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1.
Neonatally thymectomized and normal Holstein-Friesian calves were exposed to bovine viral diarrhea virus and challenged 22 days later. Baseline values in non-specific mitogen induced lymphocyte blastogenesis and serologic responses to tetanus toxoid and Brucella abortus strain 19 were established in neonatally thymectomized and normal calves.Neither viral recovery nor protective antibody production followed the very low level primary exposure to bovine viral diarrhea virus. The expected clinical response to an appropriate challenge inoculum did not occur, however, suggesting an immune response to bovine viral diarrhea virus had been initiated by the primary exposure. Five of eight calves developed protective serum levels of bovine viral diarrhea antibody. This may represent an example of immunologic priming. Exposure to low levels of bovine viral diarrhea virus also has potential implications in the pathogenesis of chronic bovine viral diarrhea wherein low level exposure may interfere with the development of protective levels of bovine viral diarrhea antibody as was observed in three calves.Circulating lymphocyte concentrations and results of non-specific mitogen stimulation of lymphocytes revealed a reduced phytohemagglutinin response in conjunction with a slightly increased response to bacterial lipopolysaccharide and pokeweed mitogen in the presence of moderate lymphopenia. The capacity of lymphocytes to respond to phytohemagglutinin was unchanged but an increased total response, as occurred in control thymectomized and intact calves, did not occur.The response to tetanus toxoid, a thymus-independent antigen, was increased following exposure to bovine viral diarrhea virus, perhaps reflecting an alteration in thymic-independent regulation. The lack of a differential response to Brucella abortus strain 19 implied competence in thymus-dependent helper cells.  相似文献   

2.
A microculture technique was developed for the in vitro blastogenesis of feline lymphocytes. Blastogenesis of ficoll-diatriazoate gradient separated mononuclear cell, washed blood and whole blood were compared. In general the whole blood cultures yielded higher stimulation indices (SI) than the washed blood or separated mononuclear cell cultures.The effect of several variables on the stimulation of lymphocyte cultures was examined. A cell concentration of 3 × 105 cells/well and a 1:20 dilution of washed and unwashed whole blood gave optimal stimulation with concanavalin A (Con A). Phytohaemagglutinin-P (PHA-P) did not give significant levels of stimulation. Inactivated fetal calf serum (FCS) at levels of 2.5% (for washed blood) and 5% (for separated mononuclear cell and whole blood) gave highest SI. Supplementation with FCS was preferable to autologous, homologous or horse sera for all cultures. Optimal SI was obtained in all cultures incubated for 3 days and labelled with 1 μCi tritiated thymidine (3H-TdR) for the last 16 hours. The highest SI were in the range of 70 to 105 (18,764 to 42,681 counts per minute (CPM) for separated mononuclear cell culture, 100 to 165 (28,403 to 45,334 CPM) for washed blood culture and 105 to 186 (41,076 to 69,999 CPM) for whole blood culture.  相似文献   

3.
Lymphocytes from healthy adult cats were separated into T and B cell-enriched subfractions by centrifuging rosetted cells on sodium metrizoate/Ficoll gradients. The responsiveness of unseparated lymphocytes (T + B), and T and B cell-enriched subfractions to stimulation with mitogens (phytohemagglutinin-P (PHA-P), concanavalin A (ConA), and pokeweed mitogen (PWM) was tested. Cultures of unseparated lymphocytes and those enriched in T cells showed similar responsiveness of PHA, ConA, and PWM stimulation; however, only a weak response to ConA and PWM was observed in B cell-enriched cultures. The mitogenic effects of PHA-P, ConA, and PWM on feline lymphocytes appeared to be due primarily to T-cell activation.  相似文献   

4.
Three hundred and three chicks of both sexes, from a synthetic dam line (SDL) of broiler chickens, were studied for economic traits (body weights at 4, 5 and 6 weeks of age) and immunological traits (humoral and cell mediated immune responses, and serum lysozyme concentration). The objective was to evaluate these traits and to estimate their genetic and non-genetic parameters. The humoral immune response was assessed by estimating the antibody response to sheep red blood cells using the haemagglutination (HA) test and serum IgG concentration using single radial immunodiffusion (SRID). The cell mediated immune (CMI) response was estimated as in vivo response to a mitogen (PHA-P). Serum lysozyme was measured by lysoplate assay. Least squares means for body weight at 4, 5 and 6 weeks were 684 +/- 20, 920 +/- 19 and 1205 +/- 28 g, HA titre was 6.289 +/- 0.246, CMI was 0.438 +/- 0.015 mm, lysozyme was 1.860 +/- 0.047 microg/ml and IgG was 6.287 +/- 0.194 mg/ml. There was an effect of sire on HA titre and on body weight at 4, 5 and 6 weeks of age; males were heavier than females. Heritability estimates were high for body weights but low for immunological traits. Phenotypic correlations (rp) among body weights were high and positive but were very low between body weights and most immunological traits. Among the immunological traits all rp were very low. Genetic correlations (rg) of body weights were positive and medium to high with CMI and HA and negative with serum IgG.  相似文献   

