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1.
Cattle exposed to their third consecutive warble (Hypoderma lineatum and H. bovis) infestation had significantly reduced apparent and accumulative grub populations and produced significantly fewer grubs than animals exposed to their first infestation. These resistant animals had enhanced reactivity to mitogens (concanavalin A (con A) and pokeweed) upon reinfestation and a strong antigen-specific response at 2 months post-infestation. Their responsiveness to con A and antigen was also enhanced at 7 months post-infestation when grubs appeared in the backs, and the response to antigen continued at 10 months post-infestation. Responsiveness to mitogens in previously uninfested animals was similar to the control response throughout the infestation apart from the suppression of the con A response at 7 months post-infestation. These animals finally responded to antigen at 10 months post-infestation. These results suggest that acquired resistance to hypodermosis has a cellular basis with participation of both B- and T-cell components.  相似文献   

2.
The influence of an antigen-specific cellular and humoral immune response, stimulated by immunization, on survival of a challenge infestation of Hypoderma lineatum was investigated. Calves immunized with a purified combination of hypodermin A, B and C plus monophosphoryl lipid A (MPL) developed a strong antigen-specific cellular immune response by completion of the immunization schedule which persisted to 12 weeks post-infestation. Responsiveness of peripheral blood lymphocytes to the mitogens concanavalin A and pokeweed was also elevated at 4 and 12 weeks post-infestation. Western blot analysis at the time of maximum grub counts demonstrated that immunized calves responded to hypodermin A, B and C while those receiving only MPL or infested controls responded only to hypodermin B and C. The antigen-specific antibody response as measured by ELISA at maximum grub count was significantly higher in vaccinated calves than in infested controls while the response in calves receiving only immunostimulator was also significantly elevated. Immunized (antigen plus MPL) calves produced 5.0 +/- 6.9 grubs per animal which successfully pupated while those receiving MPL alone produced 16.4 +/- 6.1 and infested controls produced 32.2 +/- 10.9 grubs per animal.  相似文献   

3.
The understanding and importance of antigen-specific immune responses after vaccination has completely changed in recent years. In the past, the focus for monitoring a vaccine-specific immune reaction was principally on the humoral branch of the immune system. The efficacy of vaccines, as assessed by the induction of protective immunity was mainly correlated with antibodies and antibody-titers. However, this correlation often failed and other parts of the immune system had also to be considered: namely, the innate immune system and the cellular branch of the antigen-specific immune system. With regard to vaccines, the innate immune system plays its main role in the effective activation of the antigen-specific immune response, in antigen-uptake and antigen-presentation. The dendritic cells (DCs) are the most important antigen presenting cells which present processed protein antigens (peptides) through MHC-molecules: MHC-class I, for the presentation of endogenous synthesised antigen; MHC-class II for exogenous antigen. Activation of DC leads to an enhanced production of cytokines and chemokines, to an up-regulation of co-stimulatory and activation molecules and also molecules for cell-cell interactions, e.g. interactions with cells of the antigen-specific immune system. T lymphocytes are the effector cells of the cellular branch of the antigen-specific immune system. They act either as MHC-class I-restricted cytolytic T lymphocytes (CTL) or as MHC-class II-restricted T-helper cells providing support for B lymphocytes (T(H)2) and the cellular part of the antigen-specific immune system (T(H)1). In order to achieve effective vaccination, the activation of all T-cell subpopulations is of advantage, but more important is the generation of antigen-specific memory T and B lymphocytes. In addition to these 'generic' immunological factors which are essential for the design of more efficacious vaccines, our detailed knowledge about feline and canine immune reactions after vaccination, which is still poor, has to be improved.  相似文献   

