A Whole Virus Pandemic Influenza H1N1 Vaccine Is Highly Immunogenic and Protective in Active Immunization and Passive Protection Mouse Models

The recent emergence and rapid spread of a novel swine-derived H1N1 influenza virus has resulted in the first influenza pandemic of this century. Monovalent vaccines have undergone preclinical and clinical development prior to initiation of mass immunization campaigns. We have carried out a series of immunogenicity and protection studies following active immunization of mice, which indicate that a whole virus, nonadjuvanted vaccine is immunogenic at low doses and protects against live virus challenge. The immunogenicity in this model was comparable to that of a whole virus H5N1 vaccine, which had previously been demonstrated to induce high levels of seroprotection in clinical studies. The efficacy of the H1N1 pandemic vaccine in protecting against live virus challenge was also seen to be equivalent to that of the H5N1 vaccine. The protective efficacy of the H1N1 vaccine was also confirmed using a severe combined immunodeficient (SCID) mouse model. It was demonstrated that mouse and guinea pig immune sera elicited following active H1N1 vaccination resulted in 100% protection of SCID mice following passive transfer of immune sera and lethal challenge. The immune responses to a whole virus pandemic H1N1 and a split seasonal H1N1 vaccine were also compared in this study. It was demonstrated that the whole virus vaccine induced a balanced Th-1 and Th-2 response in mice, whereas the split vaccine induced mainly a Th-2 response and only minimal levels of Th-1 responses. These data supported the initiation of clinical studies with the same low doses of whole virus vaccine that had previously been demonstrated to be immunogenic in clinical studies with a whole virus H5N1 vaccine.     

Geographic variation in seed dormancy among populations of Echinochloa crus-galli

In 1991–1993, we investigated the incidence of seed dormancy in 25 local populations of barnyard grass, Echinochloa crus-galli (L.) P.Beauv., in the western Czech Republic. The percentage of germination after 4 months afterripening of dry seeds at 25°C varied between 0.0 and 83.6%. Although there were significant annual differences in the percentage of germination at some localities, typical proportions of dormant seeds persisted over 3 years at field sites where the seed bank was not disturbed. One-way ANOVA (using data from 14 cultivated or abandoned fields) revealed that 73.0% of variance in seed dormancy incidence could be attributed to the effect of locality (P<0.001). Incidence of dormancy was not correlated with mother plant stature (dry above-ground biomass, number of tillers, maximal stem height) nor seed mass. There was a significant correlation (r ²=0.403, P<0.005) between dormancy incidence at natural localities in 1991 and in F₁ offspring sown at experimental grounds at Praha-Ruzyn in 1992. The results indicate that heredity is important in maintaining local variation in seed dormancy, probably favoured by the self-pollinating reproduction of barnyard grass.     

Methodische Untersuchungen zur Ploidiebestimmung an Ruhekernen

Zusammenfassung BeiTrifolium pratense (2n=2x=14) undhybridum (2n=2x=16) ist in jedem haploiden Chromosomensatz ein bestimmtes Chromosom vorhanden, von dem ein bestimmter Abschnitt im ruhenden Zellkern als T-Chromozentrum gut sichtbar sein kann. Die Anzahl dieser T-Chromozentren schwankt innerhalb eines Organs und Gewebes einer Pflanze. Bei einer jeweils bestimmten Prozentzahl der Ruhekerne, die aus gewissen, nachfolgend aufgeführten Geweben und Organen stammen, ist die T-Chromozentrenanzahl gleich der Genomanzahl oder Ploidiestufe der betreffenden Einzelpflanze. Höher als die Genomanzahl wird sie bei jenen Organen und Geweben aber nicht. Man kann also durch Feststellung der Höchstzahl der T-Chromozentren die Ploidie einer Pflanze bestimmen. Dieses Verfahren, das im einzelnen eingehend beschrieben wird, ist leichter und schneller als das der Chromosomenauszählung. Geeignet sind folgende leicht zu präparierende Organe und Gewebe: Die Zellen der Kalyptra und die Epidermiszellen des Hypokotyls (beide für die Ploidiebestimmung beim Saatgut), ferner die Epidermiszellen des Stiels des Kotyledo und des Stielchens des Fiederblattes, weiter die subepidermalen Zellen der Innenseite des Kelchblattes, und schließlich — nur beiTr. hybridum — die Epidermiszellen des Stielchens der Einzelblüte.BeiTr. pratense kann man mittels der T-Chromozentrenmethode leicht Aneutetraploide auffinden, deren T-Chromozentrenhöchstzahl 3 oder 5 beträgt; ihr Vorkommen deutet darauf hin, daß die betreffende tetraploide Sorte noch nich frei von Meiosestörungen ist.Mit 7 TextabbindungenHerrn Professor Dr.Hans Bauer zur Vollendung seines 60. Lebensjahres gewidmet.     

