首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献，搜索用时 873 毫秒
1.
The proliferation responses of antigen-specific lymphocytes from various anatomical sites were studied in dairy goats locally immunized with heat-killed Staphylococcus aureus (HKS). Animals were inoculated three times subcutaneously in the right udder with HKS at 1 month intervals. One week following the last inoculation, prescapular, mesenteric and ipsilateral (draining) and contralateral (non-draining) suprammammary lymph nodes were collected and the cells assayed in 3- and 6-day cultures to determine the immune proliferative responses of antigen-specific lymphocytes to HKS and the polyclonal T cell mitogen phytohemagglutinin (PHA). The cells from draining and non-draining supramammary lymph nodes responded to HKS in 3-day cultures. Peripheral lymph nodes, such as the prescapular, showed similar responses. In contrast, mesenteric lymph nodes responded optimally in 6-day cultures, notably to lower concentrations of the antigen. Cells from all lymph nodes tested showed increased responses to PHA in immunized animals, although non-draining lymph nodes demonstrated a greater response to the T cell mitogen than those of draining lymph nodes. These results suggest that unilateral introduction of Staphylococcus cell antigens to the supramammary region can induce an anamnestic response in ipsilateral as well as contralateral supramammary lymph nodes and other distant peripheral lymphoid organs. Furthermore, these data indicate that cells from intestinal lymph nodes respond differently from those of peripheral lymph nodes, suggesting the presence of a unique gastrointestinal lymphoid cell circulation in goats. Concomitant peripheral responses may be attributed to memory cell migration or to antigen leakage and relocation to distant sites from the inoculated region. Analysis with PHA suggests a difference in general responsiveness and perhaps, immunocompetence, by lymphocyte populations in various lymphoid tissues of immunized animals.  相似文献   

2.
Six day in vitro cultures of canine mononuclear leukocytes with or without prior carbonyl iron depletion were established with 5 commonly employed mitogens. Cultures were assessed for B-cell differentiation by induction of cytoplasmic and supernatant released IgG, M and A and T-cell differentiation by expression of Thy-1 antigen on the cell surface of lymphoblasts. The mitogen concanavalin A was shown to be a restricted T-cell mitogen, whereas pokeweed mitogen and protein A from Staphylococcus aureus stimulated maximal B-cell differentiation. These data establish the mitogenic specificity for canine lymphocytes in unfractionated peripheral blood leukocyte cultures and will permit an in vitro evaluation of various substances upon T- and B-lymphocyte functions.  相似文献   

3.
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   

4.
A sensitive limiting dilution analysis (LDA) was used to quantitate the local and systemic cellular immune response of cattle after immunization with keyhole limpet hemocyanin (KLH) and infection with Ostertagia ostertagi. The assay measures the proliferative response of bovine T-cells after in vitro stimulation with antigen. Interleukin 2 activity was supplied by supernates from mitogen-stimulated bovine peripheral blood lymphocytes (PBL) and accessory cell function was in the form of irradiated autologous PBL. The assay measures the response of a single cell and was most easily demonstrated in the lymph nodes draining the site of antigen inoculation. Comparison of cell frequencies and maximal responses generated in conventional proliferative assays showed several differences between the two assays. First, after antigen injection, the highest cell frequencies were seen in the draining lymph nodes within 3 days, and decreased by 10 days post-immunization. In contrast, in mass cultures maximal stimulation was not seen until 7-10 days after injection, but remained high up to 4 weeks after immunization. Second, at 17 days post-infection, a time of eruption of the parasite from the gastric glands, high frequencies of inducible cells were demonstrated by LDA in all lymphoid populations tested. In contrast, low levels of proliferation were seen in mass cultures. The reasons for these differences may include different sensitivities to suppression or more stringent requirements for specificity between the two assays. Finally, it was found that immunologically naive calves have relatively high frequencies of Ostertagia-specific cells in PBL, and that after infection these frequencies decrease. These results indicate either active suppression of the potential anti-Ostertagia response or an extra-vascularization of these cells to the site of infection.  相似文献   