4.
Lymphocytic responses in peripheral blood and visceral lymph to Cooperia oncophora antigen and skin tests were determined in 35 Holstein male calves that were inoculated orally with single or multiple doses of C oncophora infective larvae. Several calves were vaccinated or given immune serum before larvae were inoculated. Antigen-specific in vitro blastogenesis of blood and lymph lymphocytes and delayed-type hypersensitivity reactions were observed in several inoculated, vaccinated, and/or passively immunized calves. Most calves that had delayed skin reactions also had in vitro lymphocyte responses to C oncophora antigen. The lymphocyte and skin responses were inconsistent and variable in time of onset--the earliest lymphocyte response occurring 7 days after calves were inoculated. A cellular immune response was induced by both dermal vaccination and oral inoculation; however, passive immunization by IV administration of immune serum simultaneously with inoculation did not have an apparent effect on the cellular response, as measured by the lymphocyte blastogenesis test or dermal testing. Although cellular immune responses were observed in several calves infected with C oncophora, there was no apparent relationship between the specific responses and number of nematodes establishing infection in calves after either single- or multiple-dose oral inoculations.  相似文献   

5.
The potential for cross-transmission of Hypoderma lineatum from cattle to domestic goats (Capra hircus) was examined using artificial infestation techniques. Two routes of infestation, subcutaneous injection and dermal penetration, were used to expose goats to newly hatched first instars. Presence of antibodies and appearance of circulating antigen (hypodermin C) were evaluated at selected intervals for up to 40 weeks post-infestation. In addition, immunoblots against H. lineatum first-instar proteins were conducted using sera taken at 10 weeks post-infestation. Goats were palpated for the presence of developing larvae at sub-dermal sites beginning at week 30 pi. No developing larvae were palpated at any time, regardless of the route of infestation nor was circulating antigen detected in any infested goats. Antibodies were present at weeks 6 and 10 and week 27 pi in both infested groups. Immunoblots indicated all infested goats produced antibodies to first instar H. lineatum antigens. H. lineatum appears to be incapable of completing development in domestic goats although the transient appearance of ELISA detectable antibodies and the presence of bands on immunoblots suggests that at least some larvae survive long-enough to engender a humoural response. The host specificity of H. lineatum is discussed in light of the general concepts of host-parasite relationships of oestrids.  相似文献   

6.
The collagenase (hypodermin C) from soluble crude extracts of Hypoderma lineatum 1st-instar larvae was purified by reverse-phase HPLC and used in a new indirect ELISA test. This pure protein had several advantages over the use of crude larval extracts allowing a much better discrimination between infested and non-infested cattle. The anti-hypodermin C titers of 19 Asturiana cattle were estimated over the course of a natural H. lineatum infestation cycle, in which the effect of ivermectin treatment was also investigated. The results showed differences in the onset and ending of the infestation with respect to those described for other European countries. The ivermectin treatment proved to be very effective and treated animals had relatively low anticollagenase titers.  相似文献   

7.
Responses of infested and vaccinated Hereford cattle to Boophilus microplus antigens were measured by enzyme-linked immunosorbent assay (ELISA), lymphocyte blastogenesis assay (LBA) and intradermal skin tests. Responses against soluble salivary gland extracts (SGS), salivary gland membrane (SGM), soluble gut extracts (GS), gut membrane (GM), soluble larval extracts (LS) and larval membrane (LM) antigens were tested. In one experiment, cattle infested with up to 160,000 ticks had positive cellular responses to SGS and significant antibodies against LM, GM, SGM, and SGS. Cellular responses to Concanavalin A were not depressed following infestation. Cattle vaccinated with GM, using Quil A as adjuvant, had positive cellular responses to gut and salivary gland antigens and significant antibody responses to all antigens tested. The antibody levels of vaccinated cattle were significantly higher than the antibody levels of infested cattle (P less than 0.05). In a second experiment, immune responses of cattle infested with 40,000 ticks were studied during 38 days. Cellular responses in LBA to several tick antigens were transiently elevated and significant levels of antibody were measured against LM, GM, SGM and SGS, from day 25 (P less than 0.05). Infested cattle had positive skin reactions following intradermal injection of larval and adult tick antigens (P less than 0.05).  相似文献   

