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1.
Peripheral blood lymphocytes (PBL) from normal and bovine leukemia virus (BLV)-infected cattle were prepared by density gradient technique and incubated with and without phytohaemagglutinin (PHA) and pokeweed mitogen (PWM). RNA synthesis was determined at different periods of incubation by 3H-uridine incorporation. PBL from BLV-infected cows with persistent lymphocytosis (PL) showed the highest spontaneous RNA synthesis. PBL from BLV-infected cows with normal lymphocyte counts synthesized more RNA than cells from normal animals. Decreased mitogen responses were observed in PBL from infected cows with PL in comparison to normal and BLV-infected cattle without PL. PHA and PWM did not show significant differences in their degree of stimulation of RNA synthesis.  相似文献   

2.
Bovine leukemia virus (BLV) RNA was demonstrated in peripheral blood lymphocytes treated with concanavalin A or phytohemagglutinin from a BLV-infected ox, and in lymphosarcoma cells cultured without mitogen from an enzootic bovine leukosis cow by in situ hybridization using biotinylated DNA probes.  相似文献   

3.
Peripheral blood lymphocytes (PBL) from cattle and sheep exhibited natural cytotoxicity on a fetal lamb kidney (FLK) cell line that was persistently infected with bovine leukemia virus (BLV) by 20-h 51Cr release assay. This cytotoxic activity was tested using PBL from normal cattle and sheep or BLV-infected animals. Although cytotoxic activity was also found in PBL from normal animals and from BLV-infected, but clinically healthy animals, the activity in PBL from animals with persistent lymphocytosis or leukemic animals was markedly decreased. The cytotoxic activity of PBL from 3 sheep sequentially tested before and during the disease was found to decrease gradually with the progress of the disease, and finally no cytotoxic activity was found at the time of death due to leukemia. These results suggest that the natural cytotoxic activity in PBL may have a role in immunosurveillance for progress of the tumor.  相似文献   

4.
Although bovine leukemia virus (BLV) is mainly associated with infections of B-lymphocytes, we have previously reported the statistically significant increase in the T-lymphocytes obtained from BLV-infected asymptomatic aleukemic (AL) cattle. In this report the presence of BLV provirus in the DNA of immunoaffinity purified T-lymphocytes from AL animals was assessed using a highly specific radiolabelled (32P) BLV-DNA provirus probe and solid phase DNA hybridization. The BLV provirus was found in the DNA of the peripheral blood mononuclear cells of all AL animals tested and three of the four purified T-lymphocyte preparations from these animals. The purified T-lymphocyte preparations used in this study contained less than 4% detectable B-lymphocytes. One animal had no detectable B-lymphocytes in the purified T-lymphocyte preparation and the DNA from these cells also gave positive hybridization results. The lymphocyte blastogenesis assay was then used as an indicator of the functional ability of lymphocytes from these BLV-infected AL cattle to respond to mitogenic stimuli. The responsiveness of lymphocytes from these animals to the mitogens concanavalin A (Con A), phytohemagglutinin (PHA), and pokeweek mitogen (PWM) was comparable to that of lymphocytes from BLV-negative animals when changes in 3H-thymidine uptake (c.p.m.) were used as measurement of mitogenic-induced blastogenesis. This indicated that infection of the T-lymphocytes by BLV does not appear to alter the overall response of the lymphocyte populations to mitogenic stimuli. High levels of spontaneous blastogenesis in the absence of mitogenic stimulation were observed for lymphocyte preparations of AL animals. The reason for this proliferation of lymphocytes is unclear; however, sera from these AL animals were found to contain a blastogenesis-augmenting factor(s) when added to lymphocytes from BLV-negative control animals in the presence of Con A, PHA and PWM.  相似文献   

