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1.
Hereford cattle protected against Boophilus microplus with antigens purified by immunoaffinity chromatography from larval and adult ticks. 总被引:4,自引:0,他引:4
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The subpopulations of lymphocytes in the pregnant and non-pregnant mammary glands of the sheep were delineated by a panel of monoclonal antibodies. The most striking feature observed was that in the mammary gland of both pregnant and non-pregnant sheep the great majority of the lymphocytes in the ductal and alveolar epithelium were agranulated CD8+ CD5- cells. A small subpopulation of granulated lymphocytes in the epithelium expressed the CD45R antigen but not the major histocompatibility complex (MHC) class II molecules. Other subpopulations, especially B lymphocytes, were present in much lower concentrations and were located mainly in the periductal and intralobular connective tissues. Patches of lymphocytes clustering around venules were observed and the majority of them were shown to be CD5+ CD4+, while some were CD5+ CD8+ but none were CD45R+ (B cell). It is suggested that selective traffic of T cells occurs at these sites. 相似文献
2.
Expression of equi merozoite antigen 2 during development of Babesia equi in the midgut and salivary gland of the vector tick Boophilus microplus
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Ueti MW Palmer GH Kappmeyer LS Scoles GA Knowles DP 《Journal of clinical microbiology》2003,41(12):5803-5809
Equi merozoite antigens 1 and 2 (EMA-1 and EMA-2) are Babesia equi proteins expressed on the parasite surface during infection in horses and are orthologues of proteins in Theileria spp., which are also tick-transmitted protozoal pathogens. We determined in this study whether EMA-1 and EMA-2 were expressed within the vector tick Boophilus microplus. B. equi transitions through multiple, morphologically distinct stages, including sexual stages, and these transitions culminate in the formation of infectious sporozoites in the tick salivary gland. EMA-2-positive B. equi stages in the midgut lumen and midgut epithelial cells of Boophilus microplus nymphs were identified by reactivity with monoclonal antibody 36/253.21. This monoclonal antibody also recognized B. equi in salivary glands of adult Boophilus microplus. In addition, quantification of B. equi in the mammalian host and vector tick indicated that the duration of tick feeding and parasitemia levels affected the percentage of nymphs that contained morphologically distinct B. equi organisms in the midgut. In contrast, there was no conclusive evidence that B. equi EMA-1 was expressed in either the Boophilus microplus midgut or salivary gland when monoclonal antibody 36/18.57 was used. The expression of B. equi EMA-2 in Boophilus microplus provides a marker for detecting the various development stages and facilitates the identification of novel stage-specific Babesia proteins for testing transmission-blocking immunity. 相似文献
3.
Protective efficacy of antigens solubilized from gut membranes of the cattle tick, Boophilus microplus
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Triton X-100, SDS, n-octyl glucoside, and conditions of alkaline pH and high and low ionic strength, were used to solubilize membranes prepared from the mid-gut of the cattle tick Boophilus microplus. Hereford cattle (Bos taurus) were immunized with soluble membrane antigens in saponin, and then challenged twice with 20,000 tick larvae, 7 days apart. All soluble extracts, except that extracted with high ionic strength buffer, provided significant protection (56-78%) (P < 0.05) to immunized cattle compared with control cattle. Precipitation of the soluble n-octyl glucoside extract did not alter the immunogenicity of this vaccine. Where poor responses to membrane vaccines were obtained (< 60% protection), booster injections of antigen effected increased protection. The experimental cattle were typed for bovine major histocompatibility system class 1 antigens. The presence of three antigens (MB5, MB14 and MB20) had a significant effect on protective immunity induced by the membrane vaccines (P < 0.05). Protein profiles of Triton X-100, n-octyl glucoside and low ionic strength buffer extracts were compared by high-performance size-exclusion chromatography. Tick membranes extracted by either non-ionic detergents or low ionic strength solutions provided the best source of protective antigens. 相似文献
4.
