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1.
Neutrophils, eosinophils and macrophages interact with invading parasites and naive hosts. The initial reaction of leukocytes is the generation of reactive oxygen species (ROS). The cytotoxic effects of extracts derived from intact Cysticercus cellulosae and from the scolex or membrane fractions on neutrophils were examined. DNA fragmentation of neutrophils was observed when cells were incubated with an extract from the intact metacestode; however, the addition of antioxidant enzymes to the incubation medium had a protective effect. The scolex and membrane extracts did not affect DNA fragmentation of neutrophils. Hydrogen peroxide production of neutrophils incubated with metacestode fractions from C. cellulosae increased by 190% (total extract), 120% (scolex) or 44% (membrane). An increase in antioxidant catalase activity (28%) concomitant with the increased production of ROS was observed in neutrophils incubated with metacestode fractions, which could be an attempt at self-protection. ROS production by neutrophils in the presence of the intact cysticerci extract did not alter phagocytosis. In contrast, the scolex and membrane fractions increased the phagocytic capacity of neutrophils by 44 and 28%, respectively. The results showed that the extract from intact C. cellulosae was toxic for neutrophils via ROS production, leading to DNA fragmentation and inhibition of phagocytic capacity, but neutrophils are able to protect themselves against oxidative stress by via catalase activity.  相似文献   

2.
采用Sephadex G-200层析技术纯化猪囊尾蚴头节抗原,用纯化抗原包被ELISA板,建立间接ELISA方法。通过各种条件优化,最终确定最佳试验条件为:抗原最适包被量为30mg/L;血清稀释度为1∶800;血清最佳反应时间为60min。结果表明,该方法具有较好的稳定性和重复性,可用于分泌抗猪囊尾蚴头节单克隆抗体杂交瘤细胞株的筛选。  相似文献   

3.
猪囊尾蚴抗原成分及其诊断价值分析   总被引:3,自引:0,他引:3  
猪囊尾蚴病是一种重要的人畜共患寄生虫病。囊尾蚴病的快速诊断是囊尾蚴病治疗和预防的前提,免疫学诊断方法是近年来的囊尾蚴病临床诊断中的重要辅助手段,高特异性诊断抗原的筛选为近年来的研究热点。囊尾蚴抗原组成复杂,包括囊液抗原、头节抗原、囊壁抗原、循环抗原和排泄分泌抗原等。其中的囊液抗原特异性较好,可用于免疫学诊断,而排泄分泌抗原诊断囊尾蚴的价值尚待进一步研究。  相似文献   

4.
应用PAGE-IEF分析了三种绦虫的可溶性蛋白质。猪囊尾蚴头节囊壁、囊液,细颈囊尾蚴头节囊壁、囊液和曼氏迭宫绦虫成虫蛋白质区带数分别为30、25、29、18和24~29条,主带(峰)数分别为10、9、9、7和12个,面积最大峰分别为J、B、H、F和E峰。五种样品电泳谱在总带数、各pH区带数、主带(峰)数和分布、面积最大峰都明显不同,可供鉴定和鉴别。细颈囊尾蚴蛋白质电泳分析与猪囊尾蚴的比较分析,以及曼氏迭宫绦虫蛋白质电泳分析系国内外首次报道。  相似文献   

5.
本研究测定了体内发育过程中猪囊尾蚴钠钾三磷酸腺苷酸(Na^ K^ -ATPase)、钙三磷酸腺苷酸(Ca^ -ATPase)和镁三磷酸腺苷酸(Mg^ -ATPase)活性变化,结果表明囊壁部的Na^ K^ -ATPase和头颈部Ca^ -ATPase活性在体内发育60、80、95天高于或显著高于体内发育30、40天。提示随虫体生长发育相应代谢通路加强。  相似文献   

6.
The sensitivity and specificity of the enzyme-linked immunosorbent assay (ELISA) for the diagnosis of Taenia solium cysticercosis was evaluated in experimentally and naturally infected pigs, using T. solium larval scoleces and its fractionated 1st and 2nd peaks on Sephadex G-200 as antigens. First peak antigen gave maximum sensitivity and highest antibody titres. The overall sensitivity of this test was found to be 91.5, 95.8 and 70.8% with scolex, 1st and 2nd peak antigens, respectively. False positive reactions occurred in 9.09% of uninfected pigs with scolex and 1st peak antigens and cross-reactions occurred in 25% of Taenia hydatigena-infected animals using scolex and 2nd peak antigens. No cross-reaction was observed using 1st peak antigen. The specificity of the test was 92.3, 96.2 and 92.3% with scolex, 1st and 2nd peak antigens, respectively.  相似文献   

