The Cellular Immunoprotection of BALB/C mice vaccinated with Salt-Extractable Brucella abortus S19 antigens and Immunoadjuvant βeta-glucan challenged with Brucella abortus Virulent Strain (original) (raw)
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Infection and immunity, 1994
A study was conducted to determine whether the covalent chemical modification of Brucella abortus 19 salt-extractable proteins (BCSP) and BCSP derivatives would modulate the immune responses in BALB/c mice. Salt-extractable proteins BCSP 0-70 and BCSP 70-100 were modified with acetoacetic anhydride, and recombinant proteins rBCSP20 (20 kDa), rBCSP31 (31 kDa), and rBCSP45 (45 kDa) were modified with succinic and dodecanoyl anhydrides. Four weeks after mice were vaccinated with the different preparations, principal and control mice were challenge exposed with a virulent culture of B. abortus 2308, and mice were necropsied 2 weeks later. Serum samples were obtained immediately before mice were challenge exposed and at necropsy. Sera were tested for specific immunoglobulin M (IgM) and G (IgG) antibodies by using an enzyme-linked immunosorbent assay. Acylation decreased the immune responses (increased IgG antibodies and reduced spleen CFU and splenomegaly) induced by both BCSP 0-70 and B...
Induction of Specific Cytotoxic Lymphocytes in Mice Vaccinated with Brucella abortus RB51
Infection and Immunity, 2001
A safe, more sensitive, nonradioactive, neutral red uptake assay was adopted to replace the traditional 51 Cr release assay for detection of Brucella-specific cytotoxic T lymphocyte (CTL) activity. Our studies indicated that Brucella abortus strain RB51 vaccination of mice induced specific CTLs against both strain RB51-and strain 2308-infected J774.A1 macrophages but not against Listeria monocytogenes-infected J774.A1 cells. The antigenspecific cytotoxic activity was exerted by T lymphocytes but not by NK cells. CD3 ؉ CD4 ؉ T cells secreted the highest level of gamma interferon (IFN-␥) and were able to exert a low but significant level of specific lysis of Brucella-infected macrophages. They also exerted a low level of nonspecific lysis of noninfected macrophages. In contrast, CD3 ؉ CD8 ؉ T cells secreted low levels of IFN-␥ but demonstrated high levels of specific lysis of Brucella-infected macrophages with no nonspecific lysis. These findings indicate that B. abortus strain RB51 vaccination of mice induces specific CTLs and suggest that CD3 ؉ CD4 ؉ and CD3 ؉ CD8 ؉ T cells play a synergistic role in the anti-Brucella activity.
Infection and Immunity
In contrast to immunity against some other facultative intracellular parasites, protective immunity against Brucella abortus is mediated in mice by antibodies as well as by cell-mediated immune responses. It was the purpose of this study to determine whether antibody alone would prevent infection with B. abortus. The majority (82%) of CD-1 outbred mice infected with 100 CFU of virulent B. abortus 2308 preincubated with graded quantities of an O polysaccharide-specific IgG2a monoclonal antibody (MAb) were free of infection 1. 2, 4, and 6 weeks later, based on detection limits of 13 brucellae per spleen and 39 per liver. Infection was present in 95% of control animals. Similar results were obtained with a challenge dose of 500 CFU, but with a challenge dose of 5,000 CFU, infection became established even with the highest concentration of MAb used (50 micrograms of MAb per 5,000 brucellae). Pretreatment with an O polysaccharide-specific IgG1 MAb or with convalescent-phase serum diminis...