8.
Oesophagostomum radiatum, the nodular worm of cattle, is a severe pathogen in previously uninfected calves. However, cattle develop a strong protective immune response upon exposure to the parasite. In order to evaluate whether soluble parasite antigens could induce protective immunity, a soluble fraction was obtained from disrupted adult worms, and this fraction was used to vaccinate calves. The vaccination protocol involved two immunizations. The first was administered intramuscularly with complete Freund's adjuvant, the second was given intraperitoneally with antigen plus alum. This immunization reduced the number of worms developing from a subsequent challenge infection by 85% and also reduced clinical signs associated with infection with adult worms. However, vaccination resulted in decreased weight gains during the larval phase of the infection. Analysis of the immune response generated in the vaccinated calves indicated that protection from infection was significantly correlated with the levels of: (1) circulating parasite-specific IgG2 antibody; (2) cellular immune reactivity as determined in a conventional parasite-specific lymphocyte proliferation assay. Serum anti-O. radiatum IgG2 antibodies from vaccinated calves were used in immunoblots to identify the major immunogens. There were five major immunogens with molecular weights ranging from 70 to 150 kDa. Fractions separated by high-pressure liquid chromatography contained immunogens that were used to immunize calves. Vaccination with these fractions was found to impart the same level of protective immunity and induced similar IgG2 antibody and cellular immune responses as the crude whole worm extract even with 100-fold less protein.  相似文献   

9.
Stage specific survival of larvae and pupae of the common cattle grub, Hypoderma lineatum de Villers, was compared in cattle artificially infested with larvae delivered by subcutaneous injection as a single infestation of 100 larvae, in trickle infestations of either two doses of 50 larvae, separated by 15 days, or in four doses of 25 larvae, separated by 9, 6, and 6 days, respectively. The duration of the migratory and 'warble' phases were also compared between treatments. The kinetics of the antibody response was monitored throughout the larval development by ELISA. No difference was noted in the level of antibodies among the three treatments. Mortality of first-instars differed among the infestation groups with mortality in the single pulse infestation group相似文献   

10.
An antigen capture or sandwich ELISA (sELISA) was evaluated for the diagnosis of Hypoderma lineatum in cattle under field conditions in northwestern Spain. The kinetics of circulating hypodermin C (HyC) and specific antibodies during the course of natural infestation were determined in a group of 10 Frisian calves. In addition, oesophagi and blood samples were taken from 105 cows at a slaughterhouse in order to compare three methods for the diagnosis of H. lineatum: sandwich ELISA for the detection of the antigen HyC (sELISA), indirect ELISA for the detection of antibodies anti-HyC (iELISA) and the detection of first instars (L1) in the oesophagus. In naturally infested cattle, HyC was present in circulation at low levels during the early and late phases of the infestation. However, in the middle phase, coinciding with the presence of L1 in the oesophagus, two peaks of increased HyC concentration were observed. Specific antibodies increased progressively until the first appearance of larvae in warbles on the back. There was no correlation between antigen or antibody levels and the number of grubs in the back. Prevalence of first instars in the oesophagi of slaughtered cows was 21.9% (23/105). The percentage of cattle that were positive for circulating antigen was slightly higher (24.8%), suggesting the recent destruction of migrating larvae in some animals. However, there was no correlation between the number of L1 and HyC levels. With the iELISA, 79% of the animals were positive to Hypoderma, which means that a high percentage of those animals have been exposed to the parasite but they had no apparent current infestation. The sELISA is a good tool to follow larval development within the host; however, the episodic elevation of HyC levels limits the usefulness of this test for the early diagnosis of Hypoderma under field conditions.  相似文献   

11.
West Nile virus (WNV) is a single-stranded, enveloped RNA virus capable of causing encephalitic disease in horses. Unvaccinated horses are at risk for developing WNV disease in endemic geographic regions. Effective vaccination reduces disease frequency and diminishes disease severity in vaccinated individuals that become infected with WNV. Recent data indicate CD4+ lymphocytes are required for effective protection against disease; in particular, cross talk between CD4+ and CD8+ lymphocytes must be functional. The objective of this project was to investigate immune responses in horses throughout a series of three vaccinations using a commercial inactivated vaccine under natural conditions. Immune responses to vaccination were determined by neutralizing antibody titers with plaque reduction neutralization test (PRNT), IgM titer (capture ELISA), WNV specific antibody Ig subclass responses, WNV lymphocyte proliferative responses and intracellular cytokine expression. Horses were vaccinated with a series of three vaccines at 3-week intervals using an inactivated product. An initial measure of immune activation following vaccination was determined by evaluating changes in lymphocyte cytokine expression. Interferon (IFN) gamma and interleukin (IL)-4 expressing CD4+ lymphocytes significantly increased 14 days following initial vaccination compared to unvaccinated horses (P<0.05). IFN-gamma expressing CD8+ lymphocytes also increased and remained elevated for 110 days. Antigen specific lymphocyte proliferative responses were significantly increased up to 90 days following the third vaccination (P<0.05). As expected, vaccinated horses produced increased neutralizing antibody based on PRNT data and WNV antigen-specific Ig subclass responses compared with unvaccinated horses (P<0.05). Our data indicate that WNV vaccination with an inactivated product effectively induced an antigen-specific antibody responses, as well as CD4+ and CD8+ lymphocyte activation.  相似文献   