To develop immunoprophylactic measures against multi-tick infestation, two glycoproteins of 34 and 29 kDa were isolated from the larvae of Hyalomma anatolicum anatolicum and Boophilus microplus, respectively, and assessed for their efficacy against experimental challenge infestations. The synergistic effect of the antigens in the presence of incomplete Freunds adjuvant was found to confer protection (DT%) in animals against 56.48% of larvae and 52% of adults of H. a. anatolicum, while the effect was 40% against adults of B. microplus. The efficacy (E%) of the antigens in combination against larvae and adults of H. a. anatolicum was calculated as 70% and 64.3%, respectively, and 63% against adults of B. microplus. A direct correlation between anti-glycoprotein antibody response and protection against infestation was observed. Western blot analysis detected specific antigen in the sera of animals of group A. The antigens in combination with incomplete Freunds adjuvant could protect animals from H. a. anatolicum and B. microplus infestations for at least 30 weeks. The possibility of employing the vaccination strategy in Indian conditions is discussed. 相似文献
5.
DNA vaccines provide an attractive technology against cancer because of their safety record in humans and ease of construction, testing and manufacture. In this study, several DNA fragments encoding multiple cytotoxic T lymphocyte (CTL) and T helper cell epitopes were selected from human prostate-specific membrane antigen (hPSM), mouse prostatic acid phosphatase (mPAP), and human prostate-specific antigen (hPSA). These DNA fragments were ligated together to form a novel fusion gene, termed the 3P gene. The secondary lymphoid tissue chemokine (SLC), 3P and human immunoglobulin G Fc genes were inserted into pcDNA3.1 to construct a DNA vaccine, designated pSLC-3P-Fc. After vaccination, the DNA is taken up by cells that produce and secrete the SLC-3P-Fc fusion proteins, termed chemotactic antigen (chemo-antigen). The secreted chemo-antigens, in addition to promoting the co-localization of naive, non-polarized memory T cells and dendritic cells, are efficiently captured and processed by dendritic cells via receptor-mediated endocytosis and then cross-presented to both major histocompatibility complex class I and class II in a cognate manner. The results of this study demonstrate that vaccination with pSLC-3P-Fc by gene gun inoculation induced a strong antitumour response in a mouse tumour model, which significantly inhibited tumour growth and prolonged the survival time of the tumour-bearing mice. In vitro, the secreted SLC-3P-Fc fusion protein can attract lymphocytes from human peripheral blood mononuclear cells (PBMC); when human lymphocytes were stimulated by pSLC-3P-Fc-transfected autologous PBMC, CTLs were induced which could specifically kill hPSM-, hPAP-, or hPSA-expressing tumour cells. These observations provide a new vaccine strategy for cancer therapy through promoting the co-localization of lymphocytes and the concomitant enhancement of antigen-specific CD4+ helper and CD8+ cytotoxic T-cell responses against tumour. 相似文献
6.
Summary Cattle with differing degrees of resistance toBoophilus microplus have responses to tick allergen which correlate with their resistance level The total amount of histamine in the skin also correlates with both resistance and the immediate hypersensitivity reactions, but the sensitivity to injected histamine does not. Treatment with the antihistamine drug mepyramine maleate suppresses the cutaneous hypersensitivity reactions. The results suggest that the main pharmacologically active agent in these reactions is histamine, and that the total amount of it available locally in the skin may have a role in the resistance to this parasite. 相似文献
7.
Consuelo Almazán Rodolfo Lagunes Margarita Villar Mario Canales Rodrigo Rosario-Cruz Frans Jongejan José de la Fuente 《Parasitology research》2010,106(2):471-479
The cattle ticks, Rhipicephalus (Boophilus) spp., affect cattle production in tropical and subtropical regions of the world. Tick vaccines constitute a cost-effective
and environmentally friendly alternative to tick control. The recombinant Rhipicephalus microplus Bm86 antigen has been shown to protect cattle against tick infestations. However, variable efficacy of Bm86-based vaccines
against geographic tick strains has encouraged the research for additional tick-protective antigens. Herein, we describe the
analysis of R. microplus glutathione-S transferase, ubiquitin (UBQ), selenoprotein W, elongation factor-1 alpha, and subolesin (SUB) complementary
DNAs (cDNAs) by RNA interference (RNAi) in R. microplus and Rhipicephalus annulatus. Candidate protective antigens were selected for vaccination experiments based on the effect of gene knockdown on tick mortality,
feeding, and fertility. Two cDNA clones encoding for UBQ and SUB were used for cattle vaccination and infestation with R. microplus and R. annulatus. Control groups were immunized with recombinant Bm86 or adjuvant/saline. The highest vaccine efficacy for the control of
tick infestations was obtained for Bm86. Although with low immunogenic response, the results with the SUB vaccine encourage
further investigations on the use of recombinant subolesin alone or in combination with other antigens for the control of
cattle tick infestations. The UBQ peptide showed low immunogenicity, and the results of the vaccination trial were inconclusive
to assess the protective efficacy of this antigen. These experiments showed that RNAi could be used for the selection of candidate
tick-protective antigens. However, vaccination trials are necessary to evaluate the effect of recombinant antigens in the
control of tick infestations, a process that requires efficient recombinant protein production and formulation systems. 相似文献
8.