5.
Mitogenic and antigenic lymphocyte stimulation were examined in Aujeszky's disease virus (ADV) infected pigs and in pigs vaccinated with modified live ADV. Neither infection nor vaccination had any effect on lymphocyte responsiveness to phytohaemagglutinin (PHA), pokeweed mitogen (PWM) or concavalin A (Con A). ADV antigen-responsive lymphocytes began to appear in the peripheral blood between 7 and 14 days after inoculation and could still be demonstrated in blood and spleen of infected pigs at 174 days after infection. In vaccinated pigs, sensitized peripheral blood lymphocytes could be detected up to at least 35 days after revaccination. Pre-incubation of ADV antigen with specific antibody markedly reduced lymphocyte stimulation. Non-immunized pigs showed no lymphocyte response to ADV antigen. Infected pigs exhibited no lymphocyte reactivity against antigens of non-infected cells.  相似文献   

6.
Lymphocytes of lambs on a low selenium/vitamin E diet were isolated from peripheral blood, and mitogenic responses to phytohaemagglutinin (PHA) tested in the presence of different doses of sodium selenite and emulsified vitamin E added in vitro. An enhancing effect of selenium was observed at doses of 1 ng/ml or less, and reached a plateau at about 10 ng/ml. Toxic effects were evident beyond 1 micrograms/ml. The stimulatory potential of selenium among lambs was inversely related to their ability to respond to PHA in control cultures but was not related to the blood glutathione peroxidase activity of the animals concerned. Optimal doses of vitamin E added to culture (0.15-1.5 micrograms/ml) elevated responses beyond those seen with selenium, but synergistic effects were not apparent. Similar results were obtained when lymphocytes from deficient, myopathic lambs were cultured with serum from lambs supplemented in vivo, and when PHA responses of untreated and treated lambs were compared. Tests with other phytolectins (concanavalin A and pokeweed mitogen) suggested that the two micronutrients exert a differential influence on lymphocyte sub-populations. It was also concluded that the poor lymphocyte responses seen in myopathic lambs can be readily and rapidly reversed by injection of these nutrients, and that prophylaxis is most effective during the first 6 weeks of life.  相似文献   

7.
Several experimental parameters were examined to determine optimal conditions for proliferative responses of mammary mononuclear cells (MMC) obtained from six nonlactating dairy cows. These parameters were: pre-incubation of cells in medium prior to assay, mitogen concentration, assay incubation time, and type of culture medium. Response variables included viability of cells and the rate of proliferation as assessed by tritiated thymidine incorporation. Pre-incubation of cells in medium had no effect on the proliferative response of MMC. Whereas Concanavalin A (ConA; 3.3 or 6.6 micrograms/ml) and phytohemagglutinin (PHA; 1, 5, 10 micrograms/ml) did stimulate proliferation of MMC, the higher doses did not stimulate greater proliferation than the lower doses of mitogens. The greatest mitogenic response was obtained on days 2 and 3 of incubation. Proliferative responses were significantly higher at all mitogen levels tested in a 50-50 mixture of Rosewell Park Memorial Institute medium 1640 and Liebovitz-15 medium (RPMI/L-15) than in RPMI alone. Viability of MMC was also significantly higher in the RPMI/L-15 medium. To test whether the significant effect of media on blastogenesis was specific for mononuclear cells from the bovine mammary gland, peripheral blood lymphocytes (PBL) from four dairy cows were cultured with ConA and PHA in a mitogen assay in both RPMI and RPMI/L-15. Viability was measured on day of collection and on all culture days. PBL were stimulated equally in both media. PBL viability decreased significantly on day 1 in both RPMI and RPMI/L-15. These results suggest that the optimal culture conditions for blastogenic responses of mammary mononuclear cells and peripheral blood lymphocytes may differ.  相似文献   

8.
The effect of viable Mycoplasma bovis on the in vitro bovine peripheral blood lymphocyte response to phytohemagglutinin (PHA) was studied. Results showed that M. bovis did not act as a mitogen for bovine lymphocytes. Viable M. bovis produced a dose and time dependent suppression of the PHA stimulated lymphocyte response. Suppression was not a result of differences in the viability of infected or control lymphocyte cultures. The suppressive effect of M. bovis was found to be independent of the concentration of PHA used in the test and the lymphocyte response could not be restored by supplementation of the culture medium with arginine. Delay for 48 h after PHA stimulation before adding M. bovis to the lymphocyte cultures diminished, but did not prevent, the suppression of the lymphocyte response. These results show that suppression of the lymphocyte response does not require the presence of M. bovis during the period of PHA stimulation, and that M. bovis was capable of interrupting [3H]-thymidine incorporation in lymphocytes which were actively synthesizing DNA.  相似文献   