12.
Serpulina hyodysenteriae infection of pigs, swine dysentery, causes a mucohemorrhagic diarrhoea resulting in significant economic losses to swine producers. The pathogenesis of this disease is poorly understood. Regardless, commercial vaccines have been developed and are in use. Thus, the present study was designed to examine cellular immune responses induced by parenteral S. hyodysenteriae vaccination. Significant antigen-specific interferon-gamma (IFN-gamma) and blastogenic responses were detected from peripheral blood lymphocytes isolated from vaccinated pigs. However, poor IFN-gamma responses were detected from colonic lymph node lymphocytes from these same pigs despite significant antigen-specific blastogenic responses. In addition, peripheral blood IFN-gamma responses were diminished by either in vitro depletion of CD4 expressing cells or by in vitro treatment with porcine IL-10. Colonic lymph node IFN-gamma responses were not inhibited by treatment with porcine IL-10. Vaccination also resulted in increased percentages of both mucosal and peripheral blood CD8 single positive cells with concurrent decreases in percentages of CD4 single positive cells as compared to percentages of these same populations from non-vaccinated pigs. In conclusion, these studies show that parenteral S. hyodysenteriae vaccination results in cellular immune responses detectable both peripherally (systemic immunity) as well as at the site of infection (mucosal immunity). However, it appears that regulatory mechanisms affecting IFN-gamma production in response to S. hyodysenteriae antigen differ between peripheral and colonic compartments.  相似文献   

13.
The feasibility of immunizing calves against Hypoderma lineatum and H. bovis with a crude H. lineatum larval extract or culture-derived antigen was investigated. Larval survival was followed through pupation. Grub appearance in the backs of both of the immunized groups was found to be 50% of that in the control groups. First appearance of H. bovis was significantly retarded in both immunized groups; however, H. lineatum was unaffected. Duration of H. bovis dropping was also shorter in animals receiving crude extract than in controls while duration of dropping of H. lineatum was not influenced by immunization. Survival of H. lineatum larvae to pupation was significantly reduced in animals immunized with crude larval extract, but not in those receiving culture-derived antigen. H. bovis survival was reduced by both treatments.  相似文献   

14.
The development of improved vaccines for bovine tuberculosis is urgently required as a cost effective solution for control and eventual eradication of tuberculosis in domestic animals. Studies in small animal models of tuberculosis have shown that vaccination with culture filtrate proteins (CFP), prepared from Mycobacterium tuberculosis or M. bovis, can induce cellular immune responses and confer a level of protection against aerogenic challenge with virulent mycobacteria. As a first step in the development of a mycobacterial CFP vaccine for protection of cattle against bovine tuberculosis, the immune responses of cattle vaccinated with short-term culture filtrate proteins (ST-CFP) from M. tuberculosis and formulated with different adjuvants were compared with those vaccinated with bacille Calmette-Guerin (BCG). The adjuvants included dimethyldioctyldecyl ammonium bromide (DDA), diethylaminoethyl (DEAE)-dextran, and ST-CFP adsorbed onto polystyrene beads. Vaccination with ST-CFP/DEAE-dextran induced high levels of interleukin-2 (IL-2) but low levels of interferon-gamma (IFN-gamma) from whole-blood cultures stimulated with M. tuberculosis ST-CFP in comparison with the strong IFN-gamma and IL-2 responses induced after vaccination with BCG. ST-CFP/DEAE-dextran also induced a strong antigen-specific immunoglobulin antibody response with both immunoglobulin G1 (IgG1) and IgG2 isotypes. Vaccination with ST-CFP/beads induced a weak IgG1-biased antibody response but no IFN-gamma or IL-2 response. DDA did not induce significant immune responses in animals vaccinated with ST-CFP. In comparison to the moderate delayed-type hypersensitivity (DTH) responses induced by vaccination with subcutaneous BCG, none of the ST-CFP vaccines induced a significant DTH response to either M. tuberculosis ST-CFP or bovine purified protein derivative (PPD). While the ST-CFP vaccines used in this study have not induced strong antigen-specific cellular immune responses in cattle comparable to those induced by BCG, they are immunogenic in cattle and it may be possible to overcome this problem by using adjuvants that more effectively promote IFN-gamma responses in this species.  相似文献   