Live attenuated Francisella tularensis vaccine induced long-lasting humoral and cell-mediated immune responses in all 13 subjects studied. Lymphocyte blast transformation reactivity to F. tularensis appeared 2 weeks after vaccination in most subjects and remained unchanged for up to 1.5 years. Similarly, in most recipients, antibodies against F. tularensis were detectable by both the enzyme-linked immunosorbent assay (ELISA) and the agglutination method from 2 weeks after vaccination, although diagnostically significant agglutination titers (greater than or equal to 80) were not detectable until 4 weeks after vaccination. Maximal agglutination titers of 80 to 2,560 appeared at 4 to 8 weeks, and in spite of decreasing tendency, titers as high as 320 were still present 1.5 years after vaccination. ELISA showed the simultaneous, but not parallel, appearance of different immunoglobulin classes, immunoglobulin M (IgM) reaching individual maximal values 1.8 months after vaccination on average, at the same time as agglutinating antibodies, 1 week earlier than IgA, and about 1 month earlier than IgG. All of these immunoglobulin classes persisted in significant amounts up to 1.5 years, with IgG generally dominant. Long-lasting IgA and IgM responses after vaccination, as also after infection, suggested that the serodiagnosis of tularemia generally requires two consecutive serum samples with a significant increase in the titer. 相似文献
9.
A Czarny D Witkowska M Mulczyk 《Archivum immunologiae et therapiae experimentalis》1992,40(2):129-134
Shigella flexneri outer membrane proteins (OMP) which had been earlier found to exert immunomodulatory effect on cell mediated immune response were also found to act as immunomodulator of the humoral immune response. Effects of OMP were investigated in the experiments in vitro and in vivo, where the level of humoral immune response, measured as the number of plaque-forming cells (PFC) to SRBC in the spleen was evaluated. We demonstrate that small doses of OMP (1-5 micrograms) stimulate, whereas higher doses (10-50 micrograms) suppress the humoral immunity. 相似文献
10.
Duarte MO Ferrarini SR Pazinatto M de Oliveira ER Rolim V Eifler-Lima VL Ribeiro VL von Poser G 《Parasitology research》2008,103(3):723-726
This study reports the effect of six hyacinthacine analogues derived from pyrrolizidine alkaloids on egg hatchability and mortality rates of newly hatched larvae of the cattle tick Rhipicephalus (Boophilus) microplus. All the compounds were toxic to the larvae of the ticks and inhibited the eggs hatchability in the higher concentration tested (5 microg/ml). Even in the lowest concentration (0.625 microg/ml), some effect in the eggs hatchability was observed. 相似文献
11.
Histopathological study of protective immunity against murine salmonellosis induced by killed vaccine. 总被引:4,自引:2,他引:2
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Swiss-Webster mice were vaccinated with heat-killed salmonellae and then were infected with virulent Salmonella typhimurium. Only 1 of the 18 vaccinated mice died from a challenge of 10(4) X the 50% lethal dose, and about 70% of them survived a challenge of 10(5) X the 50% lethal dose. Histopathological examinations of the lesions developed in these vaccinated mice showed that they followed the characteristic features of a primary lesion in murine salmonellosis. There was an early necrosis with infiltration of polymorphonuclear leukocytes and abscess formation within the first 6 to 7 days after infection. However, these abscesses remained small and discrete. By days 7 to 10, the lesions began to transform into granulomas, first with the appearance of peripheral mononuclear cells and then by the replacement of polymorphs. By the third week of the infection, minute and discrete granulomas were seen scattered in the spleen, liver, and lymph nodes. Beyond this stage, healing and tissue regeneration followed. Thus, the characteristics of infectious lesions developed in mice vaccinated with heat-killed salmonellae are distinctly different from those developed in mice protected by the avirulent vaccine. 相似文献
12.