5.
We report here the development of a fatal lymphoproliferative disease in a common marmoset (Callithrix jacchus) following inoculation of the Ag876 strain of Epstein-Barr virus (EBV) and of the tumor promoting agent 12-O-tetradecanoylphorbol-13-acetate (TPA). The marmoset showed antibodies against EBV capsid and early antigens (VCA, EA) but not against nuclear antigen (EBNA). EBNA was detected in impression smears from lymph nodes and liver, and the presence of the EBV genome was detected from the same organs by hydridization techniques. To our knowledge this is the first report of the oncogenicity of this strain of EBV. The need for further studies to assess the in vivo interaction between EBV and tumor promoting agents in the development of malignancies is stressed.  相似文献   

6.
An experiment was conducted on 30 male Nellore lambs with average body weight (BW) of 15.45 +/- 0.06 kg to determine the level of the copper (Cu) supplementation in diet from inorganic and organic sources required for optimum immunity and its effect on copper dependent enzymes by allotting them randomly to five groups in completely randomized design. The dietary treatments were viz., basal diet (no Cu supplementation, BD), other four groups were offered BD supplemented with 7 or 14 ppm Cu from copper sulphate (CuSO(4)) and Cu-proteinate, respectively. The lambs were fed the respective diets at 3.5 per cent BW to meet the requirements except Cu for 180 days. The humoral immune response against Brucella abortus and chicken RBC was assessed after 90 days of feeding. The in vivo delayed type hyper sensitivity reaction against PHA-P and in vitro lymphocyte proliferation against Con A indicative of cell mediated immune response (CMI) was assayed at 180 days of feeding. At the end of experiment four lambs from each group were slaughtered for estimation of liver superoxide dismutase activity (SOD). The ceruloplasmin and RBC-SOD activities were higher (P < 0.05) in 14 ppm Cu supplemented lambs from Cu-proteinate at 90 and 180 days, while the liver SOD activity was higher (P < 0.05) in lambs fed 14 ppm Cu from CuSO(4). The STAT titres against B. abortus were higher in Cu supplemented lambs, with no effect of dose of supplementation. Lambs supplemented with Cu-proteinate had higher titers than CuSO(4) on 7 and 14 days of post sensitization. The total immunoglobulin concentration and the CMI response against PHA-P and Con-A was higher (P < 0.05) in lambs fed 14 ppm Cu-proteinae diet. The IgM level was though high in Cu supplemented lambs, no dose or source effect were observed. The study indicated that Cu dependent enzymes activity and immune response were highest and respond better against stress in lambs on 14 ppm supplemented Cu from Cu-proteinate.  相似文献   

7.
Optimum conditions for turkey lymphocyte transformation tests were determined. Thrice-washed turkey buffy-coat cells obtained after slow centrifugation (40 x g, 10 minutes) responded well to mitogenic stimulation. Turkey lymphocytes isolated on Ficoll-containing separation media largely lost their ability to respond to mitogens. Maximum responses were obtained with 2 x 10(7) lymphoid cells/ml. Responses to the mitogens were greatest when bovine fetal serum was used at a 2.5% concentration or pooled turkey serum and autologous plasma were used at a 1.25% concentration. Higher concentrations of turkey serum or plasma decreased the responses when sub-optimum doses of concanavalin-A (Con A) or phytohemagglutinin-P (PHA-P) were used. Serum-free cultures gave higher stimulation indices than cultures with serum only when sub-optimum doses of Con A or PHA-P were used. Optimum mitogen concentrations varied with individual birds, timing of the culture, temperature of incubation, and serum concentration in the cultures. Responses were usually greatest with final concentrations of 5 micrograms Con A/ml, 10 micrograms PHA-P/ml, and 20 micrograms pokeweed mitogen (PWM)/ml and when the cultures were incubated in 96-well microplates at 40 C in humidified air with 5% CO2 for 40-42 hours with pulsing with 3H-thymidine during the final 16 hours of incubation.  相似文献   

