Hemagglutination with Equine Arteritis Virus.

Abstract

ABSTRACT. Equine arteritis virus (EAV) grown on RK 13 cell cultures was tested for hemagglutination (HA) with erythrocytes from a variety of species at 4°C, room temperature and 37°C. HA was observed at all temperatures with erythrocytes from mouse and chicken but not with those of cattle, horse, rabbit, guinea pig, mongolian gerbil, goose or chick embryo. Chickens showed an individual variation in agglutinability of their erythrocytes, requiring selection of birds to obtain erythrocytes for HA. The HA activity was enhanced by treatment of virus materials with Tween 80 followed by treatment with ether. The HA reaction was inhibited by specific antiserum. Higher HA-inhibiting (HI) antibody titers were obtained by the incubation of serum-HA antigen mixture at 4°C for 24 hr. HI antibody titers of individual horse sera showed a significant positive correlation with their neutralizing antibody titers. -KEY WORDS: equine arteritis virus, hemagglutination, hemagglutination-inhibition.J. Vet. Med. Sci. 59(10): 943-945, 1997 0.01% gelatin to make a 0.3% suspension. The ddY mouse erythrocytes were used, unless otherwise stated. The HA and HI tests were carried out by a microtiter method using 96-round-well microtiter trays (Sanko Junyaku Co., Tokyo, Japan) by the method described in our previous paper [5].Serum neutralization test (NT) was conducted by the 50% plaque reduction method using RK 13 cells and virus diluent containing 10% guinea pig serum (Denka Seiken, Tokyo) described in the previous paper [4].In preliminary experiments with the concentrated antigens prepared from RK 13 cell culture infected with the Bucyrus strain of EAV, erythrocytes from mouse were found to be agglutinated, although the HA titer obtained was low. To confirm this finding, 4 kinds of HA antigen were prepared and titrated for HA activity with mouse erythrocytes. As shown in Table 1, HA was observed with all HA antigens tested at room temperature. However, the results with the TE-cell HA and TE-fluid HA antigens were comparatively better than those with the non-TE-cell HA and non-TEfluid HA antigens. The highest HA titer of 64 was obtained with the TE-cell HA and TE-fluid HA antigens, and both the non-TE-cell HA and non-TE fluid HA antigens showed a considerably low HA titer of 16. All control HA antigens prepared from the mock-infected RK 13 cells treated with or without TE were HA negative (Table 1). In the following experiments, the TE-cell HA antigen was used, unless otherwise stated.TE-cell HA antigen was tested for HA with erythrocytes from a variety of species at 4°C, room temperature and 37°C. Mouse, ddY and BALB/c strains, and chicken erythrocytes were agglutinated at all three temperatures, whereas erythrocytes from horse, cattle, guinea pig, mongolian gerbil, goose and 10-day old chick embryo failed to be agglutinated under the test conditions (Table 2). In addition we found that erythrocyte samples from individual chickens showed a wide variation in their agglutinability, ranging from negative to positive, with the HA antigen.

Hemagglutination and Antigenic Comparison of Rabbit Hemorrhagic Disease Virus.

“…On the basis of the results, it is considered that the qualities of the erythrocyte's receptor against this virus might differ slightly for each blood group. On the other hand, it was reported that erythrocyte samples from individual animals, even when collected from the same species, showed a variation in their agglutinability [5,11]. These results are similar to ours, and may suggest that this phenomenon originates from the difference in the blood type of the individual animals from the same species.…”

Characterization of equine arteritis virus particles and demonstration of their hemolytic activity

“…It has been described that extracts prepared from EAV-infected cells, especially when they were treated with Tween=ether, can hemagglutinate chicken erythrocytes with titers up to 64 [10,13]. Our virus preparations (50 ml [1-10 mg=ml protein] incubated with 50 ml of 1% erythrocytes at room temperature) usually exhibited HA titers between 8 and 16.…”

Effect of Heparin on Hemagglutination by Equine Arteritis Virus.

“…Preparation of hemagglutinin: EAV hemagglutinin was prepared from the RK 13 cells infected with the Bucyrus strain of EAV by the method described previously [7] with the slight modification noted below. Briefly, the RK 13 cell monolayers in 500 ml bottles were infected with 10 2 TCID 50 of EAV.…”

“…The infected RK 13 cells were scraped off from the bottle surface with a rubber policeman. The cell-medium mixture was We have reported that equine arteritis virus (EAV), a member of the genus Arterivirus [9], agglutinates mouse and chicken erythrocytes [7]. In previous papers, the inhibitory effect of heparin on the growth or hemagglutination of the following viruses has been reported: herpes simplex virus (HSV) [10,12,15,16], bovid herpesvirus 1 (BHV-1) [14] and Aujeszkey's disease virus (ADV) [8,13] of the family Herpesviridae, subfamily Alphaherpesvirinae, genus Simplexvirus or Varicellovirus; bovine ephemeral fever, Kimberley and Berrimah viruses of the family Rhabdoviridae, genus Ephemerovirus [4,11]; Akabane and Aino viruses of the family Bunyaviridae [4,11]; and porcine reproductive and respiratory syndrome virus (PRRSV) [6].…”