5.
The humoral and cellular immunological reactivity of sheep were studied throughout the first 32 weeks following experimental infection with bovine leukemia virus (BLV). Seroconversion of BLV-inoculated sheep occurred within 4 weeks, but infection was not transmitted to contact control sheep. Despite the persistence of the viral infection, no differences were demonstrated in leukograms, serum IgG concentrations, humoral response to immunization with an irrelevant antigen (rabbit red blood cells), phytomitogen (Concanavalin A and Pokeweek mitogen)-induced lymphocyte blastogenesis, or chemical (1-chloro, 2-4 dinitrobenzene) skin contact hypersensitivity, between BLV-infected and uninfected contact control sheep. These results demonstrate the absence of a nonspecific immunosuppressive effect of BLV and further negate the influence of a generalized immunological deficit on the development of clinical disease in BLV-infected animals.  相似文献   

6.
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 use 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.  相似文献   

7.
Ten 2- to 4-month-old rabbits were inoculated subcutaneously with bovine leukemia virus (BLV)-infected bovine or sheep cells. By 6 weeks after inoculation all ten rabbits had converted to BLV antibody-positive, and BLV or BLV antigen was detected in lymphocytes from most of the rabbits tested, although there were few antigen-producing cells. Three rabbits showed continuous respiratory symptoms after infection and one died with pneumonia. Humoral immune responses against mouse serum were significantly suppressed in BLV-infected rabbits compared with non-infected control rabbits. The lymphocyte blastogenesis response was also suppressed in BLV-infected rabbits. At the time of necropsy, six rabbits showed pulmonary lesions; however, none of the BLV-infected rabbits had tumors during an observation period of over 1 year.  相似文献   

8.
This study was designed to determine the relative infectivity of lymphocytes and secretions from BLV-infected cattle with and without persistent lymphocytosis (BLV+PL+ and BLV+PL-). Ninety-seven sheep of mixed sex and age were assembled into 21 experimental groups. The recipient sheep were inoculated intravenously with serial dilutions of whole blood, saliva or nasal secretions from BLV+PL+ and BLV+PL- donor cows. Between 200 to 20,000 cells from single and mixed BLV+PL+ or single and mixed BLV+PL- donor cattle were used for inoculation. A very small number of BLV-infected lymphocytes (200 cells) was sufficient to induce BLV infection in sheep inoculated with diluted whole blood from BLV+PL+ cattle. The inoculation of whole blood (containing up to 20,000 lymphocyte cells) from BLV+PL- cattle did not induce BLV infection in recipient sheep. Saliva and nasal secretions also failed to bring about BLV transmission.  相似文献   

9.
The in vivo and in vitro effects of Trichinella spiralis excretory-secretory (ES) antigens on porcine peripheral blood lymphocyte (PBL) responses induced with mitogens (phytohemagglutinin, PHA; concanavalin A, Con A; pokeweed mitogen, PWM) or unrelated antigen (Protein A) were studied to determine whether ES antigens depress lymphocyte responses in experimental swine trichinosis, and/or if this response was manifested after lymphocytes from infected pigs had been pretreated with ES antigens. Additionally, the range of inhibition of lymphocyte responses was tested in parasite-free pigs using different doses of ES antigens and compared with the responsiveness of control cultures from the same animals. The responses of lymphocytes from pigs inoculated with 4 x 10(3) muscle larvae (ML) were strongly depressed (P < 0.05) at post-inoculation days (PID) 7 (after stimulation with PHA), 14, 35 (Con A or PWM), and 49 (PWM). At PID 56 and 63 the lymphocytes from T. spiralis-infected pigs responded better (P < 0.05) to all three mitogens than those from non-infected controls. After 7 weeks post-inoculation, PBL which were pretreated with 10 or 250 micrograms ml-1 of ES antigens showed significantly weaker (P < 0.05, P < 0.001) responses to PWM or PHA, respectively, than those from non-infected animals. The responsiveness of lymphocytes from both groups of pigs to Protein A was not affected by the pretreatment with ES antigens in vitro. The responses of lymphocytes from the parasite-free pigs induced by PHA, PWM or Protein A were strongly depressed (P < 0.01) after in vitro pretreatment regardless of the dose of ES antigens (5, 10, 15, or 20 micrograms ml-1) applied.  相似文献   

