Association between the strength of serologic recognition of bovine leukosis virus and lymphocyte count in bovine leukosis virus-infected cows

OBJECTIVE: To determine whether strength of serologic recognition of bovine leukosis virus (BLV) by use of ELISA is associated with blood lymphocyte counts. DESIGN: Prospective study. ANIMALS: 161 cows with positive results of ELISA for BLV. PROCEDURE: Sample-to-positive ratio (S:P), which is the ratio between the test sample and a positive control sample, was compared among lymphocytotic and nonlymphocytotic cows. A regression model was constructed to evaluate the association between blood lymphocyte concentration and S:P, age, and the interaction of these terms. RESULTS: Mean S:P differed significantly between lymphocytotic (2.58 +/- 0.36) and nonlymphocytotic (2.38 +/- 0.39) cows. Age and S:P were significantly associated with lymphocyte count. CONCLUSIONS AND CLINICAL RELEVANCE: Sample-to-positive ratio and lymphocyte count were related; however, cows with high S:P were not always lymphocytotic. Culling cows on the basis of S:P will reduce the herd load of infectious virus faster than random culling of ELISA-positive cows; however, culling on the basis of lymphocyte count will eliminate a greater proportion of the reservoir of infection.     

Mitogen and mixed lymphocyte culture responses of isolated bovine lymphocyte subpopulations

Examinations were made of the mitogen and mixed lymphocyte culture (MLC) responses of subpopulations of bovine lymphocytes isolated by density sedimentation following sequential E-rosetting with aminoethylisothiouronium bromide- and neuraminidase-treated sheep erythrocytes (EAET, EN). These procedures consistently separated 3 populations of E-rosetting T cells from a 4th population of non-E-rosetting cells (B and null cells). The isolated B and null cell population did not respond to phytohemagglutinin, concanavalin A, or pokeweed mitogen and responded minimally in mixed lymphocyte culture. Conversely, isolated T cells responded well to each of these stimuli. Moreover, differences in responsiveness were found among the 3 T-cell populations isolate by differential rosetting. T cells with receptors for both EAET and EN rosette formation were the most responsive to the mitogens used and demonstrated maximum activity in MLC. T cells with only EAET receptors had equivalent activity in MLC and less activity in response to mitogens. Cells with only EN receptors were the least active T-cell population in these assays. The different reactivities of these T-cell populations in lymphoproliferative assays indicated that they represent distinct subsets of bovine T cells.     

Factors affecting the infectivity of lymphocytes from cattle with bovine leukosis virus.

Peripheral blood mononuclear cells were obtained from 13 bovine leukosis virus infected cattle and inoculated subcutaneously into 29 recipient adult steers to determine (a) the number of mononuclear cells (equivalent amount of blood) necessary to cause infection and (b) factors influencing infectivity of mononuclear cells from bovine leukosis virus-infected animals. A total of 55 inoculations were made. Inoculation of 1 X 10(4), 2 X 10(4) and 5 X 10(4) mononuclear cells caused seroconversion in 12%, 57% and 62% of steers, respectively. No infections occurred with 1 X 10(3) or 2 X 10(3) mononuclear cells. Cattle infected for longer than 24 months and those animals greater than three years of age were more likely to cause infection with 1 to 5 X 10(4) mononuclear cells than were cattle infected for less than 24 months or animals less than three years of age. Lymphocytes from cattle with persistent lymphocytosis caused more infections when 1 X 10(4) or 2 X 10(4) mononuclear cells were inoculated, than did lymphocytes from nonpersistent lymphocytosis cattle; however, both groups were equally infectious when 5 X 10(4) mononuclear cells were inoculated. No differences were found in infectivity of experimentally vs naturally exposed animals.     

Response of cattle persistently infected with bovine virus diarrhoea virus to bovine leukosis virus

Six cattle persistently infected with bovine virus diarrhoea virus (BVDV) and seronegative, and two control, virus negative seropositive cattle were inoculated with lymphocytes infected with bovine leukosis virus (BLV). The two controls produced a normal immune response to BLV, developing antibodies at four and five weeks after inoculation. Two of the six cattle persistently infected with BVDV developed a strong antibody response by six weeks after inoculation with BLV. Four developed a depressed response to BLV, characterised in three by a 'hooking' reaction in the immunodiffusion test which persisted in successive bleedings but was interspersed occasionally by a weak positive reaction. In one of these animals, a series of 'hooking' reactions was followed by a number of negative results. The fourth animal remained serologically negative until 16 weeks after inoculation when a 'hooking' reaction was observed followed by a series of negative results. BLV was isolated from all the cattle persistently infected with BVDV at 42 or 58 weeks after inoculation regardless of whether the serum samples gave negative, 'hooking', weak positive or positive reactions in the immunodiffusion test. BLV was consistently isolated from the nasal secretions of a steer which was BVDV negative but seropositive. The possibility of decreased immune responsiveness to BLV in animals persistently infected with BVDV should be considered when formulating regulations governing the testing of animals for freedom from BLV.     