15.
To better understand the interaction between Mycoplasma bovis and its bovine host, we have characterized the immune response generated during an experimental lung infection with M. bovis. Proliferation ([3H]-thymidine blastogenesis) and Th1/Th2 cytokine production were used to monitor peripheral cellular immune responses. Flow cytometry analysis was used to determine T-cell subset activity by CD25 expression. Humoral immune response was monitored by the identification of antigen-specific IgG1 and IgG2 isotypes over time. Herein, we show that M. bovis antigen stimulates activation of CD4+ and CD8+ cells in vitro in a manner consistent with memory, and that gammadelta-T cells are activated by antigen in a manner consistent with innate immunity. In addition, the percentage of cells producing IFN-gamma during recall response is equal to that of IL-4 producing cells. Serological analysis shows M. bovis stimulates increased production of antigen-specific IgG1 while very little IgG2 is produced. We therefore submit that experimental lung infection of cattle with M. bovis results in a Th2-skewed immune response.  相似文献   

16.
Between 1989 and 1997 different factors related to warble fly infestation (prevalence, intensity, climate, breed) were studied in north-eastern Algeria. Mean prevalence during the period was 76%. This figure was obtained from a cattle market survey of four different regions (Jijel, Constantine, Guelma, and El Tarf). One-hundred percent of the herds were found to be infested in El Tarf and 61% of animals were infested. Within a herd, the intensity of infestation decreased with the age of the cattle. The mean number of warbles per animal decreased as reinfestations occurred. The peak of the emergence of warbles observed between January and March was progressively delayed in the older animals. Two species of Hypoderma were present in cattle in Algeria (H. bovis and H. lineatum) with a predominance of H. lineatum (63%) in the semi-arid area (Batna) and a majority of H. bovis (75%) in humid area (Jijel). Warbles appeared earlier in Batna (October) than in Jijel (November). The Prim'Holstein breed was more susceptible to the warble fly infestation than Montbeliard or local breeds. The implementation of a control programme for hypodermosis in Algeria is urgent. Control can be achieved through a mass treatment of cattle. This approach must take into account the factors described in this study which influence the epidemiology of the disease.  相似文献   

17.
为了解动物在注射口蹄疫灭活疫苗后机体免疫状态的变化情况,本试验选取口蹄疫阴性猪进行疫苗免疫,利用流式细胞术检测方法来测定外周血中免疫细胞亚群的变化。结果显示,树突状细胞在免疫后能够快速产生应答,在24 h内其含量出现明显升高;B细胞和Th细胞含量在免疫后第10天都出现了明显升高,且两者之间的变化存在着高度相关性,而抗体水平的变化情况与这两种细胞含量的变化也高度相关;细胞毒性T细胞含量在免疫后也出现升高。这说明灭活疫苗免疫动物后不仅可以激活机体的体液免疫应答反应,产生高水平抗体,发挥免疫保护作用,更重要的是能够在短时间内激发机体的固有免疫应答,同时能够激活细胞免疫应答,全方位发挥免疫保护作用。  相似文献   

