ABSTRACT
Heparin inhibited hemagglutination (HA) by pseudorabies virus (PRV), but not HA by Akabane virus, bovine adenovirus type 7, Fukuoka virus, Getah virus, Japanese encephalitis virus, and parainfluenza virus type 3 belonging to the families Bunyaviridae, Adenoviridae, Rhabdoviridae, Togaviridae, Flavivi-idae, and Paramyxoviridae, respectively. The minimal inhibitory concentration of heparin required to inhibit 8 HA U of PRV ranged from 0.005 to 0.01 U/ml. Mouse erythrocytes failed to combine with the HA inhibitory factor of heparin. On the other hand, mouse erythrocytes treated with heparinase had greatly reduced agglutinability by PRV. Virus-heparin complex formation could be observed by sedimenting heparin with the virus particles.
Subject(s)
Hemagglutination Tests/veterinary , Heparin/pharmacology , Herpesvirus 1, Suid/drug effects , Animals , Erythrocytes/metabolism , Hemagglutination Inhibition Tests/veterinary , Heparin Lyase , Herpesvirus 1, Suid/metabolism , Mice , Polysaccharide-Lyases/pharmacology , Temperature , Viruses/drug effects , Viruses/metabolismABSTRACT
Pseudorabies virus hemagglutinin was readily adsorbed on mouse erythrocytes at 4, 22, or 37 degrees C, but not on cattle erythrocytes. The adsorbed hemagglutinin could not be eluted from the cells by resuspending in phosphate-buffered saline (PBS), by incubating at 37 or 50 degrees C, or by incubating in the presence of neuraminidase. The receptor on mouse erythrocytes for the hemagglutinin was inactivated by trypsin, but not by neuraminidase, sodium deoxycholate (DOC), potassium periodate (KIO4), dithiothreitol (DTT), 2-mercaptoethanol (2-ME) and formalin. The hemagglutinin was inactivated by trypsin, alpha-amylase, pepsin, DOC, KIO4, and ethylendiamine-tetraacetic acid (EDTA), but not by papain, beta-glucosidase, phospholipase C, neuraminidase, DTT, 2-ME, Tween-80, ethylether, chloroform, trichloro-trifluoroethane, beta-propiolactone and formalin, suggesting that the hemagglutinin active component involved glycoproteins. The hemagglutinin was stable at 37 degrees C for lower temperatures but not at 60 degrees C or higher. The hemagglutinin activity was resistant to ultraviolet irradiation, while the infectivity was very susceptible. The hemagglutinin and the infectivity were readily sedimented by ultracentrifugation at 48,000 x g for 3 hr. In rate zonal centrifugation of the preparation on a sucrose density gradient, the hemagglutination (HA) activity showed a sharp peak at 1.22 g/ml coinciding with the peak of infectivity. The HA activity in the peak fraction seemed to be structually associated with virus particles. After fractionation of the virus by Nonidet P-40, the HA activity was found only in the fraction of the envelope material, indicating that the hemagglutinin is situated in the viral envelop.
Subject(s)
Erythrocytes/metabolism , Hemagglutinins, Viral/chemistry , Herpesvirus 1, Suid , Adsorption , Animals , Cell Line , Centrifugation, Density Gradient , Erythrocytes/drug effects , Hemagglutinins, Viral/drug effects , Hemagglutinins, Viral/radiation effects , Temperature , Ultracentrifugation , Ultraviolet RaysABSTRACT
Slow-reacting, complement-requiring hemagglutination-inhibiting (HI) antibody was detected in sera from pigs infected with pseudorabies virus; approximately 16 hemolytic units of complement were necessary for the detection of such antibody. Higher HI antibody titers were obtained when antigen and serum were allowed to incubate before addition of complement than when all three components were incubated at the same time. A HI test was developed in which antigen-serum mixtures were incubated at 4 degrees C for 48 h and then with complement at 37 degrees C for 1 h; this gave an improved sensitivity over the previous incubation without complement.
Subject(s)
Antibodies, Viral/analysis , Complement System Proteins , Hemagglutination Inhibition Tests/methods , Herpesvirus 1, Suid/immunology , Pseudorabies/diagnosis , Animals , Antibodies, Viral/immunology , Cells, Cultured , Guinea Pigs , Hemagglutination Tests , Predictive Value of Tests , Pseudorabies/immunology , Serologic Tests , SwineABSTRACT
Pseudorabies virus grown in CPK cell cultures was tested for hemagglutination (HA) with erythrocytes of a variety of species at 4 degrees C, 25 degrees C and 37 degrees C. HA was observed at all temperatures with mouse erythrocytes but not with cattle, sheep, goat, swine, cat, rabbit, guinea pig, rat, mongolian gerbil, chicken, and goose erythrocytes. Mice showed a strain variation in agglutinability of their erythrocytes, requiring selection of mice to obtain erythrocytes for HA. The HA reaction was inhibited by specific antiserum. Some factors involved in the HA and HA-inhibition (HI) were investigated and standard HA and HI tests were established. HI antibody titers of individual pig sera showed a significant positive correlation with their neutralizing antibody titers.