The effect of cytochalasin-B concentration on the frequency of micronuclei induced by four standard mutagens. Results from two laboratories 总被引:2,自引:6,他引:2
Surralles J.; Antoccia A.; Creus A.; Degrassi F.; Peris F.; Tanzarella C.; Xamena N.; Marcos R. 《Mutagenesis》1994,9(4):347-353
In a previous collaborative work, we have recently shown thatthe cytochalasin-B (Cyt-B) concentration used in the human lymphocytescytokinesis-block micronucleus (CBMN) assay is an importantvariable in the baseline micronuclei (MN) frequency as wellas in the percentage of binucleated cells obtained. Now we haveinvestigated how Cyt-B concentration modulates the MN frequencyinduced in whole blood human lymphocyte cultures by two clastogens(ethyl methanesulphonate and mitomycin-C) and two aneugens (colchicineand vincristine sulphate). The experimental design includessix donors, two concentrations of Cyt-B (3 and 6 µg/ml),two concentrations of the four chemicals tested and the exchangeof slides between laboratories. The statistical analysls ofthe results shows: (i) non-significant differences in the MNfrequencies and in the toxicity results between scorers fromeach laboratory, except for 0.06 µM colchicine at 3 µg/mlCyt-B; (ii) an induction of MN by all genotoxic agents tested,the frequencies being lower with 6 than with 3 µg/ml Cyt-B,in control and aneugen-treated cultures; and (iii) significantdifferences between Cyt-B concentrations in several treatments,obtaining lower MN frequencies and higher values for nucleardivision index and % binucleated cells when 6 µg/ml Cyt-Bwas used. Bearing in mind these results as well as the toxicitydata showing that 6 µg/ml Cyt-B is much more effectivein blocking cytokinesis, we can conclude that the use of 3µg/mlCyt-B may overestimate the induced frequency of MN.
4To whom correspondence should be addressed 相似文献
13.
14.
Autoimmunity induced by injection of virus-modified cell membrane antigens in syngeneic mice.
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C3H/Bi mice developed autoantibodies after repeated inoculations of isolated membranes from primary tissue cultures of a syngeneic ascites lymphoma in which Newcastle disease virus had grown. This was in addition to the tumor transplantation resistance and cytotoxic antibodies previously demonstrated. The complement-fixing antibodies were completely removed from sera by adsorption with ascites tumor cells but only partially by normal mouse liver powder or C3H/Bi erythrocytes. With continued immunization, antibodies to deoxynucleoprotein and heterophile reagins also appeared. After several months, mice showing these serological reactions died with a wasting disease characterized by loss of lymphoid tissue and scarred, atrophied kidneys. No significant antibody response or autoimmune disease occurred in mice receiving membranes from uninfected syngeneic ascites lymphoma. 相似文献
15.
目的 研究结核融合蛋白Ag85B-Mpt64190-198-Mth8.4(AMM)和佐剂二甲基三十六烷基铵(DDA)、卡介苗多糖核酸(BCG-PSN)构建的亚单位疫苗强化BCG初始免疫的免疫效应.方法 将融合蛋白AMM、佐剂DDA和BCG-PSN混合构建AMM亚单位疫苗.实验1组BCG初免后第10周用AMM亚单位疫苗加强免疫小鼠一次;实验2组BCG初免后分别于第8周、第10周用AMM亚单位疫苗加强免疫小鼠一次.同时设立生理盐水及仅BCG免疫两个对照组.BCG初免后第14周、第22周,应用ELISPOT、ELISA检测免疫小鼠的细胞及体液免疫反应.同时在第22周用BCG活菌攻击被免疫小鼠,间隔4周后用流式细胞术和ELISA技术检测T细胞分型及体液免疫反应.结果 (1)IFN-γ水平:BCG初免后14周,特异性抗原Ag85B刺激后实验2组分泌IFN-γ的细胞数(135±14)明显高于仅BCG免疫组(194±16),t=10.98,P<0.01;BCG初免后22周,实验2组(208±11)同样高于仅BCG免疫组(57±18),t=6.43,P<0.01.(2)体液免疫应答水平:实验2组的IgGI抗体滴度明显高于实验1组,而作为反映Th1型免疫反应指标的IgG2a/IgG1比值,强化免疫两次组低于强化一次组.(3)BCG模拟攻击被免疫小鼠后,CD4+CD25+调节性T细胞含量:实验1、2组均高于仅BCG免疫组(t1=3.08,t<2>=3.16,P<0.05).结论 BEG免疫-AMM亚单位疫苗加强免疫两次能够引起较强的细胞及体液免疫反应,同时激活调节性免疫反应. 相似文献
16.