9.
In vivo inoculation of three-month-old calves with sodium diethyldithiocarbamate (DTC), killed Corynebacterium parvum or mycobacterium cell wall extract (MCWE) resulted in an enhancement of in vitro peripheral blood lymphocyte blastogenic responses to mitogens phytohemagglutinin (PHA) and Concanavalin A (Con A) in the first three days after treatment. In a separate experiment, blood lymphocytes isolated from a healthy nontreated calf were incubated in vitro in presence of each of the same immunostimulating agents and tested for their blastogenic responses to PHA and Con A. The results showed that all immunostimulants, excepting DTC, enhanced the in vitro blastogenic responses of lymphocytes to PHA and Con A. Finally, addition of MCWE to cultures of blood lymphocytes isolated from calves vaccinated intramuscularly with bovine rotavirus and adjuvant resulted in an enhancement of the in vitro lymphocyte transformation to rotavirus. Our study demonstrated that DTC, killed Corynebacterium parvum and mycobacterium cell wall extract were able to enhance bovine T cell proliferation in vitro.  相似文献   

10.
为获得金黄色葡萄球菌特异性抗原蛋白并以此制备多克隆抗体,应用DNAStar软件筛选金黄色葡萄球菌FnbpA、ClfA、Ebps的抗原表位,以柔性氨基酸序列连接后进行全基因合成,将目的基因克隆到pGEX-4T-1原核表达载体,转化至大肠杆菌感受态细胞,诱导表达,采用Sepharose 4B柱亲和层析纯化表达蛋白,制备抗原后对家兔进行皮下多点注射,于第42天采集外周血,获得血清。结果表明,本试验成功构建了金黄色葡萄球菌抗原基因表达载体,并获得了高免血清,经ELISA的方法检测血清效价可达到1∶10000以上,成功制备多克隆抗体,为以后采用免疫学方法快速检测金黄色葡萄球菌奠定了基础。  相似文献   

11.
Bovine fetuses were inoculated with Mycobacterium bovis, bluetongue virus or placebo at approximately 125 days of gestation, and blastogenic responses of peripheral blood lymphocytes and lymph node cells were determined at various time intervals after inoculation. Lymphocytes from all fetuses were stimulated by phytohemagglutinin, concanavalin A, and pokeweed mitogen, and peripheral blood lymphocytes gave consistently greater stimulation indices than did prescapular lymph node cells. Bluetongue virus infection did not consistently suppress mitogen induced lymphocyte blastogenesis. Lymphocytes taken from fetuses at 20 or 50 days after Mycobacterium bovis inoculation were not stimulated by purified protein derivative (PPD), whereas lymphocytes taken from adult cattle at similar intervals after Mycobacterium bovis inoculation were stimulated by PPD. Although lymphocytes from bovine fetuses may be stimulated by mitogens, antigen specific blastogenesis to a known inducer of cellular immunity was not detected by 175 days of gestation.  相似文献   

12.
Ten parasite-free lambs were drenched with oxfendazole on days 0 and 28 and, one day after each drench, were injected with human erythrocytes and ovalbumin. Ten other antigen-injected lambs were not drenched (controls). Lymphocytes collected 3 days after each antigen injection and cultured in RPMI 1640 plus 5% fetal calf serum (FCS) and lymphocytes collected 3 days after the first and 3 and 7 days after the second antigen injection and cultured in 50% autologous serum had decreased blastogenic activity compared with control lymphocytes. After the second drench, decreased blastogenesis was seen with lymphocytes collected on days 3 and 7 and cultured in 5% FCS and concanavalin A (Con A) and on day 3 when cultured in 5% FCS and phytohaemagglutinin (PHA). Decreased blastogenesis was also seen with lymphocytes collected 7 and 29 days after the second injection of antigen and cultured in 50% autologous serum plus Con A and on days 3, 7 and 29 when cultured in 50% autologous serum and PHA.Significantly depressed antibody responses to both antigens were seen after the second drench. The serum complement level was depressed 3 days after the second injection of antigen. Serum nitric oxide levels were significantly depressed 3 and 21 days after the first and 7 and 21 days after the second injection of antigen. There were no differences in levels of growth-promoting hormones but the drenched lambs gained significantly more weight than the controls.Abbreviations C complement - Con A concanavalin A - cpm counts per minute - EIA enzyme immunoassay - FCS fetal calf serum - IGF insulin-like growth factor - oIGF-1 ovine insulin-like growth factor-1 - PBS phosphate-buffered saline - PHA phytohaemagglutinin  相似文献   

