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A trial vaccine containing pseudorabies virus (PRV) glycoprotein gC as the main component showed excellent protection against virulent virus infection in pigs. Glycoprotein gC-rich antigen was prepared by heparin affinity chromatography from PRV-infected cell lysates. The preparations were mixed with mineral oil adjuvant as a water-in-oil emulsion. Six-week-old pigs were immunized twice at two-week intervals with trial vaccines containing 128,000, 12,800 and 1,280 HA units per dose of gC antigen. They were then challenged with a virulent PRV at day 7 after the final immunization. Neutralizing (NT) antibodies were produced with increase of antibody titers after challenge. Pigs immunized with 128,000 HA units per dose of gC survived and showed no virus shedding during the 2-week experimental period after the challenge. The role of cell-mediated immunity was examined using BALB/c mice, and induction of gC-specific cytotoxic T lymphocytes (CTLs) was detected by 51Cr release assay. From these results with mice, it is inferred that cell-mediated immunity, especially CTL, may play an important role in the effectiveness of our trial vaccine in addition to humoral immunity.

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.

A pseudorabies virus (PRV) glycoprotein-mixed vaccine was prepared by heparin-affinity chromatography from PRV-infected PK-15 cell lysates. In our previous study, the trial vaccine was induced protection with suppression of virus shedding in one-month-old pigs and generation of cytotoxic T lymphocyte (CTL) response in mice. In this study, the effect of the trial vaccine on suppression of both virus shedding and reproductive failure in pregnant sows was examined. Three sows were vaccinated twice until one week before mating. Each of them was infected intranasally with 10(6) TCID50 of PRV on day 28, 54, or 85 after mating, respectively. Three other sows were also mated and challenged at the same time as the respective control. The vaccinated sows produced virus-neutralizing antibodies. Sows with high level of VN antibody lowered the level and period of virus shedding after challenge. The maximum level of shed-virus titers in vaccinated sows were 10(1.25) to 10(3) times lower than controls. Vaccinated sows shed virus for 1 or 5 days, while controls shed for 8, 9, or 12 days. No abortion or stillbirth was observed in vaccinated sows during pregnancy. On the other hand, control sow challenged at a late stage of pregnancy showed abortion and stillbirth. The results obtained here indicate that our trial vaccine is effective to prevent reproductive failure by pseudorabies virus.

Cysts of the K strain of Toxoplasma gondii were administered orally to nine pregnant cats having no antibody to the organism at each of the three stages of gestation, namely early (14 days), middle (30 days), and late (40 and 51 days). Premature and weak or deformed fetuses were observed in the offsprings delivered by mother cats infected with Toxoplasma in the early or middle stage of gestation, while the cats infected at the late stage were delivered of apparently healthy fetuses. Mother cats developed parasitemia excreting Toxoplasma oocysts in their feces and formed the antibody to the organism after the oral administration. Toxoplasma was recovered from the organs or tissues of all fetuses delivered by the mother cats infected in the early stage of gestation. All kittens born healthy from the cats infected in the middle stage of gestation were free from Toxoplasma, but 80% of the premature kittens were found to be infected. Toxoplasma was isolated also from the organs of about half the offsprings born healthy from the mother cats infected in the late stage of gestation. These results indicate that congenital infection with Toxoplasma may occur commonly and cause damage to the fetuses when pregnant cats are infected orally with Toxoplasma cysts in the early or middle stage of gestation.