10.
The purpose of this study was to determine the effect of T-lymphocytes and phytohemagglutinin (PHA), a T-cell mitogen, on the expression of bovine leukemia virus (BLV) in cultured B-lymphocytes from BLV-infected cattle and sheep. Bovine B-lymphocytes were isolated by negative selection via complement-mediated lysis of T-lymphocytes. Additionally, bovine and ovine B-lymphocytes were positively selected using fluorescence activated cell sorting. Expression of BLV in cultured bovine and ovine B-lymphocytes occurred in the absence of T-lymphocytes and without PHA stimulation. The results of this study demonstrate that BLV replication in cultured B-lymphocytes is T-cell independent. This finding may have implications for the mechanism of viral latency within infected B-lymphocytes.  相似文献   

11.
The single intradermal comparative test was used with both avian and bovine tuberculin. Three cattle infected with bovine leukosis virus (BLV) were used as a source of infection. BLV-positive and susceptible animals were tuberculin tested alternately. Fifteen susceptible calves and 15 susceptible sheep were tested. A further three calves and three sheep were used as controls; the needles of the tuberculin syringes were deliberately contaminated with blood from the BLV-infected cattle, before being used in the test. Whereas all three calves and the three sheep inoculated intradermally with contaminated needles developed BLV infections, all of the other 30 animals have remained serologically negative to BLV for 10 months. Transmission of BLV with needles contaminated with BLV-infected blood was prevented by wiping the needles with absorbent cotton wool.  相似文献   

12.
The BLV-induced leukemia--lymphosarcoma complex in sheep   总被引：3，自引：0，他引：3  
Sheep are highly susceptible to BLV infection and can be infected via several different means (routes). In all inoculated animals, specific anti-BLV antibodies can be demonstrated 1 to 3 months post-inoculation (p.i.). Between 10 and 13 months p.i., a moderate but persistent lymphocytosis (PL) may be detected in about 50% of the infected animals. This hematological disorder may be, but is not necessarily, associated with the development of a lymphosarcoma and can (might) be interpreted as a true lymphoid leukemia. According to findings revealed by immunolabelling and mitogen stimulation of peripheral blood lymphocytes, BLV-induced PL appears to be a B-cell disorder. Induced lymphosarcoma appears in about 40% of infected sheep during the 6 years p.i. It too is of B-lymphocyte lineage. In vitro studies demonstrate that BLV antigen is expressed exclusively in B-lymphocytes. Yet, BLV expression is greatly stimulated in whole lymphocyte culture by the addition of T-cell mitogen. This same phenomenon occurs when the supernatant of stimulated T-lymphocyte cultures is added to isolated BLV-infected B-lymphocytes. This observation supports the hypothesis that, as is the case with other retroviruses such as HIV, BLV is able to use the regular activation machinery of the immune system for its own replication and transmission. It seems, therefore, that the leukemia-lymphoma complex in sheep may serve as an accurate experimental model for the study of the biological properties of retroviruses.  相似文献   

13.
Alterations in peripheral blood lymphocyte subpopulations were examined in bovine leukosis virus (BLV)-infected cattle using antibodies specific for differentiation antigens in conjunction with analytical flow cytometry. Animals considered to be aleukemic and lymphocytotic were included in the study. Significantly fewer numbers of circulating B-lymphocytes (surface Ig-positive) and T-helper lymphocytes (BoCD4-positive) were identified in BLV-infected aleukemic cattle compared to non-infected controls while no significant differences were established for T-cytotoxic/suppressor lymphocytes (BoCD8-positive). In contrast, BLV-infected animals with persistent lymphocytosis had elevated numbers of circulating B-lymphocytes with no significant perturbation in circulating T-lymphocyte subsets identified when compared as a group with the negative control cattle. Application of regression analysis to data from individual lymphocytotic cattle demonstrated a significant correlation between absolute numbers of B- and T-lymphocytes. Increased numbers of B-lymphocytes were correlated with increased numbers of T-helper and T-cytotoxic/suppressor lymphocytes.  相似文献   

