Die Präzipitationsreaktionen im Agargel bei der europäischen Schweinepest

Zentralblatt Für Veterinärmedizin Reihe B

2010

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EinleitungBei der Reaktion zwischen Pankreas oder Milz pestinfizierter Schweine und Schweinepest-Schweinehyperimmunseren konnten im Immunodiffusionstest und in der Immunoelektrophorese zwei serologisch nicht miteinander verwandte prazipitierende Schweinepestantigene nachgewiesen werden (6, 7). Beide Antigene wurden isoliert, physikalisch-chemisch sowie serologisch charakterisiert (3,4) und als Enzym(e) (1,4, 8, 10) identifiziert.Ergebnisse iiber den Nachweis von prazipitierenden Substanzen in konzentrierten Extrakten aus nichtinfizierten Organen (Pankreas, weii3e Blut-

Section: Introductionunclassified

Vergleichende Präzipitationsstudien zwischen hochvirulentem Virus der Europäischen Schweinepest, Schweinepestvirus-Spaltmaterial und den präzipitierenden Schweinepestantigenen aus den Organen pestinfizierter Schweine1

Zentralblatt Für Veterinärmedizin Reihe B

2010

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“…U. S. Copyright Clearance Center Code Statement: 0514-7166/81/2802-0l26~02.50/0 (1,15,16,19) or acquired by purchase (Behringwerke AG, Marburg/Lahn, FRG; Istituto Zooprofilattico, Sperimentale, Brescia, Italy; Pfizer S . A., Brussels, Belgium).…”

Section: Four Bovine Anti-bvdv Hyperimmune Sera (Bas-bvdv) and Six Pomentioning

confidence: 99%

“…For degradation and test procedures (immunoprecipitation in vitro, IDT, micro countercurrent immunoelectrophoresis -MCIE -, affinity chromatography on protein A-Sepharose), the viral preparations were mixed with Triton X-100, sonicated, and dialyzed as described before (14). Details of the methods and materials used for IDT, MCIE and the isolation of the immune-complexes have been described in previous papers (14,16,20). For affinity chromatography, 15 mg. protein A-Sepharose CL-4 B (Deutsche Pharmacia GmbH, Freiburg) was swollen, washed and resuspended in 1OOpl.…”

Section: Four Bovine Anti-bvdv Hyperimmune Sera (Bas-bvdv) and Six Pomentioning

confidence: 99%

Differences in Reaction Behaviour of Structural Polypeptides of Bovine Viral Diarrhoea Virus (BVDV) with Antisera against BVDV and Hog Cholera Virus (HCV)

Zentralblatt Für Veterinärmedizin Reihe B

2010

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Summary Immune‐complexes formed between BVDV structural polypeptides and bovine anti‐BVDV hyperimmune sera as well as porcine anti‐HCV hyperimmune sera by precipitation reactions and by affinity chromatography on protein A‐Sepharose were tested for antigens by SDS‐PAGE. The analyses showed differences in the reactivity of the two species of antibodies. In contrast to the BVDV antibodies, the HCV antibodies failed to react with p 34. Comparing the reactivity with the glycoproteins gp 57 and gp 44 of the viral envelope, the HCV antibodies revealed a more intense reaction with gp 44, whereas the BVDV antibodies reacted more intensely with gp 57. Zusammenfassung Unterschiedliche Reaktion von Strukturpeptiden des bovinen Virusdiarrhoe Virus (BVDV) mit Antiseren gegen BVDV und Schweinepestvirus (HCV) Immunkomplexe von BVDV‐Strukturpolypeptiden und Rinder‐anti‐BVDV‐Hyperimmunseren, sowie Schweine‐anti‐HCV‐Hyperimmunseren, wurden mittels Präzipitation und Affinitätschromatographie auf Protein A‐Sepharose hergestellt und im SDS‐PAGE‐Test untersucht. Die Ergebnisse zeigten Unterschiede in der Reaktivität der beiden verschiedenartigen Antikörper. Im Gegensatz zu den BVDV‐Antikörpern reagierten die HCV‐Antikörper nicht mit dem p 34. Bei dem Vergleich der Reaktivität mit dem viralen Hüllglykoprotein gp 57 und gp 44 zeigten die HCV‐Antikörper eine stärkere Reaktion mit dem gp 44, während die BVDV‐Antikörper stärker mit dem gp 57 reagierten. Résumé Réaction différente des peptides de structure du virus de la maladie des muqueuses (BVDV) avec des antisérums anti‐BVDV et anti‐virus peste porcine (HCV) A l'aide de la précipitation et de la Chromatographie d'affinité établi sur sepharose‐A proténe et dans le test SDS‐PAGE, on a examiné les complexes immunologiques des polypeptides de structure BVDV et des hyperimmunsérums anti‐BVDV de bovins et des hyperimmunsérums anti‐HCV de pores. Les résultats ont montré des différences dans la manière de réagir des deux anticorps hétérogènes. Par opposition aux anticorps BVDV, les anticorps HCV n'ont pas réagi avec p34. En comparant la manière de réagir avec la glycoprotéine d'enveloppe virale gp 57 et gp 44, les anticorps HCV ont réagi plus fortement avec gp 44 et les anticorps BVDV plus fortement avec gp 57. Resumen Reacciones diferentes de los péptidos estructurales del virus de la diarrea virósica bovina (BVDV) con sueros anti‐BVDV y sueros anti‐peste porcina (HCV) Se obtuvieron complejos inmunes de polipéptidos estructurales del BVDV y sueros hiperinmunes bovinos anti‐BVDV y de sueros hiperinmunes porcinos anti‐HCV mediante precipitación y cromatografía de afinidad sobre sefarosaproteína A, examinándose en la prueba SDS‐PAGE. Los resultados mostraban diferencias en la reactividad de ambos anticuerpos diferentes. En contraposición a los anticuerpos BVDV, no reaccionaban los anticuerpos HCV con la p 34. En la comparación de reactividad con las glucoproteínas cobertoras virales gp 57 y gp 44, mostraban los anticuerpos HCV una reacción más intensa con la gp 44, mientras que los anticuerpos BV...

