[Checking of anti-Brucella vaccines by counting the Brucella in the spleen of intraperitoneally innoculated, vaccinated or unvaccinated, mice].
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A good survival of brucellae in the organism with the development of a stable and prolonged immunity was noted in vaccination of guinea pigs with live brucella vaccine prepared from the Br. abortus strain 19-BA. Local reactions were weak, and no general reactions were observed. In using the jet method of vaccination with the injectors bpi-4 and bi-2 on a limited group of persons (194) the local and general untoward response was weak, but in immunological respect this method proved to be effective when the vaccine was administered in a dose of 2.5 x 10(8) of microbial cells. Jet method can be recommended for express vaccination against brucellosis.
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Immunity assays in guinea-pigs based on spleen mass: body mass ratios showed that live vaccines were markedly more effective than inactivated oil adjuvant vaccines in providing protection against challenge with 5 000 virulent organisms.
Cell-mediated immune (CMI) responses in cattle vaccinated with Brucella abortus strain 19 vaccine during calfhood were studied by an in vitro lymphocyte stimulation assay. Cattle were grouped in six groups according to the age after vaccination, and CMI responses of these groups, as well as of individual animals, were compared. Lymphocytes were prepared from peripheral blood of these cattle by the Ficoll-diatrizoate technique. Lymphocytes were then cultured with B abortus-soluble antigen. Culture results were assayed for [3H]thymidine incorporation into DNA by liquid scintillation spectrometry. On a group basis, B abortus-soluble antigen induced lymphocyte stimulation responses in lymphocytes from all the groups, except the sixth group which contained animals that had been vaccinated the longest time (18 to 24 months before this experiment). Animals that had been vaccinated for 3 to 6 months had the highest lymphocyte stimulation response. Seroagglutination tests were conducted simultaneously with the lymphocyte stimulation test, but there was no apparent correlation between the concentrations of humoral antibodies and the CMI responses as measured by in vitro specific lymphocyte stimulation. The lymphocyte stimulation test exhibited significantly higher specificity (P less than 0.005) than the serologic tests.
Since age-dependent diminished macrophage function has been related to the increased susceptibility of newborn mice to herpes simplex virus (HSV) infection, the effect of several agents which might activate macrophages or other cells involved in immunity was investigated. BCG, typhoid vaccine, brucella vaccine, levamisole, or staphage lysate were administered to newborn mice prior to challenge with HSV-2. Of these agents, only BCG, administered ip or id 6 days prior to challenge, was found to increase the survival rate of newborn mice. The possible use of BCG, alone or in combination with other modalities, for the prevention or treatment of neonatal HSV infections is discussed.
Cell-mediated immune responses in cattle naturally infected with strains of Brucella abortus and in cattle vaccinated with B abortus strain 19 during calfhood were studied by an in vitro lymphocyte-stimulation procedure. Lymphocytes were prepared from peripheral bovine blood by the Ficoll-diatrizoate technique, suspended in RPMI-1640 medium (1.5 X 10(6) lymphocytes/ml), cultured with B abortus-soluble antigen or phytohemagglutinin, and incubated for 6 days. Sixteen hours prior to termination of incubation, cultures were labeled with 1 muCi of [3H]thymidine (3HdT) and, after harvesting, assayed for 3HdT incorporation into DNA by liquid scintillation spectrometry. Lymphocytes from cattle with bacteriologically confirmed isolation of B abortus underwent a significantly higher lymphocyte stimulation with B abortus-soluble antigen than did cattle vaccinated with B abortus strain 19 during calfhood (P less than 0.005). Standard seroagglutination tests were conducted simultaneously with lymphocyte-stimulation tests, but there was no apparent correlation between levels of humoral antibodies and the cell-mediated immune responses as measured by in vitro specific lymphocyte stimulation.
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Cell-mediated immune responses in cattle adult-vaccinated with Brucella abortus strain 19, cattle infected with B abortus field strain, and nonexposed cattle were studied by an in vitro lumphocyte-stimulation test (LST). Lymphocytes were prepared from peripheral bovine blood by the Ficoll-diatrizoate technique, and results were assayed for [3H]thymidine incorporation into DNA by liquid scintillation spectrometry. Serotests and bacteriologic isolation attempts were conducted simultaneously with LST. Lymphocytes from cattle infected with field strains had significantly (P = 0.01) higher specific lymphocyte-stimulation inexposed controls. The LST, the serum standard-tube agglutination test (STT), the Rivanol (RIV) test, and the complement-fixation (CF) test correctly classified cattle from which field strains and strain 19 of B abortus were isolated. The LST was negative in cattle vaccinated with B abortus strain 19 (nonshedding), but the three serotests had many false-positive reactions. The CF test had the least false-positive reaction, followed by the RIV test, and the STT was the least specific. Well before the three serotests became positive, the LST was positive in samples from some cattle during the incubation period of the infection. There was little or no correlation between cell-mediated immune responses (as measured by LST) and serum antibody responses (as measured by STT, RIV test, and CF test) in vaccinated but culture-negative cattle and in some nonvaccinated cattle during the incubation period.
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The serological response of chicks to Brucella abortus strain 19 was monitored over a period of seven weeks to assess the degree of immunosuppression caused by vaccination at one day of age with two infectious bursal disease vaccines. One of the vacy. This vaccine caused severe immunosuppression judged by the minimal serological response following B abortus inoculation. The test also detected a significant delay caused by the other vaccine in the development of the serological response but the maximum titre was not significantly different from that in chicks which had received no infectious bursal disease vaccine.
Chronic protein-deficiency in weanling mice caused variable suppression of the humoral plaque-forming cell (PFC) responses to sheep erythrocytes. This was most prominent at high antigen doses and did not increase when mice were maintained on the diets for longer periods. Antibody responses produced by deficient mice were often short-lived and involved high levels of IgM. Total PFC counts were depressed slightly more than were circulating antibodies. Antibody responses to Brucella abortus were slightly decreased by protein-deficiency at high antigen doses but were normal or elevated at lower doses, the proportion of IgM produced was increased and the splenomegaly response to B. abortus was severely depressed. These results suggest that the depression of antibody production by protein-deficiency is not simply due to an impairment of helper T cell function, but a reduction in the availability or effectiveness of macrophage and regulatory or suppressor T cells may be important.
Cell-mediated immunity to Brucella abortus S19 vaccine was measured in young heifers by the microassay for stimulation of protein synthesis (SPS) with [3H]leucine and the skin test for delayed hypersensitivity. Brucella melitensis protein allergen and a crude B abortus S19-soluble antigen were compared in the SPS test. The SPS test was negative in 5 unvaccinated heifers and strongly positive in 3 twice-vaccinated steers. However, the SPS test was positive only in 13 of 30 S19-vaccinated heifers and the delayed hypersensitivity in 9 of 29 S19-vaccinated heifers. The 2 tests gave good agreement. Vaccination-induced residual antibody titers were partly correlated with the outcome of the tests used to measure cell-mediated immunity.