Inhibition of in vitro immunocyte function by sera from cattle with bovine leukosis

Most sera from leukaemic cattle inhibited phagocytic activity of normal bovine peripheral polymorphonuclear leukocytes, growth of interleukin 2-dependent bovine T cells and mitogen-induced (phytohaemagglutinin, concanavalin A, pokeweed mitogen, lipopolysaccharide and protein A) blastogenesis of normal bovine lymphocytes. By contrast, antibody-dependent, and spontaneous cell-mediated cytotoxicity were suppressed by only a few sera. The antibody titer against bovine leukaemia virus in these sera correlated with the percent inhibition of lymphocyte blastogenesis. These leukotic sera had no direct cellular cytotoxicity and the inhibitory activity was not lost by dialysis or heat inactivation at 62°C for 30 min. However, the activity was reduced by heating at 80°C for 30 min. Neither the concanavalin A sepharose 4B effluent fraction nor 3.5% polyethyleneglycol-treated serum was found to contain significant lymphocyte-inhibitory activity. Blastogenic transformation of lymphocytes prepared from leukaemic cattle was hardly detectable; however, the mitogen responsiveness of these lymphocytes was improved by a 37°C 1-h preincubation followed by washing.     

Changes in B cell and T cell subsets in bovine leukaemia virus-infected cattle

Direct immunofluorescence and fluorescence-activated cell sorter techniques were used for the detection of surface immunoglobulin positive (SIg+) cells in peripheral blood lymphocytes (PBL's) of bovine leukaemia virus (BLV) infected cattle with or without persistent lymphocytosis (PL+, PL-) and in BLV-free cattle. The percentage of SIg+ cells was more than twice as high in BLV+PL+ cattle than in BLV-free and BLV+PL- cattle. Bovine T cells, and T cell subsets were identified indirectly by the same techniques using three monoclonal antibodies (MAb's) specific for all T cells (IL-A43), T helper (BoT4) cells (IL-A12) and T cytotoxic (BoT8) cells (IL-A17). The major histocompatibility complex (MHC) determinants of both class II (BoT4) and class I (BoT8) as well as all T cells were significantly reduced in BLV+PL+ compared to BLV-free cattle. The actual decrease in the BoT8 cell subset or the dilution effect that would change effector:target cell ratio suggests that a resultant decrease in cytotoxic activity in BLV+PL+ cattle may play an important role in the progress of BLV infection in cattle.     

Frequency of latent bovine leukosis in Sweden.

The enzootic bovine leukosis occurs in Sweden especially in the south-eastern part of the country. An orientating serological survey was carried out on various categories of dairy cattle predominantly from this area. Serological and haematological examinations were also carried out on beef cattle herds connected to a volontary control program for bovine leukosis.     

Determination of proviral load in bovine leukemia virus-infected cattle with and without lymphocytosis

OBJECTIVE: To determine proviral load in bovine leukemia virus (BLV)-infected cattle with and without persistent lymphocytosis to assess the potential of transmitting the virus. ANIMALS: Cattle in 6 dairy herds. PROCEDURES: Blood samples from infected cows were evaluated 3 times at 6-month intervals for determination of proviral load via PCR assay, serologic results via ELISA, and hematologic status via differential cell counts. RESULTS: Infected cattle were classified into lymphocytotic and nonlymphocytotic groups. Lymphocytotic cattle consistently had > 100,000 copies of integrated provirus/mug of DNA (ie, high proviral load) in peripheral blood leukocytes. Titers of antibodies against BLVgp51 and BLVp24 indicated a strong immune response. Nonlymphocytotic cattle comprised 2 subgroups: a group with high proviral load and strong immune response, and a group with a weaker immune response, mostly against BLVp24, and a proviral load of < 100 copies/microg of DNA (ie, low proviral load). CONCLUSIONS AND CLINICAL RELEVANCE: Results emphasized the importance of characterizing nonlymphocytotic BLV-infected cattle during eradication programs. The risk of transmitting BLV infection from nonlymphocytotic cattle may differ depending on the proviral load. Nonlymphocytotic cattle with high proviral load could be efficient transmitters (as efficient as lymphocytotic cattle), whereas nonlymphocytotic cattle with low proviral load could be inefficient transmitters under standard husbandry conditions. Because most cattle with low proviral load do not develop anti-BLVp24 antibodies, it appears that lack of an anti-BLVp24 antibody response may be a good marker of this condition.     

Ultrastructural and immunologic characteristics of mouse x cattle xenogeneic hybridomas originating from bovine leukemia virus-infected cattle

Nine percent of xenogeneic hybridomas originating from a bovine leukemia virus (BLV)-infected cow secreted monoclonal IgM antibodies with multispecific reactivity. Similar reactivity was evident in some antibodies with an unusually long (> 50 amino acids) third complementarity-determining region of the heavy chain. Electron microscopy of hybridomas demonstrated the presence of c-type virus particles consistent with polymerase chain reaction detection of BLV env gene. Some hybridomas contained dilated rough endoplasmic reticulum and cisternae filled with moderately electron-dense granular substance compatible with plasma cells at presecretory stage. The number of chromosomes in xenogeneic hybridomas corresponded to the sum total of mouse and bovine chromosomes. None of the hybridomas showed polyploidy. The immunochemical and genetic analysis of stable bovine immunoglobulin-secreting xenogeneic hybridomas confirms that BLV infection causes polyclonal B cell activation regardless of antigen specificity. Presence of c-type particles in hybridomas suggests that T cell-derived cytokines are not required for sustained BLV expression.