14.
Bronchoalveolar washings were harvested from the excised lungs of 68 normal sheep of 3 different age groups (A: birth to 8 weeks; B: 6 months to 2 years; C: older than 2 years). Cells and fluid obtained were examined quantitatively and functionally. Fewer cells were present in the lavage fluids of Group A sheep compared with those of Groups B and C. Macrophages were the predominant cell type (70-80%) in all sheep, with lymphocytes being second in number. The lower proportion of lymphocytes in young sheep (Group A) was attributable to lack of B-lymphocytes. As a proportion of the lymphocyte population T-cells were in the majority (60-80%) in all sheep. The proportion of null cells was higher in young sheep than in adults. Pulmonary lymphocytes from sheep of all ages responded poorly to stimulation by the non-specific mitogens phytohaemagglutinin, concanavalin A, pokeweed mitogen and lipopolysaccharide of Escherichia coli. The proportion of neutrophils was higher in sheep over 2 years old compared with younger animals. Eosinophils were present in all age groups but their proportion was greatly increased in animals over 2 years of age. Basophils were absent in young sheep and were present in only low numbers in the lungs of adult animals. In young sheep (Group A) IgG was the predominant immunoglobulin. With age, the percentage of IgG in lung fluid decreased while that of IgA increased so that IgA became the predominant immunoglobulin in older animals (Group C). In sheep of all ages IgM was present in negligible amounts. The highest value of complement (C3) occurred in adult sheep (Group B). The total white blood cell counts and differential blood counts of all the sheep were within accepted normal ranges. As in the lungs, there was an age-associated reduction in the proportions of blood T-lymphocytes and an increase in B-lymphocytes. In contrast to lung lymphocytes, those from the blood of sheep of all ages exhibited a wide range of responses to mitogens. The lowest stimulation indices were observed in the oldest sheep (Group C). The results provide background data against which assessment can be made of changes consequent upon infection with respiratory pathogens.  相似文献   

15.
Expression of L-selectin was determined by single- and two-colour immunofluorescence on granulocytes, peripheral blood mononuclear cells (PBMC) and blasts of bovine origin by means of a monoclonal antibody IVA94 which recognizes bovine L-selectin (CD62L). Cells were separated from peripheral blood of healthy cattle and colleagues infected with bovine leukaemia virus (BLV). BLV-infected animals comprised lymphocytotic and non-lymphocytotic cows. L-selectin was expressed on 90-98% of granulocytes in all tested animals. The percentage of PBMC expressing L-selectin was lower in cattle with persistent lymphocytosis than in non-lymphocytotic or BLV-free cattle, and inversely correlated with lymphocyte counts. The ratio of B lymphocytes stained for L-selectin was significantly decreased from 60.2 +/- 1.9% in BLV-free cattle to 43.8 +/- 3.6 and 22.5 +/- 5.7% in non-lymphocytotic and lymphocytotic cattle, respectively. B-lymphocytes stained for L-selectin exhibited about 50% reduction in L-selectin expression in BLV-infected cattle compared with BLV-free cattle, as judged by the mean fluorescence intensity (MFI). The percentage of L-selectin-positive PBMC not bearing surface immunoglobulin M (predominantly T lymphocytes) was comparable in BLV-free and BLV-infected cattle. However, L-selectin expression on T lymphocytes was reduced (about 50%) in BLV-infected cattle, as judged by the MFI. We suppose that BLV infection results in a decreased L-selectin expression on lymphocytes, and accordingly, it may contribute to deregulation of the host immune system.  相似文献   

16.
The effects of aflatoxin B1 on responses of the peripheral blood lymphocytes (PBL) of 2 normal animals to phytohemagglutinin, concanavalin-A, and pokeweed mitogen and of the PBL of 2 Mycobacterium bovis-infected animals to phytohemagglutinin and purified protein derivative of M bovis (PPD) were studied. Aflatoxin concentrations of greater than or equal to 10 microgram/ml significantly suppressed the lymphocyte response of normal animals to the phytomitogens. Lymphocyte response of M bovis-infected animals to specific antigen PPD was significantly suppressed at aflatoxin concentration of 0.5 microgram/ml. Fifty- to 100-fold higher concentrations of aflatoxin were required to produce 50% suppression of lymphocyte response to phytomitogens, as compared with that produced to PPD.  相似文献   

