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1.
Localization of brucella antigens that elicit a humoral immune response in Brucella abortus-infected cells. 总被引:2,自引:1,他引:2
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Localization of brucella antigens, to which brucella-infected cattle make antibodies, and the surface characteristics of Brucella abortus smooth strain 19 and rough strain 45/20 were studied by the use of monospecific antisera in absorption tests, electron microscopy, and electrophoretic mobility of organisms in microelectrophoresis. Antigenic determinants of electrophoretically defined antigen A5 were present on the surface of B. abortus rough strain 45/20 organisms, and protein moieties were most probably exposed on the surface of this strain in contrast with smooth strain 19 organisms. Several antigens distinct from the smooth lipopolysaccharide complex, to which brucella-infected cattle make antibodies, were not detected on the surface of smooth organisms. Agglutinating antibodies present in anti-B. abortus strain 19 serum did not bind to all areas on the surface of the smooth cells, suggesting the presence of different antigenic moieties on their surface. It is also postulated that the surface of B. abortus rough strain 45/20 displays receptors able to strongly bind immunoglobulin molecules, as well as other serum components. 相似文献
2.
Antibody response to Brucella ovis outer membrane proteins in ovine brucellosis. 总被引:5,自引:4,他引:5
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Hot saline extracts of Brucella ovis were composed of vesicles with outer membrane proteins (OMPs), lipopolysaccharide, and phospholipid as constituents. Extraction with petroleum ether-chloroform-phenol yielded a protein fraction free of detectable lipopolysaccharide, in which group 3 OMPs (28,500 apparent molecular weight [28.5K], 27.0K, and 25.5K) represented 81% of the total. Group 1 OMPs and 67.0K, 22.5K to 21.5K, and 19.5K to 18.0K proteins were also detected. Adsorption of immune sera with whole bacteria suggested that group 3 OMPs and 67.0K, 22.5K to 21.5K, and 19.5K to 18.0K proteins had antigenic determinants exposed on the surfaces of both B. ovis and rough B. melitensis cells but not on smooth B. melitensis cells. Antibodies to group 3 OMPs and the 67.0K protein in the sera of 93 and 87%, respectively, of B. ovis-infected rams were found by immunoblotting. Antibodies to other proteins were present in 67% of these animals. Compared with B. ovis-infected rams which had not developed lesions, rams with epididymo-orchitis had antibodies to a larger variety of proteins. Although ewes infected with B. melitensis also showed antibodies to OMPs, the immunoblot reactions were less intense. 相似文献
3.
Enzyme-linked immunosorbent assay with major outer membrane proteins of Brucella melitensis to measure immune response to Brucella species.
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S B Hunter W F Bibb C N Shih A F Kaufmann J R Mitchell R M McKinney 《Journal of clinical microbiology》1986,24(4):566-572
We developed an enzyme-linked immunosorbent assay (ELISA) system to measure human immunoglobulin G (IgG) and IgM response to the major outer membrane proteins of Brucella melitensis. The ELISA was more sensitive in detecting antibody than a standard microagglutination (MA) test with B. abortus antigen. Of 101 sera from persons with suspected brucellosis, 79 (78.2%) gave ELISA IgM titers greater than or equal to the B. abortus MA titer without 2-mercaptoethanol (2ME), which measures both IgM and IgG. Of the 101 sera, 97% gave ELISA IgG titers greater than or equal to the MA with 2ME titer. A total of 58 sera, drawn from 11 human patients from 1 to 29 weeks after onset of brucellosis, gave higher geometric mean titers for the ELISA IgG test than for the MA with 2ME test. These 58 sera also gave ELISA IgM geometric mean titers that were greater than or within one doubling dilution of the geometric mean titers of MA without 2ME. In addition to detecting antibody response to B. abortus, B. melitensis, and B. suis, the ELISA was sensitive to antibody response to human and canine infections with B. canis. The B. canis antibody response is not detected by the MA test with B. abortus antigen. The ELISA, with a standard preparation of major outer membrane proteins of B. melitensis as antigen, appears to be useful in measuring antibody response in humans to infections by all species of Brucella known to infect humans. 相似文献
4.