8.
The influence of moderate regular physical activity on the cell-mediated immunity was studied in growing pigs. Ten animals were subjected to physical training on a large animal treadmill, and 10 were kept in their pens throughout a 12-week experimental period. Regardless of whether the pigs underwent training or not, a whole blood lymphocyte stimulation test performed at 3 stages of the experiment revealed an equal ability of the cells to respond to stimulation induced by pokeweed mitogen and phytohaemaglutinin. The influence of serum from the pigs of the trained and untrained groups was studied in a stimulation test with purified mononuclear cells obtained from 2 healthy control pigs. The results indicated that no additional serum factors released by the physical training altered the blastogenic response of these lymphocytes. It is concluded that moderate exercise should not be regarded as a stressor which alters the cellular immunity in pigs.  相似文献   

9.
A Brucella abortus soluble antigen (BASA) preparation and fractions obtained thereof, by column chromatography, were compared in terms of their ability to induce specific lymphocyte stimulation responses (LSR) in lymphocytes from cattle infected with B. abortus. Endotoxin and protein contents and the fractions were determined. The LSR induced by BASA and the fractions were compared in terms of correctly identifying samples from infected and non-infected cattle. The sensitivity and specificity for each preparation were determined and these two attributes were then correlated with endotoxin and protein content. The results suggest that lymphocyte stimulation had greater association with relative protein content than endotoxin content of the antigen preparations.  相似文献   

10.
Phytohemagglutinin (PHA)-induced delayed-type hypersensitivity is an immunocompetent trait considered an indicator of cell-mediated immune or T-cell responses. Divergent selection was performed to generate high and low lines for response to PHA-P. Extreme-responder birds of the F2 generation in each line were used to study possible differences in macrophage activity and the associated functional genes. To evaluate macrophage activity, nitric oxide (NO) was estimated both systemically in serum and in in vitro monocyte culture. Semi-quantitative RT-PCR was used to detect the differential mRNA expression patterns of iNOS and MIP-1beta in monocyte culture, whereas T(H)1 cytokines (IL-2 and IFN-gamma) were studied in peripheral blood mononuclear cells (PBMC) at different time intervals after lipopolysaccharide (LPS) induction. The high line showed strong systemic, as well as in vitro NO production, compared to the low line, upon stimulation with NDV and LPS, similar to early and high iNOS mRNA expression. Following the pattern of iNOS gene expression, an early strong expression of cytokines with powerful iNOS-inducing action, such as IFN-gamma and the chemokine MIP-1beta, was observed in the high line. In contrast, for response to PHA-P, low expression of IL-2 was observed in the high compared to the low line. In conclusion, the study revealed that divergent selection for response to PHA-P resulted in a divergent effect on T(H)1 cell activity, resulting in altered macrophage function in chickens. Selection, based on response to PHA-P, could lead to more resistant birds or birds with an enhanced immune response.  相似文献   

11.
A colorimetric assay employing MTT (3-[4,5-dimethylthiazole-2-yl], 2-5-diphenyltetrazolium bromide) was used to determine the mitogenic response of intestinal intraepithelial lymphocytes (i-IELs) of chickens to T- and B-cell mitogens. Comparisons between mitogenic responses of i-IELs and peripheral blood lymphocytes (PBLs) were made to examine potential relationships. The results from this study indicated that T-cell mitogens, concanavalin A (Con A), and phytohemagglutinin-P (PHA-P) induced mitogenic stimulation in i-IELs. Although stimulation indexes of both i-IELs and PBLs were similar, the optical densities (ODs) of i-IEL cultures containing Con A or PHA-P were 20- to 50-fold lower than the ODs of PBL cultures containing the same mitogen. The lower conversion of MTT to formazan resulting in lower ODs in i-IEL cultures indicated a lower level of cellular activity in the i-IELs than in the PBLs. The mitogenic responses of both i-IELs and PBLs to Con A and PHA-P were dose dependent. The responsive concentration of Con A for i-IELs was within the range of 25-50 micrograms/ml, whereas the responsive concentration of PHA-P for i-IELs was 50 micrograms/ml. Three days of incubation was found to be adequate to induce a significant (P < 0.05) mitogenic response for both T-cell mitogens. Lipopolysaccharide was unable to induce a mitogenic response in i-IELs, which was attributed to the lack of B cells in the i-IEL population. This technique may prove useful in evaluating and studying the role of i-IELs in local cell-mediated immune responses of the gastrointestinal tract.  相似文献   