“…determination in the SDS-PAGE. The rest was taken for affinity chromatography on protein A-Sepharose.For this purpose, bovine anti-BVDV serum (BAS-BVDV) and porcine anti-HCV serum (PAS-HCV) of naturally or experimentally infected animals (3,11,13) were dialyzed against PBS, adsorbed onto protein A-Sepharose CL-4 B (Deutsche Pharmacia GmbH, D-7800 Freiburg) after thorough washing and resuspension in PBS. Excessive and nonreacting proteins were removed and the beads then mixed with soluble antigen preparations and gently shaken for 30 min at room temperature.…”

mentioning

confidence: 99%

“…T E N buffer ( p H 7.5) with 2 O/o SDS and 1 O/o 2-mercaptoethanol the contents of the tubes were sonicated as mentioned, shaken at room temperature for 2-3 hours, heated in a boiling water-bath for 5 min and subjected to SDS-PAGE. (2,4,(11)(12)(13)16).…”

mentioning

confidence: 99%

Analysis of Soluble Bovine Viral Diarrhoea Virus Antigens and Serological Relationship to Virus Structural Glycoproteins

Zentralblatt für Veterinärmedizin Reihe B

1980

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IntroductionPapers demonstrating bovine viral diarrhoea virus (BVDV)-specific antibodies (10) in sera from recovered cattle by using "soluble" antigen preparations, or describing the production of vaccines against BVDV (7, 17) and studies of serological relationship (1-3, 8, 13, 18) between BVDV and hog cholera virus (HCV) obviously deal with soluble antigens which differ in their chemical and biological properties. Differences have been found with regard to composition (3, lo), stability (3, lo), and buoyant density (3, 7,9).The following studies investigate whether BVDV structural glycopolypeptides (1 5 ) display serological identity with the soluble antigen possessing nucleoside tri-and -diphosphase (XTPase) activity (2) and which is serologically related to HCV (SA) (1,3,13). Material and MethodsThe cytopathogenic BVDV strains Oregon C 24 V and NADL, propagated in primary calf testis cell cultures and labelled with 3%-methionine in some of these experiments, were purified as described (15). Crude soluble antigen preparations were obtained by pelleting cell debris and virus (SW 27, 26 000 rpm, 4 h, + 4 "C). High-molecular proteins were removed by 40 O/o (w/v) ammonium sulphate precipitation. Purified SA was isolated from crude preparations according to V A N AERT et al. (3).Abbreviations used: BVDV = bovine viral diarrhoea virus; H C V = hog holera virus; SA = soluble enzymatically active and HCV-related BVDV antigen; IDT = immunodiffusion test; IE = immunoelectrophoresis; MCIE = micro countercurrent immunoelectrophoresis; XTPasc = enzymatically active for nucleoside tri-and diphosphate digestion; mol. wt. = molecular weight; BAS-BVDV = bovine anti-BVDV serum; PAS-HCV = porcine anti-HCV serum. U. S.Subsequently, each preparation of the soluble antigens was mixed with Triton X-190(1 'Yo, w/v), sonicated in an ice-bath for 45 sec, incubated overnight at + 4 OC and sonicated once more. After exhaustive dialysis (3 days a t + 4 "C) against PBS with 0.1 O/o Triton X-100 (w/v) and low-speed centrifugation (4,000 xg, 30 min, + 4 "C) a portion of each sample was assayed for precipitating activities and mol. wt. determination in the SDS-PAGE. The rest was taken for affinity chromatography on protein A-Sepharose.For this purpose, bovine anti-BVDV serum (BAS-BVDV) and porcine anti-HCV serum (PAS-HCV) of naturally or experimentally infected animals (3,11,13) were dialyzed against PBS, adsorbed onto protein A-Sepharose CL-4 B (Deutsche Pharmacia GmbH, D-7800 Freiburg) after thorough washing and resuspension in PBS. Excessive and nonreacting proteins were removed and the beads then mixed with soluble antigen preparations and gently shaken for 30 min at room temperature. After an incubation period overnight at + 4 OC, the beads were extensively washed on a suction filter with PBS and finally once with double-distilled water, and then transferred into small plastic tubes. SDS and 2-mercaptoethanol-containing T E N buffer (pH 7.5) were added and the mixtures heated in a boiling water-bath and subjected to SDS-PAGE (15).Infecti...