Comparative Immunology, Microbiology and Infectious Diseases, 1979
Abstraet--A Brucella abortus soluble antigen (BASA) preparation and fractions obtained thereof, by column chromatography, were compared in terms of their ability to induce specific lymphocyte stimulation responses (LSR) in lymphocytes from cattle infected with B. abortus. Endotoxin and protein contents and the fractions were determined. The LSR induced by BASA and the fractions were compared in terms of correctly identifying samples from infected and non-infected cattle. The sensitivity and specificity for each preparation were determined and these two attributes were then correlated with endotoxin and protein content. The results suggest that lymphocyte stimulation had greater association with relative protein content than endotoxin content of the antigen preparations. Key words: Brucella abortus soluble antigen (BASA), lymphocyte stimulation responses (LSR), endotoxili contents, protein contents CERTAINES REACTIONS D'IMMUNOSTIMULATION LYMPHOCYTAIRE IN VITRO DES FRACTIONS BR UCELLA ABOR TUS OBTENUES PAR C HROMATOGRAPHIE SUR C OLONNES Resum~----Un antigone soluble de Brucella abortus, ainsi que des fractions obtenues par chromatographie sur eolonne, ont 6t6 compar6s quanta leur capacite d'induire une r6ponse pro-lif6rative sp/:cifique de lymphocytes provenant de bovins infect6s par B. abortus. Les quantit6s d'endotoxine et cLe prot6ines contenues darts l'antig~ne soluble et dans les fractions ont ete determin6es. Les stimulations lymphocytaires induites par l'antig/me soluble ou les diff6rentes fractions ont et6 compar~es quant /l leur valeur diagnostique sur des 6chantillons de sang provenant d'animaux infectes et non infect6s. La sensibilit6 et la sp6cificit~ de chaque pr6paration ont ~te 6tudi6es et la relation entre ces deux facteurs et le contenu en endotoxine et en proteines a et6 recherch6e. Les r6sultats obtenus sugg~rent que la stimulation lymphocytaire est plus fonction du contenu des pr6 parations en proteines qu'en endotoxine. Mots-clefs: Antigene soluble de Brucella abortus (ASBA), r6actions de stimulation lymphocytaire (RSL), constituants d'endotoxine, constituants prot~iques * ISCO Golden Retriever (model 820), Instrumentation Specialties Co., Lincoln, Nebras., U.S.A. ~: Arthur H. Thomas, Co., Philadelphia, Pen., U.S.A.
Veterinary Microbiology, 1991
A study was conducted to determine whether the protection induced in mice by a primary inoculation of lipopolysaccharide from Brucella abortus would be enhanced by a second inoculation given at different time intervals. Protection was challenged by exposure of the mice to a virulent culture of B. abortus strain 2308. Reduced mean viable count and/or splenic weights were the criteria of protection. There was no significant difference (P> 0.05) in the protective responses among mice given a single inoculation. Vaccinated mice were significantly (P<0.05) better protected than were nonvaccinated mice. Mice given vaccinal inoculations simultaneous with challenge exposure were less protected (P < 0.001) than were mice vaccinated prior to challenge, but were better protected (P < 0.010) than were nonvaccinated mice.
Immune response strategies of Brucella melitensis and their antigens in rats
Iraqi Journal of Veterinary Sciences
Brucella melitensis is an intracellular bacterium and is the main brucella species that cause abortion and placenta retention in sheep and goats. It has many mechanisms to evade the immune response. The current study aimed to investigate Brucella melitensis strategies for producing immune responses in rats after challenging the bacterium. For this purpose, live and killed Brucella melitensis REV1 strain was given to rats subcutaneously, and immunological markers like TLR2, TLR4, IFN-γ, and anti-brucella antibodies were determined. The results showed that the level of immunological markers like TLR2 and TLR4 did not significantly increase in rat groups inoculated with live Brucella melitensis, while it increased in the rats' groups vaccinated with the sonicated Brucella melitensis; also, the results showed an increase in the level of IFN-γ and antibrucella antibody titers in all animal groups. The study concluded that the inoculation with killed bacteria and REV1 could protect the animals against challenging doses, as seen when the groups were inoculated with challenge dose of the bacterium.
Immunotoxic effect of thiamethoxam in immunized mice with Brucella abortus cultural filtrate antigen
Veterinary World, 2016
This study was planned for determination the toxic effect of thiamethoxam (TMX) in immunized mice with Brucella abortus culture filtrate antigen (CFBAgs) (as a vaccine) and its role of TMX on decrease activity of B. abortus antigen on eliciting of humoral and cellular immunity. Materials and Methods: To achieve these goals 60 female mice were used, 7-8 weeks age, they were divided equally into three groups (20 in each group) and treated as follows: 1 st group: Mice were immunized with CFBAgs intraperitoneally in two doses, 2 weeks intervals with (protein concentration 2 mg\ml), 2 nd group: Mice immunized as in the 1 st group and was administrated orally with 1/10 lethal dose 50% of TMX (83.7 mg/kg B.W.) for 4 weeks daily, 3 rd group was administrated orally with 0.3 ml normal saline served as a control group. At day 28 post immunization (PI) delayed type hypersensitivity (skin test) was done, and serum samples were collected at day 30 (PI) for detection of passive hemagglutination test (PHA); interferon gamma (IFN-γ) which was done by enzyme-linked immunosorbent assay test in addition to phagocytes assay. Results: The results of skin test post injection with soluble antigen of B. abortus intradermally showed a high significantly mean values at p≤0.05 of footpad skin thickness in the 1 st group of mice which recorded (0.51±0.002 mm) as compared with the 2 nd group of mice which showed (0.08±0.002 mm) after 24 h; the mean values of skin thickness were declined in the 1 st mice (0.46±0.002) and 2 nd mice (0.070±0.001) at 48 h; control group showed a negative results. These results were agreed with results of serum levels of IFN-γ (pg/ml) that showed that a significant increase the vaccinated 1 st group (406.36±1.52), than those values in the 2 nd group (151.61±0.89) and negative result in 3 rd group (46.47±0.60), in addition to results of PHA test which showed a significant increase in antibody titer in the 1 st group (139±12.16) with low level of serum antibody in the 2 nd group (7.66±0.33). Phagocytic ratio results in the 1 st group showed an increase to reach (18.55±0.44) than a ratio in the 2 nd group (13.24±0.32) and the control group (5.46±0.25). Conclusion: It was concluded that TMX induced suppression of humoral and cellular immune responses in immunized mice with CFBAgs.