The development of Hexarthra spp. in a shallow alkaline lake

In Neusiedler See, a shallow alkaline lake with fluctuating water level and salinity, four species of Hexarthra occur: H. mira, H. fennica, H. jenkinae (occasional) and H. polyodonta. The analysis of longterm data reveals a general phenological pattern which does not change from year to year. They first occur in May, develop a maximum in June/July, sometimes a second one in August/September and disappear in October. But the species succession is different in the various years, occasionally only one species (H. mira or H. polyodonta) being present. There is a fairly consistent relation between the chemical conditions and the prevalent species; an increase in salinity favours the development of H. polyodonta. Low temperature and wind generated suspended particles have a negative influence on the development of the Hexarthra populations. Smaller populations of Hexarthra are in a relation to the occurrence of Leptodora indicating predation pressure of the latter species. In Neusiedler See the Hexarthra populations seem to be controlled to a great extent by abiotic factors, but predation by Leptodora and most probably by young fish seems to play an important role too.     

Chromozentrenuntersuchungen

Zusammenfassung Bei 4 Arten der Angiospermenfamilie der Kruziferen wurde die Zahl der Chromozentren (Prochromosomen) einer größeren Anzahl junger 1n–, 2n–, 3n–, 3n+ 2–, 4n–, 8n–, 16n– und 32n-Ruhekerne aus verschiedenartigen primären Meristemen zahlreicher, in verschiedener Entwicklungsstufe stehender Pflanzen festgestellt (Tabelle 1 und 2). Die Kerne stammten aus unbehandelten Gameto- und Sporophyten von Diund Tetraploiden sowie aus colchicinierten Keimwurzelspitzen von Diploiden. Die Chromozentrenzahl aller untersuchten Ruhekerne stimmte mit deren Chromosomenzahl überein.     

Yeast cell surface display system for determination of humoral response to active immunization with a monoclonal antibody against EpCAM

Even though an immunogenic formulation of the murine monoclonal anti-EpCAM (epithelian cell adhesion molecule) antibody Mab 17-1A, has been shown to evoke a strong humoral immune response in both, monkey studies and early clinical trials, conventional anti-EpCAM ELISA could not identify anti-EpCAM immune response in relation to treatment with Mab17-1A. In contrast, usage of cellulose membranes prepared by SPOT technology presenting overlapping EpCAM peptides allowed the unequivocal determination of EpCAM related antibodies present in monkeys sera after immunization with IGN101. Based on such contradictory results, it was of high interest to compare obtained data to a different method for better assessment of their possible interpretation. Therefore, in the present studies, some EpCAM peptides, determined as reactive by binding of IgG isolated from sera of treated monkeys on membranes prepared by SPOT technology, were represented on yeast surface using the pYD1 yeast display vector system. Binding of biotinylated IgG from sera was detected with streptavidin–FITC and quantity of binding was determined by FACS measurement. Though using this completely different method, experiments with pre-immune and immune sera of four monkeys exemplarily are comparable to the results obtained by analysis with synthetic peptide arrays.     

Folding and refolding of proteins in chromatographic beds

The correct folding of solubilized recombinant proteins is of key importance for their production in industry. On-column refolding of proteins is mainly achieved by three methods: size-exclusion chromatography, ion exchange chromatography and affinity chromatography using immobilized metal chelates. The principles of these methods were first laid down in the 1990s, but many recent improvements have been made to these processes including sophisticated changes to the mobile phase composition and the recycling of aggregates to improve yield. Advances have also been made in the use of immobilized metal affinity chromatography and by mimicking the natural folding process with artificial chaperones.