7.
Crude larval Taenia solium extracts were fractionated by Sephacryl S-200 gel filtration into four fractions (W1-W4). The sensitivities of the fractions to rabbit and pig antiserum against Taenia solium were tested by double immunodiffusion, immunoelectrophoresis, and ELISA. Fraction W2 which was highly sensitive to antisera was shown by immunoblotting to contain antigen B (95 and 105 kDa). The four fractions were shown to contain antigenic determinants common with pig serum proteins and crude extracts of other Platyhelminthes (especially Taenia hydatigena). Fraction W2 has the potential to be used as a serodiagnostic antigen.  相似文献   

8.
建立了8株抗猪囊虫循环抗原(CA)的杂交病细胞系。其中6F12和2E7为抗囊虫特异McAb;McAb 1C6、1C7、1B5、4D9、2B9和8D8与囊虫、(?)球蚴、细颈囊尾蚴抗原均可发生反应。这些McAb的腹水ELISA效价为105~107,细胞培养上清液效价为102,并均可与病猪血清中的囊虫CA反应形成沉淀线。用1B5、6F12和8D8分别致敏血球,以反向间接血凝试验检测98份囊虫病猪血清,检出率分别为70.41%(69/98)、6.12%(6/98)和7.14%(7/98)。本研究制备的McAb可用于猪囊虫循环抗原检测。  相似文献   

9.
A protective antigen was purified from a saline extract of a Type 1 strain of Pasteurella multocida by chromatographic methods, and its chemical and immunological ccharacteristics were studied. Three protein peaks were obtained from crude extract by gel filtration with Sephadex G-200. A bacteria-specific antigen was detected only in the first peak fraction, which, after passing through an immunoadsorbent column to remove any components originating from the growth medium, was adsorbed onto DEAE-cellulose followed by elution with a gradient of NaCl. From the first peak fraction of the gel filtration, 4 protein peaks were obtained, the second and third peaks being the major ones. Carbohydrate/protein ratios of the peak fractions varied from 0.06 to 1.0. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that 2 proteins of molecular weights 44 000 and 25 000 were present in all the fractions. The 4 DEAE-cellulose fractions (DP-1 to DP-4) contained a single antigenically identical material, and induced protective immunity in turkeys against challenge exposure. The second peak fraction from DEAE-cellulose (DP-2) protected turkeys when subcutaneously injected as 2 doses of 10 μg protein with a 14-day interval between doses. The DP-2 fraction induced antibodies in rabbits which formed a single precipitin line against the crude extract. The purified antigen (DP-2) from a Type 1 strain was antigenically distinct from a similar antigen purified from a Type 3 strain; there was no significant cross protection in turkeys between the 2 antigens. These results indicate that protective antigens purified from soluble extracts of a Type 1 or Type 3 strain possess similar physicochemical properties, but that they are immunologically distinct from each other.  相似文献   

10.
The present study was carried out to identify the excretory/secretory (E/S) antigens of the rumen infecting digenetic trematode Gastrothylax crumenifer that may be useful for the immunodiagnosis of rumen amphistomosis particularly during the pre-monsoon season during which this rumen parasite stops shedding eggs. The in vitro released E/S proteins were purified on a Sephadex G-200 column. The gel filtration profile revealed three distinct fractions F1-F3 where F1 and F3 appeared as sharp peaks while the F2 fraction was dispersed. The antibody titre against each of the purified E/S fractions was determined by ELISA using anti-whole E/S polyclonal antibodies raised in rabbit. Among the three fractions, the antibody titre against F1 was highest (1:12,800) whereas IgG titre was very low (1:50) for fraction F2 and F3 (1:100). Of the total polypeptides resolved on gradient SDS-PAGE, only a few antigenic polypeptides were detected in each fraction with hyperimmune anti-serum as revealed by Western Blot analysis. However, a 33 kDa antigen detected in each fraction appeared to be immunodominant which could be exploited for the diagnosis of the pouched amphistome.  相似文献   

