude to absent of anti tick immunity for this rodent specie. II.--The search for anti-central european encephalitis antibodies (I.H.A.) are shown that 2 p. cent animals were immuns. Yearly and monthly chronologies of antibodies apparition are shown, factors favoring the growth of specific Central european encephalitis antibodies are discussed. III.--The Central european encephalitis tick viral infection of bank vole is studied according to the number of viral strains isolated from different viscera. The monthly chronology of this infection is shown.
Asunto(s)
Arbovirus/patogenicidad , Encefalitis Transmitida por Garrapatas/microbiología , Roedores/microbiología , Animales , Anticuerpos Antivirales/análisis , Arbovirus/inmunología , Encefalitis Transmitida por Garrapatas/inmunología , Europa (Continente) , Francia , Humanos , Inmunidad InnataRESUMEN
The collection of data for influenza surveillance is difficult because clinical cases cannot easily be notified by practitioners and because only a few cases are studied by specific biological methods. In different countries, several indices are used such as: morbidity returns, mortality returns, absenteeism returns preferably in combination. In France, the two National Reference Centres in Paris and Lyon coordinate the activities of all diagnostic laboratories where influenza is detected by collecting and redistributing their findings and by identifying the strains they isolate. Furthermore, a collaborative network of physicians and institutions is being organised to provide a complete coverage of the country. I.N.S.E.R.M. and the National Centres have set up during the last two winters a new method of specific surveillance in school children: it combines observation of absenteeism peak in randomly selected classes and etiologic diagnosis by virus isolation. Surveillance of such a group, particularly sensitive to epidemic spread and probably partly responsible for it, was very efficient: early isolations were obtained and the presence of epidemics was detected before overt outbreaks occurred.