17.
A simple ELISA is described for quantifying expression of bovine leukaemia virus (BLV) antigens in short-term cultures of peripheral blood lymphocytes (PBL) isolated from infected cattle. The PBL-ELISA demonstrated that antigen expression levels in infected cattle could vary by more than 50-fold. Inoculation of sheep with dilutions of lymphocytes from two BLV-infected cattle, differentiated in the PBL-ELISA by 50 to 100-fold, suggested that antigen expression levels were correlated with infectivity. Haematological data indicated that increased antigen expression in PBL cultures was associated with an increased number of circulating B-lymphocytes, irrespective of whether or not an animal had lymphocytosis. This supported the hypothesis that BLV-infected cattle that are PBL-ELISA positive are more infectious and may present a greater risk of transmitting the disease. The applicability of the PBL-ELISA to a field situation was assessed with 98 BLV-infected cattle from three commercial dairy herds with infection prevalences of 11%, 23% and 47%. Similar percentages (49%, 50% and 52%) of PBL-ELISA positive cattle were identified among those infected cattle available for testing in the three herds. An additional 22 infected cattle from an experimental herd were tested to assess the stability of antigen expression levels over an 8 month period. Fewer (27%) of these cattle were identified as PBL-ELISA positive and antigen expression levels were generally lower than those observed in the commercial herds. Antigen expression levels in the experimental herd remained stable over the period of the study. The potential of the PBL-ELISA to assist in BLV eradication programs by identifying those seropositive cattle with the greatest potential to transmit infection is discussed.  相似文献   

18.
Activation of dolphin peripheral blood lymphocytes by the phytomitogens, concanavalin A, phytohemagglutinin, and pokeweed mitogen, was evaluated. Whole blood culture technique was employed, which eliminated the necessity of lymphocyte isolation or purification procedures. Dolphin lymphocytes responded (by increased [14C]thymidine incorporation) to all 3 mitogens, but concanavalin A consistently produced the highest degree of stimulation.  相似文献   

19.
Immunological control of bovine leukemia virus (BLV)-infection has been reported as dependent on the expression balance of types 1 and 2 cytokines. In this report, mRNA expression of interferon (IFN)-gamma and interleukin (IL)-2 (type 1 cytokines), and of IL-4 and IL-10 (type 2 cytokines) were evaluated in concanavalin A-stimulated peripheral blood mononuclear cells (PBMC) from BLV-infected sheep. Despite the same dose of BLV-infection, the extent of viral propagation was markedly different between eight individual sheep by 12 weeks post infection. The virus did not propagate well in three sheep, which showed augmented mRNA expression of IFN-gamma, a strong indicator of cell-mediated immunity, immediately after BLV-infection. Among the other five sheep having more than 2% of BLV-infected cells among PBMC at 12 weeks post infection, four sheep developed B-cell leukemia or lymphoma within 2 years after infection. These observations indicate IFN-gamma expression may play an important role in the protective mechanism against BLV propagation at the early phase of the infection.  相似文献   

20.
Characterization of lymphocyte subpopulations in the bovine mammary gland was accomplished using cells obtained from dry secretions. Correlation of cell surface properties with functional capacity was attempted by assaying the ability to form erythrocyte-antibody (EA) rosettes, erythrocyte-antibody-complement (EAC) rosettes, and sheep erythrocyte (E) rosettes and the ability to respond to phytohemagglutinin (PHA), Concanavalin A (Con A), and pokeweed mitogen (PWM) in the lymphocyte stimulation test. Results were compared with those obtained for peripheral blood lymphocytes (PBL) from the same animals. Mammary gland lymphocytes (MGL) formed significantly fewer (p < .01) EA and EAC rosettes, but significantly greater (p < .01) E rosettes compared to PBL. MGL were significantly less responsive (p < .05) to mitogens than were PBL. MGL contained a large proportion of T lymphocytes, which do not respond to T lymphocyte mitogens in culture.  相似文献   

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