Administration of monoclonal antibodies (mAb) to CD3 elicits an immune response to the mAb and an acute toxic syndrome that has been attributed to the release of cytokines from activated T cells. To clarify the cellular basis for these effects, we used anti-lymphocyte mAb to deplete selected T-cell subsets from BALB/c mice prior to administration of anti-CD3. In our first series of experiments, anti-CD4 repeatedly blocked the immune response to anti-CD3, but did not prevent severe toxicity. This observation suggested that other T-cell subsets might contribute to anti-CD3 induced toxicity. Therefore, we treated mice with mAb to CD8 as well as mAb to CD4 prior to administration of anti-CD3. Despite depletion of > 95% of CD8+ and CD4+ T cells, toxicity was not suppressed. This finding cast doubt on the belief that toxicity is due to activation of either CD4+ or CD8+ T cells by anti-CD3. Therefore, we assessed the role of thymocytes (which are not deleted by the mAb) and gamma delta + T cells. Thymectomy did not prevent toxicity in CD4/CD8-depleted mice, demonstrating that thymocytes are not responsible for toxicity. Anti-alpha beta TCR mAb produced a toxic reaction similar to anti-CD3 whereas anti-gamma delta TCR mAb did not, suggesting that gamma delta+ T cells are not the source of toxic cytokines. In addition, we proved that anti-CD3-induced toxicity was not due to direct effects on macrophages or to other nonspecific factors associated with the hamster mAb. These findings imply that a few residual mature T cells in mice treated with mAb to CD4 and CD8 are sufficient for the full expression of the anti-CD3-induced toxic syndrome. To confirm that both CD4+ and CD8+ T cells can mediate toxicity, we showed that:(i) SCID mice, which normally do not develop anti-CD3-induced toxicity, can be rendered susceptible by reconstitution with purified CD4+ T cells; and (ii) CD4-knockout mice that lack CD4+ T cells but have normal CD8+ T cells are susceptible to anti-CD3-induced toxicity. These findings establish that both CD4+ and CD8+ cells contribute to the toxic effects of anti-CD3, and that relatively few cells are required to mediate the full effect. 相似文献
5.
To test whether antibiotic therapy hampers the antibody response to Brucella antigens, 30 BALB/c mice were infected with Brucella melitensis H38 and randomized for treatment with doxycycline administered intraperitoneally for 42 days starting at 7 or 28 days postinfection (p.i.) (groups DOX7 and DOX28, respectively) or for no treatment (control group). Antibodies to smooth lipopolysaccharide (LPS) reached peak levels (mean optical density [OD] = 2.618) between days 56 and 70 p.i. in the control group, and similar peak levels (mean OD = 2.486) were observed in the DOX28 group, but significantly lower peak levels (mean OD = 0.821) were observed at 28 days p.i. in the DOX7 group. The antibody response against cytoplasmic proteins depleted of LPS (CPs) reached maximal levels (mean OD = 2.402) between days 56 and 70 p.i. in the control group, but no response was detected in the DOX7 group. Anti-CP antibodies were detected in only three animals from the DOX28 group, at levels significantly lower than those in the control group (mean maximal OD = 0.791). The pattern of antibody response to an 18-kDa cytoplasmic protein of Brucella spp. was similar to that against the CP antigen. This study shows that early antibiotic treatment affects the antibody response of mice to cytoplasmic proteins of Brucella and, to a lesser extent, to LPS. 相似文献
6.
T-cell subsets in autoimmunity. 总被引:6,自引:0,他引:6
The demonstration that functionally different T-cell subsets can be defined by the isoforms of the leukocyte-common antigen, CD45, that they express, has prompted studies on the roles of these subsets in autoimmunity. The results have led to the identification of a particular subset of CD4+ T cells that have the ability to inhibit autoimmune disease. Further, it has been shown that diabetes in the B-B rat can be transferred by in vitro activation of T cells by Staphylococcal enterotoxin suggesting that superantigens may play a role in the pathogenesis of this disease. However, in this system too, it appears that a subset of T cells can inhibit the induction of autoaggressive cells. In other experimental autoimmune diseases there is evidence that CD8+ T cells can be protective and that these cells may mediate this protection by the synthesis of transforming growth factor-beta. 相似文献
7.