18.
Identification of an immune response correlate for protection against bovine tuberculosis would greatly facilitate the rational development of an effective vaccine. However, finding such a correlate has been a daunting task. Vaccination/challenge studies in cattle provide an ideal platform to compare induction of immune response parameters following vaccination and challenge, and assess the correlation of these parameters with protection. Protection against tuberculosis requires a Th 1-type cell-mediated immune response and induction of an antigen-specific interferon-gamma (IFN-gamma) response was the logical first choice in an investigation to identify an immune response correlate for protection. Calf vaccination studies showed that the subcutaneous injection of BCG vaccine induced significant protection against experimental challenge with Mycobacterium bovis. This protection was associated with strong whole blood IFN-gamma responses to bovine PPD 2-4 weeks after vaccination, but within the BCG-vaccinated groups, these responses were not correlated with protection. Use of a variety of vaccination strategies has shown that IFN-gamma responses in isolation were not necessarily associated with protection and concurrent IL-4 mRNA expression or antibody responses could also be induced. Collation of an immunological profile may be more informative than a study of individual cytokines. An indication of vaccine efficacy can be provided by the study of immune responses following challenge of the calves with M. bovis. IFN-gamma responses to ESAT-6, antibody responses following tuberculin skin testing and antigen-specific IL-4 mRNA expression all correlated with the severity of disease and indirectly provided an indication of protection. Future studies should be directed towards obtaining immunological profiles of calves following vaccination using techniques such as DNA microarray analysis, measurement of cytokine mRNA expression by real-time PCR, protein profiling by SELDI-TOF mass spectrometry as well as determining cytokine production by specific T cell sub-sets in individual protected animals.  相似文献   

19.
In order to investigate the change of lymphocyte subpopulations in the peripheral blood after injection of foot-and-mouth disease inactivated vaccine, blood samples were collected after vaccination and assayed with flow cytometry.The results showed that the proportion of dendritic cells (DCs) increased significantly in a short time after vaccination, after which the number of B cells, helper T (Th) cells and cytotoxic lymphocytes (CTLs) also raised with significance.The changing trend of B cells proportion was highly relevant with Th cells and a same correlation also appeared in the antibody titers.This indicated that the innate immune system could be activated rapidly after vaccination with the inactivated vaccine, and the adaptive immune response including cellular immune response and humoral immune response could be invoked subsequently.The high level of neutralizing antibodies was of great importance in evading FMDV and the cellular immune response also exerted a crucial role.  相似文献   

20.
Purebred Holstein calves acquired resistance to Amblyomma americanum adult ticks after 1 infestation and expressed resistance during a challenge infestation 26 days later. Tick yields from resistant animals were normal, but mean tick weight and egg-mass weight were reduced significantly (31 and 32%, respectively). Cutaneous cellular responses at tick feeding sites in primary (naive) hosts consisted almost exclusively of mononuclear cells, until 24-h post-infestation when granulocytes appeared. At 48 h, basophils were the most abundant granulocyte and comprised 19% of the total cellular response. Secondary and tertiary hosts expressed significant cutaneous basophil responses as early as 6 h post-tick infestation, indicative of an anamnestic response. Cutaneous basophil levels in tertiary hosts (16-34% of the infiltrate) were increased by 2-fold over secondary hosts (12-16% of the infiltrate). Neutrophils and eosinophils were fairly abundant in secondary hosts (22-33%, and 3-13%, respectively), but decreased significantly in tertiary hosts corresponding to increased basophil levels. Mast cells comprised less than 10% of the cells found in primary hosts, and less than 5% in secondary and tertiary hosts representing a consistent, but insignificant component of the total cells. Basophils were not observed in the blood of hosts during any tick infestation, but a peripheral blood basophilia (0.1-1.1%) marked the end of each tick feeding period. A neutropenia was observed in the blood of tertiary hosts corresponding to a decreased tissue neutrophil response. Blood eosinophil, lymphocyte and monocyte levels remained unchanged during each tick infestation compared to controls. In summary, calves acquired immunity to A. americanum ticks, resulting in significant decreases in feeding and ovipositional success associated with a local cutaneous basophil response and peripheral blood basophilia. These findings suggest that basophils are involved in bovine immunity to ticks as has been established in the guinea pig model.  相似文献   

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