11.
The antigenic mosaics of three Bacteroides nodosus isolates (198, 199 and 127) were studied to elucidate the nature of the protective immunogen. In vaccinated sheep the three isolates induced high homologous serum agglutinin titres but it was also apparent that 198 and 199 shared a major surface antigen not present on 217. This major cross-reacting antigen was not detected with rabbit antisera. The fimbriae, consisting predominantly of protein, induced high homologous titres in rabbits and represented the type-specific antigen. Lipopolysaccharide (LPS) from each of the isolates induced low agglutinin titres and high 2-mercaptoethanol-sensitive indirect haemagglutinating antibody titres. The heat-stable LPS contained at least two common carbohydrate O antigen determinants but no type-specific O antigens were detected.  相似文献   

12.
应用琼脂双扩散(DID),免疫电泳(IEP),聚丙烯酰胺凝胶电泳(PAGE)对猪囊尾蚴的匀浆抗原和囊液抗原进行分析。结果表明,两种抗原间存在相同的抗原成份,囊液的免疫化学性质优于匀浆抗原,并从免疫学和生物化学角度初步探索了囊液抗原和匀浆抗原中蛋白质之间的相互关系及主要蛋白质组分。  相似文献   

13.
Examination by immuno-diffusion methods in agar gel plates demonstrated that the supplementing fraction precipitated from normal bovine serum by dialysis against dilute buffer, pH 5.0 to 6.6, contains at least three main antigens, and a weaker fourth. Two of these antigens or antigen determinants appeared to be present in globulin fractions prepared by dilution with distilled water or differential precipitation with ammonium or sodium sulphate.  相似文献   

14.
Six commercially available clostridial vaccines comprising one oil-emulsion, two alum-precipitated and three aluminum hydroxide adjuvanted preparations, each containing between two and seven antigenic components, were administered to groups of 10 rabbits and eight sheep in accordance with manufacturers' recommendations. Serum antitoxic values to Cl welchii beta, Cl welchii epsilon, Cl septicum, Cl oedematins and Cl tetani toxins were determined 14 days after completion of each vaccination course. The overall pattern of mean antitoxic values was found to be similar in sheep and rabbits, a vaccine eliciting a comparatively high antibody titre to any given antigen component in sheep also inducing a comparatively high titre in the corresponding group of rabbits. Similarly, comparatively poor responses in sheep were associated with poor responses in rabbits. The degree of variation in response within groups of animals was greater in sheep than in rabbits for all five antigenic components assayed. Sheep consistently developed higher titres than rabbits to Cl oedematins component but consistently lower titres to both Cl welchii beta and epsilon components irrespective of the type of vaccine used. The response of both species to Cl tetani antigen was similar in terms of serum antitoxic values. It was concluded that rabbits provide a suitable model for the assessment of potency of sheep clostridial vaccines.  相似文献   

15.
The possession of common antigens by three trematode parasites which commonly occur together in ruminants in the tropics, Fasciola gigantica, Dicrocoelium hospes and Schistosoma bovis was studied in relation to the reliability of serodiagnosis of infection with these helminths. The crude antigenic extracts of the three trematodes were subjected to Sephacryl S-300 column chromatography and F. gigantica was fractionated into six peaks, S. bovis into nine peaks and D. hospes into seven peaks. Common antigens were found in these three trematodes in both the crude whole worm antigenic extracts and in the semi-purified fractions obtained by Sephacryl S-300 column chromatography. The implications of this finding and the limitation it imposes on the usefulness of serodiagnostic tests in routine use as regards their specificity are discussed in relation to previous studies.  相似文献   

16.
A saline extract of a homogenate of Taenia saginata proglottides was partially purified by gel filtration chromatography on Sephadex G200 or Sepharose 4B and by ion exchange chromatography on DEAE cellulose. Gel filtration produced two distinct fractions with different antigenic properties. The first was of molecular weight of approximately 1,000,000 and contained a high level of activity in the haemagglutination inhibition test. The second fraction of molecular weigh of approximately 100,000 contained most of the immuno-precipitin activity. Other fractions had little or no antigenic activity. Eight fractions were obtained by DEAE cellulose chromatography, of which 4 had detectable antigenic activity. Subsequent rechromatography of fractions obtained by gel filtration on DEAE cellulose produced relatively pure fractions of high antigenic activity, from which small molecular weight contaminants had been removed.  相似文献   