T-cell lymphokine response to orally administered proteins during priming and unresponsiveness. 总被引:3,自引:0,他引:3
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Feeding antigens induces an immunological unresponsiveness termed oral tolerance but under some conditions, for example following the administration of cyclophosphamide (CY), immunity can be induced. These observations have usually been made by studying antibody production and delayed hypersensitivity with little attention given to other measurements of cellular activation. We have therefore examined the lymphokines produced by T cells obtained after the induction of oral tolerance or intragastric priming. Cells isolated from the spleen and Peyer's patches (PP) of tolerized mice could secrete high levels of interferon-gamma (IFN-gamma) and moderate levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) in response to antigen while interleukin-2 (IL-2), IL-3 and IL-4 could not be detected. Mesenteric lymph node (MLN) cells of tolerized mice did not respond to antigen unless spleen adherent cells were added to the cultures where IFN-gamma and GM-CSF were produced. Intragastric priming was achieved by feeding antigen to CY-treated mice. T cells from the spleen, MLN and PP of these mice could produce GM-CSF, IFN-gamma, some IL-3 but little or no IL-2 and IL-4. The ability of MLN cells to proliferate with antigen in vitro was low and corresponded to low IL-2 production. Thus T cells from fed mice secrete a defined pattern of lymphokines which differs in tolerizing and priming regimes. 相似文献
8.
F J Ramalho-Pinto O L Goldring S R Smithers H L Playfair 《Clinical and experimental immunology》1976,26(2):327-333
It is believed that a T-cell helper response against the schistosome surface is a necessary prerequisite for the development of protective immunity in schistosomiasis. Accordingly, the carrier effect has been used to assay eleven antigenic preparations of Schistosoma mansoni for their helper T-cell priming against surface components of the schistosomula. Three weeks after i.v. injection of the preparation, CBA mice were injected with schistosomula coated with trinitrophenol (TNP) and 4 days later, their spleens were assayed for plaque-forming cells (PFC) against TNP. Formalin-fixed schistosomula and crude adult worm tegumental membrane induced the highest response; only thirty schistosomula or 10mug of crude membrane protein were needed to generate a T-cell helper response equivalent to that induced by a living infection. All other antigenic preparations, including formalin-fixed cercariae, live miracidia, eggs and adult worm culture fluid, generated some response indicating the presence of shared carrier components. 相似文献
9.
Recombinant soluble tumor necrosis factor receptor proteins protect mice from lipopolysaccharide-induced lethality. 总被引:9,自引:0,他引:9
W Lesslauer H Tabuchi R Gentz M Brockhaus E J Schlaeger G Grau P F Piguet P Pointaire P Vassalli H Loetscher 《European journal of immunology》1991,21(11):2883-2886
The in vivo efficacy of human recombinant soluble tumor necrosis factor (TNF) receptor protein to prevent and to treat lipopolysaccharide (LPS)-induced lethal toxicity in D-galactosamine-treated mice was investigated. Chimeric proteins of the receptor extracellular domains fused to the hinge region of human IgG3 were expressed in myeloma cells (rsTNFR-h gamma 3). The fusion proteins had a disulfide-bonded dimeric structure. Upon intravenous injection, their serum concentration decreased relatively slowly after an initial phase of rapid elimination. D-galactosamine-sensitized mice were fully protected from the toxic effects of LPS, if the animal were pretreated with rsTNFR-h gamma 3 at 20 micrograms/animal. Partial protection was seen at significantly lower doses and when rsTNFR-h gamma 3 was given up to 3 h after LPS. 相似文献
10.
Thymopentin reduces the susceptibility of aged mice to cutaneous leishmaniasis by modulating CD4 T-cell subsets.
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E Cillari S Milano M Dieli F Arcoleo R Perego F Leoni G Gromo A Severn F Y Liew 《Immunology》1992,76(3):362-366
BALB/c mice are highly susceptible to Leishmania major infection. The susceptibility increases progressively with the age of the mice. Aged mice produce progressively lower levels of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) but higher levels of IL-4 compared to younger mice. Thymopentin, a pentapeptide with thymopoietin activity, dramatically increases the resistance to Leishmania major infection in aged mice. The thymopentin-treated mice produce enhanced levels of IL-2 and IFN-gamma, but significantly reduced amounts of IL-4. Thus, it appears that the age-related susceptibility to cutaneous leishmaniasis is correlated with the enhancement of Th2 and the reduction of Th1 cell activities. Furthermore, thymic hormone may play an important role in the induction and function of these two subsets of CD4 T cells. 相似文献
11.
