http://repub.eur.nl/res/aut/757/ List of Publications en http://repub.eur.nl/static-eur/img/logo.png http://repub.eur.nl http://repub.eur.nl/res/pub/3293/ 1985-01-01T00:00:00Z Hemagglutination inhibiting (HI) monoclonal antibody preparations (MA) were raised against six influenza A (H3N2) strains from the period 1977-1982. Twenty-three hybridomas were selected and titrated in HI assays against these strains and against 18 influenza A (H3N2) viruses isolated in The Netherlands during the seasons 1981-1982 and 1982-1983. Similar HI tests were performed with conventional post-infection ferret antisera and with ferret antisera adsorbed with heterologous strains of influenza A (H3N2) virus. The resulting serological data were subjected to a computerized taxonomic cluster procedure based on the Euclidean distance between viruses. With respect to the degree of separation between clusters the unadsorbed ferret antisera were inferior to the adsorbed antisera whereas the MA were superior to both. Our results demonstrate that computer programs based on numerical taxonomy can be helpful in processing large numbers of serological data and that MA are indispensable in epidemiological and diagnostic influenza studies. http://repub.eur.nl/res/pub/3302/ 1985-01-01T00:00:00Z An in vitro system of poliovirus-specific antibody production by peripheral blood B cells on stimulation by the virus has been developed. Virus-neutralizing antibodies in culture supernatant fluids, or virus-specific antibody-secreting cells (ASC) were detected by microneutralization assay and ELISA-SPOT test, respectively. After booster immunization with polio vaccine, anti-poliovirus-neutralizing ASC were present in circulation. This response was measurable between 5 and 12 days after booster vaccination. At between 12 and 90 days, another subset of B cells was found in peripheral blood that only produced poliovirus-specific neutralizing antibody after in vitro antigenic stimulation. The in vitro virus-induced response required B cells, monocytes, and T4+ (T helper) cells, and was shown to result from de novo protein synthesis. The anti-poliovirus-neutralizing response in vitro could be dissected in a type-specific and intertypic cross-reactive response by using various antigen concentrations for in vitro stimulation. Evidence was obtained by absorption studies for the existence of intertypic cross-reactive neutralization-inducing epitopes. http://repub.eur.nl/res/pub/3283/ 1984-01-01T00:00:00Z Panels of monoclonal antibodies raised against different poliovirus type 1, 2 and 3 strains, were tested in a micro-neutralization test and in a micro-enzyme linked immunosorbent assay against a large number of poliovirus strains. The results were compared with those obtained with the classical system of serodifferentiation using strain specific cross-absorbed antisera. For this purpose a theoretical pattern fitting computer program was developed, in which each strain could be compared with all the other strains of which the serological data had been stored in the memory of the computer. The results obtained with the panels of monoclonal antibodies coincided well with those obtained with the cross-absorbed antisera. Especially for the identification of virus isolates related to the Sabin vaccine strains, these panels of monoclonal antibodies proved to be valuable tools. http://repub.eur.nl/res/pub/3270/ 1983-01-01T00:00:00Z In the present report an in vitro method for obtaining a secondary human antibody response to a dog kidney cell vaccine against rabies virus (DKCV) is described. Cultures of peripheral blood mononuclear cells from normal rabies-immune and nonimmune donors were stimulated in vitro by DKCV. The production of virus-specific antibody in supernatant fluids was monitored by ELISA. Antibody was produced by lymphocytes from rabies-immune individuals, whereas those of nonimmune subjects consistently failed to produce anti-rabies antibodies after in vitro stimulation with DKCV. The generation of the anti-rabies virus antibody response of lymphocytes stimulated with DKCV was shown to be an antigen-dependent, as well as an antigen-specific process. Optimal antigen-specific responses were observed at relatively low concentrations of antigen (10(-1) to 10(-2) micrograms/culture). At increasing concentrations of antigen in culture (greater than 1 microgram/culture), the anti-rabies virus response was suppressed. Antibody produced upon stimulation was capable of neutralizing rabies virus. The response to rabies virus requires T cell help because lymphocytes depleted of SE rosetting cells did not respond to an antigenic stimulus. Studies in which the same individuals were followed over time showed a sequential development of circulating B cell subsets. The system may provide a model for the study of human B cell differentiation in vivo and in vitro and may be valuable for testing the potency of rabies vaccines in vitro. http://repub.eur.nl/res/pub/3271/ 1983-01-01T00:00:00Z A panel of 10 monoclonal antibodies raised to 3 different poliovirus type 1 strains was tested in a micro-enzyme-linked immunosorbent assay and in a micro-neutralization test against 87 poliovirus type 1 strains. The results, evaluated in a newly developed system for intratypic strain characterization, were compared with the results obtained with the classical sero-differentiation system by using a small number of strain-specific, cross-absorbed antisera. The new system not only uses results obtained with strain-specific antibody preparations, but also uses the information obtained with monoclonal antibodies reacting with less unique antigenic determinants. In a theoretical pattern fitting computer program, each virus strain could be compared with all the other strains for which serological data were stored in the memory of the computer. The results obtained with the new system coincided well with those obtained with the classical system: all except one of the strains classified as Sabin-like or intermediate in the classical system scored 'perfect fit' or 'related' with the Sabin 1 vaccine strain in the new system. Likewise, all virus isolates classified as Kuwait-like in the classical system scored 'perfect fit' or 'related' with the Dutch Kuwait-like isolate strain 78-9030.