We used monoclonal antibodies and immunohistologic examination of lymph nodes, to elucidate the pathogenesis of lymphosarcoma induced by, infection with bovine leukemia virus (BLV). The superficial cervical lymph nodes from 3 BLV-infected but apparently healthy sheep and 5 sheep with full-blown lymphosarcoma were examined. We also investigated the integration of bovine leukemia provirus by use of Southern blotting. In lymph nodes from sheep lacking clinical signs of infection, in which the provirus had been integrated at multiple sites in the genome, many large hypertrophic follicles were observed in the cortex. These follicles had germinal centers consisting of CD4+T cells and B cells that expressed surface IgM (sIgM) and major histocompatibility complex (MHC) class-II antigens, but not B cell-specific B2 molecule. The percentage of CD4+T cells in the cortex was significantly (P < 0.05) higher than that of the controls and sheep with lymphosarcoma. In all sheep with lymphosarcoma, the lymph nodes were completely destroyed by proliferating neoplastic cells, and in addition, small atrophic follicles, which consisted of normal B-cell marker-positive cells, were seen near the trabecula and the subcapsule. In these instances, neoplastic cells appeared to be a monoclonal population derived from a single CD5- B-cell lineage and to be classified as 2 types, CD5-CD4-CD8-B2+MHC class-II+sIgM+ and CD5-CD4-CD8-B2+MHC class-II+sIgM-. Moreover, CD8+T cells infiltrated diffusely throughout the tumorous lymph nodes apart from the atrophic follicles, and CD4+T cells were observed around atrophic follicles. Both types of T cells were small-size, normal lymphocytes with round and noncleaved nuclei, and were apparently non-neoplastic cells. In fact, after separation by use of a panning method, it seems that, in blood mononuclear cells from BLV-infected sheep without clinical signs of infection, but in lymphosarcomatous stages, the proviral genome was integrated only in B cells.

Bovine leukemia virus (BLV) was transmitted by horse flies, Tabanus fuscicostatus, from a cow with a lymphocyte count of 31,500/mm3 to goats and dairy calves. As few as 10 and 20 flies transmitted BLV to goats and calves respectively, but the minimal number of flies required to transmit the infection was not established. Groups of 150 and 100 T fuscicostatus transmitted BLV to beef calves from a cow with a lymphocyte count of 14,600/mm3. These results support a role for horse flies in the horizontal transmission of BLV.

Bone marrow fibroblast colony-forming units (CFU-F) were evaluated in cats experimentally infected with different isolates of FeLV. Cats infected with the Kawakami-Theilen isolate of FeLV (FeLV-KT) had progressive decrease in the number of CFU-F at 2, 4, and 6 weeks after infection. The number of CFU-F in FeLV-KT-infected cats ranged from 38 to 70% of the preinoculation CFU-F value. Of 3 cats with FeLV-KT-induced suppression of CFU-F, 2 developed fatal nonregenerative anemia. Cats infected with the Rickard isolate of FeLV (FeLV-R) had more moderate decrease in the number of CFU-F at 2, 4, and 6 weeks after infection. The number of CFU-F in FeLV-R-infected cats ranged from 62 to 82% of the preinoculation CFU-F value. The FeLV-R-infected cats did not become anemic.

The amphotropic murine leukemia virus (MLV) has been shown to infect mammalian species other than mice. If this virus infects and expresses genes in cells of livestock species (cattle, sheep, and pigs) it has potential for use as a vector to produce transgenic livestock. Because the gene-injection technique for producing transgenic animals is inherently inefficient, our laboratory was interested in identifying or constructing retroviral vectors capable of infecting livestock embryos. The infectivity of an amphotropic MLV-based vector for ovine, bovine, and porcine cells was tested. Experiments were also conducted to test the ability of the amphotropic MLV promoter, compared with known strong promoters, to express genes in cells from these species. Results indicated that amphotropic MLV infects and expresses genes efficiently in porcine cells and is, therefore, a potential vector for producing transgenic pigs. Infection was not detected in cells from adult bovine and ovine species; however, low levels of infection, with subsequent gene expression, were detected in cells derived from bovine embryos.

The specificity of serum antibodies for the polypeptides of bovine respiratory syncytial virus (BRSV) was examined, using sera obtained from feedlot and range cattle. Test results in sera from feedlot cattle indicated a 60% rate of seroconversion and 95% seropositivity to BRSV, associated with lack of clinical signs of indicative respiratory tract disease. Exposure to other common respiratory tract viruses also was high (greater than 92% to bovine herpesvirus type 1, bovine viral diarrhea virus, and para-influenza virus type 3). Test results in sera from range cattle indicated BRSV serpositive rates of 28% in calves, 49% in yearling cattle, and 70% in mature cows; clinical signs of respiratory tract disease were not observed in these cattle. Antibodies to BRSV in sera from cattle in both environments reacted predominantly with polypeptides of molecular weight 80,000 through 85,000, 40,000 and 28,000. Reactivity to a glycoprotein of molecular weight between 43,000 and 44,000 and to several glycopolypeptides of smaller molecular weight increased in serum specimens obtained from feedlot cattle between time of entry into the feedlot and slaughter.

A survey on the prevalence and distribution of antibodies to BLV was performed by the agar-gel immunodiffusion test over a period from 1983 to 1985. More than 2,407 serum samples were collected from Holstein cattle raised in the eastern part of Saitama prefecture where suburban dairy farm is operated. The average positive rate of this period was 4.9 %. The rates of reactive samples varied from 2.6 to 9.8 % among the age groups of cattle from younger than one year to 14 years of age. The positive rate increased gradually with age. The positive rates also varied widely from 0 to 21 % among areas surveyed. Furthermore, there were large differences in this rate among farms even in the same area. The results were interpreted and discussed in connection with the enzootic feature of BLV infection.

To determine the phenotype of target cells for bovine leukemia virus (BLV) infection in sheep, we analyzed blood lymphocytes from BLV-infected clinically healthy and leukemic sheep by used of monoclonal antibodies. In clinically healthy and leukemic sheep that were BLV-infected, the blood concentration of T lymphocytes was within normal values, but the number of B lymphocytes was increased in several cases. In addition, the number of blood lymphocytes expressing the BLV antigen correlated well with that of B lymphocytes. Double immunofluorescence staining demonstrated that lymphocytes expressing BLV antigens bore B-cell but not T-cell surface markers. Moreover, neoplastic cells in the lymph nodes of leukemic sheep were stained immunohistochemically with an anti-B monoclonal antibody but not with any of anti-T monoclonal antibody tested, indicating that tumor cells are of B-lymphocyte origin. Collectively, these results show that BLV antigen-positive cells obtained from BLV-infected sheep that have no clinical signs and BLV-induced lymphosarcoma cells belong to the B-lymphocyte lineage.