12.
A comparison of in vitro lymphocyte responses and delayed type tuberculin skin test responses was made in an animal experimentally exposed to a Mycobacterium bovis-infected animal and in cattle naturally infected with M. bovis. Tuberculin skin tests did not suppress in vitro lymphocyte responses to M. bovis PPD and to M. avium PPD tuberculin. The whole blood test used in these studies provided for considerable savings in time as compared to use of purified lymphocytes for evaluating in vitro cellular responses. Variations in the responsiveness of lymphocytes to specific mycobacterial antigens was observed, therefore, it is recommended that profiles be established using three or more tests conducted at 14-day intervals.  相似文献   

13.
The relationship between the optimum concentration of mitogen which induces lymphocyte blastogenic response and the receptor occupancy by mitogen was investigated. The receptor occupancies which induced maximal blastogenic activity in equine, bovine and canine peripheral blood lymphocytes (PBL) were 31.1 per cent, 26.5 per cent and 38.4 per cent with phytohaemagglutinin-P, and 48.2 per cent, 17.9 per cent and 24.5 per cent with concanavalin A, respectively. The data clearly show that each animal species had its own optimum concentration of mitogen for stimulation of PBL. Optimum concentration for blastogenesis and number of binding sites of each mitogen had a good correlation with each other for all three species.  相似文献   

14.
Cryopreservation of bovine peripheral lymphocytes and its effect on the in vitro response to concanavalin A tested in a microculture system is described. Using DMSO as cryoprotectant in the medium, the cells were cooled to ?30°C at 1.3°C/minute and further to ?80°C at 6°C/minute and then rapidly to ?196°C by dropping in liquid nitrogen. The cells were recovered by rapid thawing in water at 30–35°C and washed twice before use in the stimulation test. Ten percent DMSO had a much better protective effect than 5%; addition of 25% fetal bovine serum to the freezing had no favourable effect. In most of the 16 animals used in the experiments the frozen lymphocytes gave the same or a higher response to Con A than those kept in the DMSO containing medium at 4°C for two hours.The responses of the frozen cells were comparable to those of fresh lymphocytes (kept at 4°C for two hours in medium without DMSO).  相似文献   

15.
To test the hypothesis that variation in ability to respond immunologically correlates with health, Yorkshire pigs were bred for high (HIR) and low (LIR) antibody (Ab) and cell-mediated immune response (CMI). Selection was based on standardized measures of Ab (secondary response to hen egg white lysozyme, serum IgG concentration) and CMI (cutaneous delayed-type hypersenstivity to purified protein derivative of tuberculin after immunization with bacillus Calmette-Guérin and in vitro lymphocyte response to Con-A). Differences in Ab and CMI by line were not restricted to the antigens used in the selection. Antibody response to vaccines was highest in HIR and non-responders were restricted to LIR pigs. The HIR pigs had the best rate of weight gain. After infection with Mycoplasma hyorhinis, HIR developed more severe arthritis and less polyserositis. Differences were associated with variation in cytokine message in joint-related cells. Following exposure to attenuated transmissible gastroenteritis virus, natural killer cells of the LIR pigs but not of HIR or control lines, were unresponsive. Genetic selection for Ab and CMI may provide health and productivity advantages and complement traditional health-maintenance methods.  相似文献   

16.
Optimum conditions for the contact sensitivity (CS) test using 2,4-dinitrochlorobenzene (DNCB) were sensitization with 0.25 ml of a DNCB solution (10 mg/ml) and challenge with one-tenth of this dose two weeks later. This produced reaction in terms of skin thickness which could be easily observed and measured. The cutaneous hypersensitivity developed slowly, reaching its maximum manifestation after 24 h of challenge and gradually declining thereafter. The development of a skin reaction was typical of delayed type hypersensitivity and was characterized histopathologically by congestion, oedema, mononuclear and heterophilic cell infiltration in the dermal layer and lymphocytic perivascular cuffing. The efficiency of the CS test was compared with that of the lymphocyte transformation (LT) test and graft-versus-host (GvH) reaction for monitoring cell-mediated immunity (CMI) in normal and CMI deficient chickens. CMI deficient chickens were prepared by neonatal thymectomy and inoculation of antithymocyte serum. The CMI response of deficient chickens was significantly less (p less than 0.01) as determined by all three tests. The percentage decrease in response to mount GvH, CS and LT was 88.9, 58.3 and 74.6, respectively. All the tests were found to be equally effective in assessing CMI response in chickens as determined by comparing the means of their performances.  相似文献   