Declining immunity in aged individuals has been repeatedly verified. However, many of the studies have focused on functions not directly relevant to the antigens normally encountered in vivo. For example, although infectious agents represent a significant cause of morbidity and mortality in the elderly, age-related immune responses in such diseases have received very little experimental attention. The present study was, therefore, initiated to examine T cell-mediated immunity to influenza virus in aged mice. We observed that the primary cytotoxic T lymphocyte (CTL) response to influenza, which peaks at day 5 in young adult mice, is almost completely absent at that timepoint in aged mice. Our experiments indicate that the kinetics of the response are shifted, so that the maximum activity appears at days 7-9. Furthermore, a comparison of the peak responses of both age groups reveals a significantly lower magnitude in the aged mice. The diminished response was also observed for most of the old mice when antigenic stimulation occurred in vitro. Although the overall response was diminished, the characteristic T cell cross-reactivity for all A strain viruses was similar to that observed in young mice. The possible causes and consequences of the age-related decline in influenza-specific CTL activity are discussed. 相似文献
12.
Ultrastructural morphometric analysis of Brucella abortus-infected trophoblasts in experimental placentitis. Bacterial replication occurs in rough endoplasmic reticulum. 总被引:6,自引:0,他引:6
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Trophoblasts in normal and Brucella abortus-infected caprine placentas were examined by ultrastructural morphometric analysis to establish structural relationships of B abortus with cytoplasmic organelles; brucellae were identified with colloidal gold-labeled anti-B abortus bovine IgG. Cytotrophoblasts had large numbers of B abortus in cisternae of rough endoplasmic reticulum; binucleate trophoblasts did not contain bacteria. In infected trophoblasts there was a significant hypertrophy of B abortus-filled rough endoplasmic reticulum (RER) and a corresponding reduction in normal-appearing RER. Volume and surface density of RER in trophoblasts were: normal placentas (control), 2.8% and 0.30 sq mu; infected placentas, 27.9% (27.4% of which contained B abortus) and 0.56 sq mu (cells containing B abortus) and 3.3% and 0.34 sq mu (cells not containing B abortus). These data suggest that B abortus replicates within the RER of trophoblasts, possibly for synthesis and glycosylation of bacterial membrane proteins. 相似文献
13.
Regulation of macrophage phagocytosis of syngeneic erythrocytes by T-cell subsets from NZB mice: differential effects of T cells from young and old mice.
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The regulation of syngeneic erythrophagocytosis (EP) by macrophages (M phi) harvested from young and old NZB mice was examined by spectrophotometric assay and morphological observation. Peritoneal exudate M phi from young NZB mice weakly ingested syngeneic red blood cells (RBC). T cells derived from old NZB mice accelerated ingestion of RBC by young M phi. On the contrary, T cells from young NZB mice suppressed EP by young T cells appeared clearly when they were added to M phi derived from old mice, which ingested syngeneic RBC actively without help by old NZB T cells. Namely, such an active EP by old M phi was completely suppressed when they were incubated with young T cells. Simultaneous addition of both young and old T cells to either young or old NZB M phi with RBC suppressed the EP. Pretreatment of young T cells with anti-Lyt 1.2 antibody and complement (C) made the suppressive activity prominent, and preincubation with anti-Lyt 2.2 and C eliminated the suppressive activity, but gave rise to the enhancing activity. Young T-cell homogenates added to younger or old M phi together with RBC did not reveal suppressive activity for EP, and on the contrary facilitating activity appeared predominantly. Young and old T-cell homogenates added together to young M phi did not suppress EP. The largest of T-cell-factor accelerating EP was M phi, but not RBC. M phi with active EP belong to Ia-bearing subpopulations. 相似文献
14.