13.
Eight lambs were experimentally infected with bovine respiratory syncytial virus (BRSV) and the responses of their peripheral blood lymphocytes to the mitogen phytohaemagglutinin and BRSV antigen compared with that of control lambs injected with tissue culture fluid. The lymphocyte transformation responses to phytohaemagglutinin were significantly reduced five and 10 days after experimental infection with BRSV (P less than 0.05). It appears that these reductions were associated with CD4+ lymphocytes because CD4-enriched lymphocytes obtained five days after infection had more significantly reduced responses to phytohaemagglutinin than those obtained from the same group before infection and from the control group five days after inoculation (P less than 0.01). There were no significant lymphocyte transformation responses to BRSV antigen in both groups of lambs up to 21 days after inoculation (P greater than 0.05).  相似文献   

14.
Chickens were inoculated wih the pathogenic Edgar strain of infectious bursal disease virus at 1 week, 2 weeks, or 1 day of age. In the 3 experiments, phytohemagglutinin stimulation of peripheral blood lymphocytes was significantly decreased on day 3 or 4 after inoculation. Subsequently, on days 7 through 21, stimulations were similar between lymphocytes from inoculated birds and those from control birds. Pokeweed mitogen stimulation was affected minimally in virus-inoculated chickens. In each experiment, on day 7, the spontaneous [3H]thymidine uptake was greater in nonstimulated lymphocyte cultures from inoculated chickens than in such cultures from control chickens. In an additional experiment, chickens 1 week of age were exposed to a pathogenic vaccinal virus given in their water. The vaccinal virus exposure resulted in significant decrease of phytohemagglutinin stimulation of lymphocytes on days 3 and 7 of the experiment. A significant decrease in pokeweed mitogen stimulation was observed on day 10 after inoculation.  相似文献   

15.
Twelve 9-week-old calves were divided into four groups; two groups were maintained helminth-free as controls and the other groups were given Ostertagia ostertagi infective-stage larvae (L3) orally. One group received 100,000 L3 as a single inoculum and the other group received L3 in increasing dosages at weekly intervals for 8 consecutive weeks. The blastogenic responses to concanavalin A (Con A), phytohemagglutinin (PHA), and a soluble larval antigen from O. ostertagi (SLA) of peripheral blood lymphocytes were evaluated using tritiated thymidine incorporation into DNA as a measure of blastogenesis. The responses to Con A of all infected calves were significantly depressed while the responses to PHA were not. SLA, at concentrations of 4 micrograms ml-1 and above, caused blastogenic activity in lymphocytes from uninfected calves. Using SLA at 1 microgram ml-1 in lymphocyte cultures supplemented with autologous serum, an antigen-induced blastogenic response was detected in calves receiving serial inoculations of L3.  相似文献   

16.
An in vitro immunization procedure was developed, whereby bovine blood and mammary gland lymphocytes were isolated, and their blastogenic responses (measured by [3H]thymidine uptake) to sensitization and subsequent stimulation with selected antigens were determined. Capsular extracts of Staphylococcus aureus, encapsulated S aureus, and nonencapsulated S aureus were used as test antigens. Differences were observed between kinetics of the secondary response to encapsulated and nonencapsulated S aureus. The secondary response to nonencapsulated S aureus peaked in 48 hours, whereas the secondary response to the encapsulated S aureus peaked in 72 hours. The delayed peak response to encapsulated S aureus was observed only when encapsulated S aureus was used for sensitization of lymphocytes, regardless of whether the encapsulated or nonencapsulated strain was used for stimulation. Sensitization of lymphocytes with the nonencapsulated strain and stimulation with the encapsulated strain did not alter the kinetics of the secondary response of cells sensitized with the nonencapsulated strain. Seemingly, T and B cells were responsive to in vitro immunization with S aureus antigens.  相似文献   