17.
Immunophysiological studies of interleukin-2 and canine lymphocytes.   总被引：3，自引：0，他引：3  
Interleukin-2-dependent pathways of lymphocyte activation were investigated in canine peripheral blood lymphocytes (PBL) following stimulation with T-cell mitogens including phytohemagglutinin, phorbol ester (TPA), calcium ionophore (ionomycin), and human recombinant interleukin-2 (hrIL-2). The ability of the stimulated cells to produce interleukin-2 (IL-2) was determined using murine indicator cell lines. IL-2 receptor expression by mitogen-stimulated canine PBL was confirmed by the binding of hrIL-2 with high affinity, and with characteristics comparable to those of the human and murine IL-2 receptor. Examination of serum and PBL from two dogs that were treated with hrIL-2 and human recombinant tumor necrosis factor for systemic mast cell tumors showed that in one dog, IL-2 could be measured in the serum. Concurrently, the in vitro mitogenic response of this dog's PBL to hrIL-2 occurred earlier, possibly reflecting an increase in the relative number of IL-2-responsive cells within the PBL population.  相似文献   

18.
Ó Nualláin, E.M., Davis, W.C., Fisher, A.D. and Monaghan, M.L., 1996. Development of a sandwich immunoassay for the detection of soluble bovine interleukin-2 receptor-α (sIL-2R-α) and its use to measure cell-mediated immunity in cattle. Veterinary Research Communications, 21 (1), 19-28. Stimulation of T-lymphocytes with mitogens or antigens results in the production of the cytokine interleukin-2, which exerts its physiological effect by interacting with a specific IL-2 receptor on the cell surface. The α-chain of this receptor is induced and expressed on the cell surface after lymphocyte activation. Following continuous antigen stimulation, a smaller soluble form of this α-subunit (sIL-2R-α) is shed from the membrane of activated cells. This study describes a sandwich ELISA for bovine sIL-2R-α that was developed using monoclonal antibodies specific for bovine IL-2R-α (CD 25). The feasibility of using sIL-2R-α released by activated T-lymphocytes as an in vitro marker of cell-mediated immunity (CMI) in cattle is demonstrated. Calves were immunized with the foreign protein keyhole limpet haemocyanin (KLH) and the development of CMI was followed using sIL-2R-α release, IFN-γ production and lymphocyte proliferation assay. The results showed that the release of sIL-2R-α by previously sensitized cells following stimulation with antigen is likely to be a useful marker of CMI in infectious diseases, and in the study of T cell antigens and/or novel vaccines. Using appropriate detection systems, the measurement of sIL-2R-α may also prove to be a useful marker of CMI in other species.  相似文献   

19.
The cell mediated immune response (CMI) was measured in calves after experimental infection with Mycobacterium avium. Using the tuberculin skin test a CMI response could be measured from four to 14 weeks after infection, and with a lymphocyte stimulation (LS) test from six to 40 weeks. One year after infection no CMI response was detected by either of the tests, in spite of the fact that in such calves M avium bacteria could be found in the intestinal lymph nodes at autopsy. After removal of mononuclear cells bearing receptors for the Fc part of IgG, the peripheral blood lymphocytes obtained from a calf infected one year earlier responded to M avium pure protein derivative in the LS test in contrast to lymphocytes obtained from uninfected calves.  相似文献   

20.
The tuberculin skin test is the conventional method of detecting infections with mycobacteria in animals. A positive reaction is considered to reflect cell-mediated immunity (CMI). CMI against mycobacteria can be studied by in vitro systems using suspensions of blood lymphocytes or leucocytes. The reactivity of these cells to different antigens can be measured in the lymphocyte stimulation (LS) (Muscoplat et al 1975, Bergman 1976, Johnson & Morein 1976), or leucocyte migration inhibition (LMI) (Aalund 1970, Clausen 1973) tests.  相似文献   

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