For analysis of the role of immune T cells in protective immunity against murine malaria, Plasmodium yoelii-immune Lyt T-cell subsets were functionally characterized in vitro and in vivo. Selected Lyt2- and Lyt2+ T cells from P. yoelii-immune C57BL/10 mice differed in their capability to proliferate in response to P. yoelii antigen in vitro. Only the Lyt2- T-cell population produced T-cell growth factor upon restimulation, and none of the selected T-cell subsets produced detectable amounts of macrophage activating factor. Lyt2- but not Lyt2+ lymphocytes were capable of transferring protection to normal C57BL/10 mice. When transferred into T-cell-deficient C57BL/6-nu/nu mice, adoptive resistance to P. yoelii by Lyt2- lymphocytes was only demonstrable after prior reconstitution of recipients with normal T cells. These results suggest an interaction between P. yoelii-immune Lyt2- T cells and normal T lymphocytes via T-cell growth factor in the development of protective immunity to malaria. 相似文献
15.
Variation of Brucella abortus 2308 infection in BALB/c mice induced by prior vaccination with salt-extractable periplasmic proteins from Brucella abortus 19.
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The study compared the immune and protective responses induced in BALB/c mice vaccinated with six salt-extractable periplasmic protein fractions (Brucella cell surface proteins [BCSP]) of Brucella abortus 19 and later challenge exposed with B. abortus 2308. BCSP70 was precipitated with ammonium sulfate at 70% saturation, and BCSP100 was precipitated with ammonium sulfate at 100% saturation by use of supernatant fluid of BCSP70 that had been precipitated with 70% ammonium sulfate. Four subfractions were separated from BCSP100 by anion-exchange high-performance liquid chromatography (HPLC). Monophosphoryl lipid A (MPL) from Salmonella typhimurium Re mutant strain was used as a potential immune response modifier in some vaccines. Reduced or increased numbers of CFU and increased spleen size in the principal groups of mice relative to that of the nonvaccinated control group were considered protectiveness or virulence (survival) criteria. Results indicated that vaccines prepared from BCSP70 and BCSP100 were moderately protective and immunogenic. The subfractions designated BCSP100-A through BCSP100-D purified by anion-exchange HPLC were not protective when MPL was not used as an immune response modifier. However, two subfractions were associated with significant (P < 0.05) increases in CFU per spleen and splenomegaly in vaccinated mice compared with those in nonvaccinated challenge-exposed mice. MPL enhanced protection or was neutral when used with BCSP70, BCSP100, BCSP100-C, and BCSP100-D. Serologic results of an enzyme-linked immunosorbent assay indicated that MPL modulated the immunoglobulin G responses induced by BCSP70, BCSP100, and subfraction BCSP100-B vaccines only. The overall results suggest that certain proteinaceous periplasmic fractions might serve as virulence or survival factors in B. abortus infections. 相似文献
16.
Comparison of spleen cell proliferation in response to Brucella abortus 2308 lipopolysaccharide or proteins in mice vaccinated with strain 19 or RB51.
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Mice vaccinated with Brucella abortus 19 (S19) or RB51 (SRB51) had spleen cells which proliferated in response to proteins of 32, 27, 18, and < 18 kDa but not in response to proteins of 106, 80, and 49 kDa from B. abortus 2308 (S2308) following vaccination and challenge infection with S2308. Spleen cells from mice vaccinated with S19 but not with SRB51 had increased proliferation in response to S2308 lipopolysaccharide (LPS) following challenge infection with S2308. We previously reported that mice vaccinated with S19 or SRB51, which were analyzed in the current study, have increased resistance to infection with S2308 and that only mice vaccinated with S19 produce antibody to S2308 LPS (M. Stevens, S. Olsen, G. Pugh, Jr., and D. Brees, Infect. Immun. 63:264-270, 1995). The results from our current and previous studies support the contention that vaccination of mice with S19 or SRB51 induces protection from infection with S2308 by cell-mediated immune responses to the same immunodominant (32, 27, 18, and < 18 kDa) protein antigens of S2308. In addition, the absence of S2308 LPS-responsive spleen cells and antibody to S2308 LPS in mice vaccinated with SRB51 suggests that immune responses to LPS have no role in SRB51-induced protective immunity. 相似文献
17.
