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, Flaviviidae, 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.

Bovine blood lymphocytes, depleted of macrophages by absorption on plasma-gelatin coated plastic flasks, followed by passage through Sephadex G-10 columns, failed to respond to pokeweed mitogen stimulation. Adherent monocytes or alveolar macrophages added to purified lymphocyte preparations at 10% or less were able to restore the transformation response. Exposure of alveolar macrophages or purified lymphocytes to 2 bovine respiratory syncytial virus strains for 24 hours substantially reduced the transformation response when mixed with uninfected lymphocytes or macrophages. Exposure of alveolar macrophages or purified lymphocytes to 2 bovine parainfluenza type 3 virus strains produced a similar reduction in activity after 48 hours, Heat inactivation of the viruses removed their inhibitory ability. Immunofluorescence studies revealed that both alveolar macrophages and lymphocytes were permissive for parainfluenza type 3 virus, whereas only a small number of alveolar macrophages and lymphocytes were infected with respiratory syncytial virus. The results suggest that both viruses are capable of adversely affecting the interaction between macrophages and lymphocytes, although the mechanisms by which this is achieved may be different.

Pulmonary lavage samples were collected from 90- to 130-day-old calves before and 6 days after aerosol inoculation with bovine herpesvirus-1 (BHV-1) or parainfluenza-3 (PI3) virus. Alveolar lining material was separated from lavage fluids by high-speed centrifugation and phospholipids were extracted from alveolar lining material and analyzed by high-performance liquid chromatography. Phosphatidylcholine and phosphatidylethanolamine were 74.2 +/- 6.5% and 13.3 +/- 2.8%, respectively, of the total phospholipid content in the surfactant obtained from calves before virus inoculation. Other phospholipids were represented by substantially lower percentages. Infection with either of the 2 viruses caused a significant (P < 0.05) decrease in the percentage of phosphatidylcholine to 66.0 +/- 8.0% and 65.1 +/- 10.8% in the calves inoculated with BHV-1 and PI3 virus, respectively. A significant (P < 0.05) increase in the percentage of phosphatidylethanolamine to 18.1 +/- 2.2% and 17.8 +/- 4.5% developed in calves inoculated with BHV-1 and PI3 virus, respectively. Infection with BHV-1 also induced an increase (not significant) in the percentage of phosphatidylinositol from 5.5 +/- 2.8% to 7.8 +/- 2.2%. A similar, but not significant, increase in the percentage of phosphatidylinositol was also seen in the calves inoculated with PI3 virus. Less substantial changes in the percentage of other phospholipids were detected after virus infection.

In a search of viral agents in pulmonary macrophages of horses with chronic pulmonary disease, equine herpesvirus 2 was found to be unique. In 8 of 9 horses with chronic pulmonary disease, antigens of equine herpesvirus 2 were detected by indirect immunofluorescence staining of scattered foamy macrophages immediately after harvesting by bronchoalveolar lavage and fractionation on metrizamide gradients. In a healthy horse, antigens were not found. After 1 week of cultivation of bronchoalveolar lavage cells from a second group of 9 horses with chronic pulmonary disease, viral antigens were detected in 90% of the adherent pulmonary macrophages. In 2 of 3 healthy horses, viral antigens also were found in 90% of the adherent pulmonary macrophages. Antigens of equine herpesvirus 1, equine herpesvirus 4, parainfluenza virus 3, or adenovirus were not detected. Antigens of the 5 investigated viruses also were not detected in lung tissue slices from a third group of 14 horses, 4 healthy; 7 with varying degrees of bronchiolitis, 2 of which also had chronic intestitial pneumonia; 2 with eosinophilic bronchitis; and 1 with pulmonary hemorrhage. The exclusive presence of equine herpesvirus 2 in pulmonary macrophages was confirmed qualitatively by isolation of infective virus by cocultivation. In a fourth group of 12 horses with chronic pulmonary disease, infective virus could be isolated from pulmonary macrophages of 3 horses and from mixed-blood leukocytes of 5 horses. Virus isolations from 2 healthy horses were not successful from pulmonary macrophages, whereas 1 isolation was obtained from mixed-blood leukocytes. Other viruses were not detected by cocultivation.

The immune receptor-mediated functions of bovine alveolar macrophages (AM) inoculated in vitro with bovine herpesvirus-1 (BHV-1) or parainfluenza-3 (PI-3) virus were tested in the presence or absence of virus-specific antiserum or pulmonary lavage fluids collected from calves 6 days after inoculation with BHV-1 or PI-3 virus. The Fc and C3b phagocytic indices of noninoculated AM, collected from 6- to 16-week-old calves, ranged from 75 to 87 and 59 to 64, respectively, and the binding indices ranged from 5 to 8 and 22 to 28, respectively. Infection of AM with either BHV-1 or PI-3 virus had no significant effect on receptor-mediated phagocytosis or binding, with the exception of a significant (P < 0.05) decrease, from 64 to 46, of the C3b phagocytic index of PI-3 virus-infected AM. The addition of lavage fluids, collected after BHV-1 or PI-3 virus infection, to AM infected with the respective virus caused a significant (P < 0.05) decrease in phagocytic indices with values for the Fc and C3b indices in BHV-1-infected AM decreasing from 81 to 49 and from 47 to 8, respectively, and those for the PI-3 virus-infected AM from 79 to 51 and from 46 to 15, respectively. The binding indices of virus-infected AM increased with the addition of viral lavage fluids, but the only significant (P < 0.05) increase was for C3b binding in PI-3 virus-infected cells, which increased from 33 to 56. Virus-specific serum added to AM infected with the respective virus also caused significant (P < 0.05) decreases in the Fc and C3b phagocytic indices, with those for BHV-1-infected AM decreasing from 81 to 24 and from 47 to 5, respectively, and those for PI-3 virus-infected AM from 79 to 23 and from 46 to 3, respectively. The Fc binding index significantly (P < 0.05) increased with the addition of virus-specific serum from 8 to 34 and from 10 to 42 in BHV-1 and PI-3 virus-infected AM, respectively. The C3b binding index of these AM also increased, but not significantly. Infection of AM with either BHV-1 or PI-3 virus had no significant effect on the phagocytosis of opsonized (OPZ) or nonopsonized (nonOPZ) Staphylococcus epidermidis (SE). The addition of lavage fluids, obtained after BHV-1 infection, to AM infected with BHV-1, significantly (P < 0.05) decreased the percentage of phagocytosis of OPZ-SE from 28 to 21 and had a similar, but less substantial effect, on the phagocytosis of nonOPZ-SE. Lavage fluids collected after PI-3 virus inoculation, added to PI-3 virus-infected AM did not have a notable effect on the phagocytosis of OPZ-SE, but did cause a significant (P < 0.05) decrease in the percentage of phagocytosis of nonOPZ-SE from 25 to 17. The addition of virus-specific serum to infected AM caused significant (P < 0.05) decreases in the percentage of phagocytosis of OPZ-SE and nonOPZ-SE, with the values in the BHV-1-infected AM going from 28 to 11 and 16 to 9, respectively, and in the PI-3-infected AM from 36 to 12 and 25 to 13, respectively. Alveolar macrophages infected with either BHV-1 or PI-3 virus, in the presence or absence of lavage fluids from virus-infected calves or virus-specific serum, killed ingested SE as readily as noninfected AM. On the basis of the findings of this study, we suggest, as with other virus infections, that products of the host antiviral immune response interact with AM infected with BHV-1 or PI-3 virus or cause impaired internalization of receptor-bound particles, resulting in impaired AM antimicrobial functions.