Protective immunity induced by outer membrane proteins of Salmonella typhimurium in mice. 总被引:14,自引:10,他引:4
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Outer membrane proteins (OMP) extracted from both smooth (C5) and rough (Rb2) strains of Salmonella typhimurium were able to induce protective immunity to salmonellosis. The OMP-induced protection lasted for at least 6 months. The antibody level was estimated by passive hemagglutination. In the C5 OMP-immunized mice, antibodies to both proteins and lipopolysaccharide were detected. On the other hand, in the Rb2 OMP-immunized mice, antiprotein but not antilipopolysaccharide antibodies were detected. Delayed-type hypersensitivity appeared as early as the second week after immunization with OMP and persisted through the fourth week. 相似文献
17.
Analysis of immunity induced by the affinity-purified 21-kilodalton zygote-ookinete surface antigen of Plasmodium berghei. 总被引:1,自引:0,他引:1
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By using affinity-purified ookinete surface antigen from the rodent malaria parasite Plasmodium berghei, a transmission-blocking immunity was induced in mice. Groups of mice were immunized via different routes, with total quantities of antigen ranging from 0.5 to 40 micrograms (with or without Freund adjuvant). Vaccination by the intramuscular route with 20 micrograms of antigen in the absence of adjuvant and boosted once with the same amount of protein induced a total transmission blockade. Immunoblot analysis confirmed that immune sera invariably recognized Pbs21 antigen. The isotype and titer of the anti-Pbs21 immunoglobulin G (IgG) response was determined by enzyme-linked immunosorbent assay. The antibody isotype was predominantly IgG1. The concentration of specific anti-Pbs21 IgG reached a peak of 182.45 +/- 92.13 micrograms/ml by week 7 postimmunization and fell progressively to 38 micrograms/ml at week 34 (at which time the transmission was still inhibited by 98%). 相似文献
18.
19.
Fifty isolates of Beauveria bassiana (Balsamo) Vuillemin, 1912 (Ascomycota: Clavicipitaceae) were analyzed by morphology, for their pathogenic potential to Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) larvae, and by Random Amplified Polymorphic DNA-Polymerase Chain Reaction technique.
Morphological analysis demonstrated that isolates present characteristics compatible to those described for B. bassiana in the literature. Virulence test demonstrated that all isolates present lethal effect on larvae and that the lethal concentration
varies among isolates. The most virulent isolate was the only one obtained from human infection, which was also the only isolate
presenting synnemata. The study on genetic variability among the isolates allowed the identification of 23 electrophoretic
profiles. The established groupings suggest that most of the isolates obtained from B. microplus of the same locality present low genetic variation. In this way, the data in the present study will contribute to a meticulous
characterization of these B. bassiana isolates. 相似文献
20.
G G Klaus 《European journal of immunology》1976,6(6):389-393
Injection of mice already making antibodies to 2,4-dinitrophenylated (DNP) Ficoll with tolerizing doses of DNP-pneumococcal polysaccharide (DNP-lys-S3) markedly inhibits the secretion of anti-DNP antibodies by IgM antibody-forming cells. The present study shows that the degree of inhibition depends not only on the dose of DNP-lys-S3 but also on the duration of exposure to antigen. DNP-lys-S3 was detectable on the surface of antibody-forming cells at a time when their rate of secretion was unimpaired, thus suggesting that the inhibition involves intracellular events subsequent to the binding of antigen to the cell membrane. The inhibition was reversible if antibody-forming cells were exposed to antigen for 24 h, and then cultured for 18 h in its absence, but became irreversible if the treatment period was extended to 48 h. The relevance of this model of inhibition of lymphocyte function by antigen to possible mechanisms of B cell tolerance is discussed. 相似文献