17.
In the aim of improving serodiagnosis of canine leishmaniosis, we analysed the humoral immune response of dog against Leishmania infantum parasite. The antigenic reaction of L. infantum polypeptides with sera from 31 dogs with parasitologically confirmed leishmaniosis was studied by using the immunoblot technique. Electrophoretic profile of the parasite extract showed more than 50 polypeptides, with molecular weights ranging from 12 to 170 kDa. Among these polypeptides, 37 antigen components, ranging from 14 to 91 kDa, were recognised by antibodies of L. infantum infected dogs. Three polypeptides (14, 16 and 76 kDa) reacted with all of the 31 serum samples. The other most frequently recognised antigens were those of 29.5, 32, 46, 59 and 66 kDa with a sensitivity of 87.1%, 93.6%, 96.8%, 87.1% and 80.6%, respectively. The 14 and 16 kDa bands were the most intense and remained detectable until a serum dilution of 1:6400. No reaction of these two major antigens was observed with sera collected from 50 Leishmania-free dogs, living in the leishmaniosis-free region of Rabat in Morocco, whereas the crude antigen used in IFAT or ELISA lead to three false positive results. Four antigen components of 29, 41, 55, and 70 kDa were recognised by some sera samples from negative controls. These results demonstrated the potential interest of the fractions of 14 and 16 kDa in immunodiagnosis of canine leishmaniosis.  相似文献   

18.
利用浓度梯度SDS-聚丙烯酰胺凝胶电泳对猪囊尾蚴的囊液和虫体,猪带绦虫成虫的头颈节、成节、孕节,猪带绦虫的虫卵以及羊囊尾蚴的蛋白成分进行了对比分析。结果发现:囊液、虫体、头颈节、成虫、孕节、虫卵、羊囊尾蚴分别有35条(15.0-150.4kd)、44条(15.0-133.7kd)、22条(13.3-163.9kd)、19条(13.3-90.1kd)、19条(13.3-90.1kd)、30条(13.3-99.4)和38条(15.0-133.4kd)蛋白带;其中15.0kd和35.5kd两条共同抗原蛋白带,在猪带绦虫各阶段和羊囊尾蚴中含量最大的蛋白带同时也是阶段性的共同抗原蛋白带。这些结果为下一步的保护性抗原筛选奠定了基础。  相似文献   

19.
The kinetics of the antigen production of Chlamydia psittaci strains Izawa-1 and Pigeon-1041 (P-1041) was examined every 6 hr after infection up to 48 hr, by the indirect immunofluorescent antibody technique using monoclonal antibodies (MAbs). All three genus-specific antigenic determinants on lipopolysaccharide (LPS) appeared during the whole growth cycle. Antigenic determinants on proteins were, on the other hand, detected at various time periods from the early to the late stages of infection. However, a cross-reactive antigenic determinant on protein recognized by a MAb 3E9 was also detected during the whole growth cycle, similar to that on LPS. The time of appearance of common antigenic determinants on proteins of Izawa-1 and P-1041 was examined using cross-reactive MAbs, and it varied depending on heterologous and homologous MAbs. From the relationship between the detection of antigenic determinants and the morphological changes of chlamydial particles revealed by electron microscopy during the growth cycle, the antigenic determinants on proteins of Chlamydia psittaci were divided into two groups; one was specific to the elementary body and the other was coexisting in both the elementary body and the reticulate body.  相似文献   

20.
Hydatid cyst fluid from Echinococcus granulosus (HCF) and cyst fluid from Taenia hydatigena (TCF) cysts were compared in reciprocal immunoelectrophoresis (IEP) tests using homologous and heterologous antisera which were free of antibodies to host serum contaminants. The antigens for the E granulosus arc 5 were demonstrated in TCF. Antibody activity to these and other antigens common to HCF and TCF was removed from homologous antisera by absorptions with the heterologous antigenic preparation. Antigens not shared by the two metacestodes fluids were then demonstrated by IEP tests. These findings are discussed in terms of their significance to phylogenetic and immunodiagnostic studies of these parasites in their immediate hosts.  相似文献   

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