Characteristics of the immune response during acute brucellosis in Sprague-Dawley rats
The Journal of Infection in Developing Countries, 2009
Background: Brucella is a facultative, intracellular pathogen that causes severe disease in animals and humans. Immunity against Brucella involves both humoral and cellular responses. To investigate the characteristics of immune response in acute brucellosis in Sprague-Dawley (SD) rats, IgG and its subclass specific immunoglobulins' (IgG1 and IgG2a) response in sera against B. abortus biotype 1 infection were studied. Methodology: Thirty-six rats were inoculated intraperitoneally with 0.1 ml apyrogenic saline containing 1 × 10 10 colony forming unit (CFU) of B. abortus biotype 1 Korean bovine isolate. Four rats were used as uninfected controls. The sera were collected from infected rats at 3, 7, 14, 21, 28, 35, 42, 49, and 56 days post infection (DPI) and screened for Brucella specific antibody response by the rose bengal plate test (RBPT). IgG and its subclass specific immunoglobulins' (IgG1 and IgG2a) response in the sera were measured by a lipopolysaccharide (LPS) based indirect enzyme-linked immunosorbent assay (IELISA). Results: Brucella specific IgG, IgG1 and IgG2a responses in the sera of infected rats were detected from 3 DPI by IELISA. IgG and IgG1 concentrations in sera reached the peak level at 35 DPI, and then the concentrations gradually declined to the end of the experiment. IgG2a concentrations in the sera remained almost constant from 7 DPI until the end of this study. Conclusion: In acute brucellosis, IgG2a response (indicative of a Th1 response) was found to be significantly dominant over IgG1 response (indicative of Th2 response) (P < 0.001).
Humoral Immunity in mice Mediated by Monoclonal Antibodies Against the A and M Antigens of Brucella
Journal of Medical Microbiology, 1989
All smooth strains of Brucella bear two lipopolysaccharide (LPS) antigens in a ratio that defines the classification of strains in serovars, A (A > M), M (M >A) and A.M (A = M). Anti-LPS-A monoclonal antibodies (MAb-A) were previously shown to convey protection to mice against B. abortus (A) strain 544, as shown by lower spleen counts than in controls at days 7 and 21 after challenge. Anti-LPS-M monoclonal antibodies (MAb-M) were obtained and tested for M-specificity with LPS from reference strains by ELISA, by agglutination of LPS-coated latex particles, and by inhibition of this agglutination. Antigens A and M of three strains were quantified by a homologous LPS-latex and MAb agglutination inhibition assay. Protection conferred by MAb-A and MAb-M against three strains, B. abortus 544 (A), B. abortus 292 (M) and B. melitensis H38 (M)
Characterization of Brucella abortus Soluble Antigen Employed in Immunoassay
Journal of Clinical Microbiology, 1980
A soluble antigen extract of Brucella abortus (BASA) has been prepared by the National Veterinary Services Laboratories and furnished to a number of workers who are examining antibody-mediated and cell-mediated immune responses of cattle infected with B. abortus. Three lots of BASA were examined. There were quantitative but not qualitative differences among lots by content of protein, total carbohydrate, hexose, fatty acid, and 2-keto-3-deoxyoctonic acid. The presence of smooth lipopolysaccharide was demonstrated by the presence of 2-keto-3-deoxyoctonic acid and lipid, by Limulus lysate gelation activity, and by formation of characteristic lipopolysaccharide precipitates in immunoelectrophoresis. A polysaccharide antigen as well as two nonsurface antigens, A2 and C, were also identified. BASA is a satisfactory antigen for use in the enzyme-linked immunosorbent assay since the smooth lipopolysaccharide component bound to polystyrene and functioned in the test. Normal murine spleen ce...