目的 研究小鼠注射HIV gag DNA疫苗后的抗原特异性细胞免疫应答.方法 C57BL/6小鼠以初免/加强的策略经肌肉注射HIV gag DNA疫苗,一周后获取其脾与肺的单个细胞,体外经Gag抗原多肽刺激后,采用ELISA法检测细胞培养上清中IFN-γ的水平,ELISPOT法检测IFN-γ分泌细胞的频率,流式细胞仪分析特异性T细胞的亚群.结果 经Gag多肽刺激后,加强免疫组IFN-γ产生的总体水平和分泌细胞频率均高于对照组及初次免疫组.HIV gag DNA疫苗可同时诱导产生Gag-特异性CD4 和CD8 T细胞.结论 HIV gagDNA疫苗免疫小鼠后可诱导抗原特异性效应性T细胞应答. 相似文献
18.
Outer membrane proteins from rough strains of four Brucella species. 总被引:9,自引:18,他引:9
Outer membrane proteins from 15 rough strains of Brucella abortus, B. ovis, B. canis, and B. melitensis were extracted with a dipolar detergent, and outer membrane proteins from selected strains were purified by anion exchange chromatography and gel filtration (Verstreate et al., Infect. Immun. 35:979-989, 1982). Outer membrane proteins produced two types of profiles on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. One type, demonstrated by B. abortus, B. ovis, and B. canis strains, contained the three predominant protein groups present in smooth B. abortus strains (Verstreate et al., Infect. Immun. 35:979-989, 1982): groups 1, 2 (porin [Douglas et al., Infect. Immun. 44:16-21]), and 3. B. melitensis strains demonstrated the second profile type, in which there was an additional band between groups 1 and 2. The relative proportion of porin was considerably lower in B. ovis, B. canis, and B. melitensis than in B. abortus. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles could be used to distinguish B. abortus and B. melitensis from each other and from B. canis and B. ovis. The amino acid compositions of groups 2 and 3 from rough strains of B. abortus, B. canis, and B. melitensis were similar to those of corresponding proteins from smooth B. abortus strains. Zwittergent-soluble fractions from most rough strains contained antigen [b], which cross-reacted with group 2 from smooth B. abortus strains, and antigens [c] and [d], which cross-reacted with group 3 from smooth B. abortus strains. Antigen [a], shared by groups 2 and 3 (D. R. Verstreate and A. J. Winter, Infect. Immun. 46:182-187, 1984), was detected in most rough strains. None of these antigens were related to either rough or smooth lipopolysaccharide. 相似文献
19.
Bone is a dynamic tissue with a well-balanced homeostasis preserved by both formation and resorption of bone. Normal turnover of bone, however, can be upset by either increased osteoclast activity or decreased osteoblast function; either mechanism alone or both may result in a net loss of bone. Both osteoclasts and osteoblasts could be stimulated by mechanical stimulation in vitro, and it is assumed that this process may occur in vivo as well. In this experiment, we investigated this hypothesis by examining the effects of ultrasound stimulation on osteoblast growth and cytokine release. With this model, we explored the mechanism of low-intensity pulsed ultrasound on osteoblasts growth and upregulation of osteoclasts formation and function by cytokine release. The results showed that specific pulsed ultrasound exposure could enhance osteoblasts population together with increase in TGFbeta1 secretion and decrease in concentration of IL-6 and TNFalpha in the culture medium. Although, animal studies and clinical trial are needed to understand the real process in the whole body, ultrasound stimulation might be a good method for prevention of bone loss due to osteoporosis. 相似文献
20.
Antibody response to antigens distinct from smooth lipopolysaccharide complex in Brucella infection. 总被引:5,自引:8,他引:5
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The smooth lipopolysaccharide complex of the outer surface of smooth Brucella abortus cells is believed to be the antigenic component involved in serological tests routinely used for the diagnosis of brucellosis. Sera from cattle vaccinated or infected with B. abortus generally contain antibody directed toward the smooth lipopolysaccharide complex. The brucella organism contains a large number of other antigenically distinct components. The biological significance of some of these antigens has been demonstrated by showing that sera from infected cattle have precipitins to these components. These sera revealed up to seven distinct lines in immunoelectrophoresis with a protein-rich antigen mixture prepared from rough strain B. abortus 45/20, whereas sera from strain 19-vaccinated cattle did not reveal these lines at 4 or more months after vaccination. Monospecific antisera were prepared against six antigens in this mixture, and the purification of two of them by antibody affinity chromatography is described. 相似文献