17.
Levamisole and its influence on the immune response of lambs   总被引：1，自引：0，他引：1  
Ten parasite-free lambs were drenched with 8 mg/kg of levamisole on days 0 and 28 and were injected with human erythrocytes and ovalbumin one day after each drench. Ten other antigen-injected lambs were not drenched with anthelmintic as controls. Lymphocytes from the control and drenched lambs were culturedin vitro with RPMI 1640 plus 5% fetal calf serum (FCS), with 50% autologous serum only, with concanavalin A (Con A) or with phytohaemagglutinin (PHA). Decreased blastogenesis was observed in cells from the drenched lambs cultured in the presence or absence of mitogen and was most obvious when 50% autologous serum was used, particularly with PHA, and when lymphocytes were collected 3 and 7 days after the first and 3 days after the second antigen injection. There were no significant changes in antibody titres between the groups. Decreased serum complement activity was seen 3 days after the second antigen injection in the drenched lambs. Although there was a significant reduction in the serum insulin-like growth factor I levels 4 days after each levamisole drench, the drenched lambs gained significantly more weight than the non-drenched control lambs.Abbreviations Con A concanavalin - EIA enzyme immunoassay - FCS fetal calf serum - GH growth hormone - IGF-I insulin growth factor I - PHA phytohaemagglutinin  相似文献   

18.
The Jerne hemolytic plaque assay was used to compare the number of antibody forming cells in the ipsilateral supramammary/suprainguinal lymph node which drains the udder, its counterpart area in males, of dairy goats inoculated with the antigen, sheep red blood cells, and in the contralateral lymph node which drains the corresponding non-inoculated area. Parenteral immunization was shown to have suppressing effects upon the local immune responses to the subsequently applied antigens. Three monthly intramammary inoculations of the antigen induced significant numbers of indirect plaque-forming cells (i.e. immunoglobulin G antibody producing cells) in both draining (ipsilateral) and non-draining (contralateral) nodes, suggesting antigen relocation and/or cell relocation from the inoculated side. However, the number of indirect plaque-forming cells in the ipsilateral node was far greater than that in the contralateral node, indicating that the majority of memory cells remained in the inoculated site.  相似文献   

19.
五种旋毛虫抗原对猪的免疫保护作用研究   总被引：4，自引：0，他引：4  
本实验研究了旋毛虫肌幼虫可溶性粗抗原、排泄分泌抗原（ＥＳ）、表面抗原（ＳＡ）及成虫ＥＳ、ＳＡ５种抗原对猪的免疫保护作用。结果５种抗原对猪均具有一定程度的免疫原性，可诱导猪体产生对攻击感染的抵抗力（减虫率），其中肌幼虫粗抗原为５５．２０％；肌幼虫ＥＳ为４２．５６％，肌幼虫ＳＡ为７２．２１％；成虫ＥＳ为３２．９２％；成虫ＳＡ为４２．１７％。免疫５种抗原后用肌幼虫“Ｂ”抗原、新生幼虫可溶性抗原及成虫可溶性抗原进行ＥＬＩＳＡ检测，均可测出血清抗体应答反应，其中以相应抗原测出的抗体应答较强烈。免疫５种抗原后猪外周血液中Ｂ淋巴细胞减少，Ｔｈ及Ｔｓ增加，Ｔｈ／Ｔｓ比值降低，呈暂时的细胞免疫抑制现象。  相似文献   

20.
Fourteen sheep persistently infected with border disease virus were investigated to examine the effects of persistent viraemia on lymphocyte subpopulations in peripheral blood and their responses to the mitogen phytohaemagglutinin (PHA) in vitro. Persistently infected sheep had significantly more CD8+ (cytotoxic-suppressor) T-lymphocytes than uninfected sheep of the same age (P less than 0.001). The total number of CD4+ (helper) T-lymphocytes were not significantly different but there were more T-lymphocytes (CD5+) which were CD4- and CD8- in normal sheep than persistently infected sheep (P less than 0.001). Peripheral lymphocytes obtained from persistently infected sheep showed significantly reduced blastogenesis induced by PHA than those obtained from normal sheep (P less than 0.001).  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号