[Tuberculosis--still a current problem].
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Africa↗
Biomedical subjects
J Bennedsen
Publications and source records attributed to J Bennedsen.
Identification of immunodominant antigens during infection with Mycobacterium tuberculosis.
T lymphocytes isolated from mice infected with Mycobacterium tuberculosis response vigorously to proteins secreted by the bacilli and these antigens may be of importance in the generation of protective immunity against the disease. In this study, short-term culture filtrate (ST-CF), which constitutes a complex mixture of secreted proteins, was fractionated by a modified preparative SDS-PAGE technique. The ability of each fraction to be recognized by T cells isolated from infected mice was evaluated by quantifying proliferation and IFN-gamma production in cell cultures. Two molecular mass regions 4-11 and 26-35 kDa were found to possess marked stimulatory properties. Four potent single antigens were mapped within the stimulatory regions. These purified antigens stimulated T cells isolated from mice at the height of a tuberculous infection to produce large amounts of IFN-gamma. Two of these stimulatory antigens belonged to the antigen 85 complex.
Analysis of Variance↗
Detection of mycobacteria from blood and bone marrow: a decade of experience.
This study reports our experience with methods used at our department from 1981 through 1990 for detection of mycobacteria in blood and bone marrow specimens. Direct inoculation on Lowenstein Jensen media was replaced by Isolator lysis-centrifugation followed by inoculation on conventional solid media, and the Bactec 12B and Bactec 13A systems. A total of 3033 specimens were analyzed. A total of 137 mycobacterial isolates were obtained from 42 patients, all HIV-positive except one. Mycobacteremia caused by M. avium-intracellulare (83%), M. tuberculosis, M. scrofulaceum and M. kansasii was found. Of 680 blood specimens tested by the last three methods, 7.6% were found to be positive by at least one method and revealed recovery rates of 6.8% for the Isolator-solid media system, 3.4% for the Isolator-12B system and 6.9% for the 13A system (all isolates MOTT). Mean detection times for 21 cultures found positive by all three methods were 23.6, 23.3 and 17.7 days for the Isolator-solid media, Isolator-12B and 13A systems, respectively, with a significantly shorter detection time for the 13A system. Low degree (less than 1 cfu/ml) mycobacteremia (MOTT) caused delay in the Isolator-solid media and the 13A systems and no detection in the Isolator-12B system. Antituberculous therapy significantly prolonged the detection times for MOTT in the 13A system in contrast to the other systems.
Bacteremia↗
Results of the third immunology of leprosy/immunology of tuberculosis antimycobacterial monoclonal antibody workshop.
An international workshop was sponsored by the World Health organization to screen new antimycobacterial monoclonal antibodies and to identify antibodies which could be recommended as standard reagents giving consistent results under differing assay conditions. Fifty-eight antibodies were submitted to the workshop by eight independent laboratories. Nineteen of the antibodies recognized antigens distinct from those identified in earlier workshops, defining at least 10 new protein antigens. Monoclonal antibodies characterized in the workshop provide a set of convenient reagents for further characterization of mycobacterial antigens.
Animals↗
Infection with Mycobacterium bovis in a patient with AIDS: a late complication of BCG vaccination.
We present a 28-year-old HIV-infected man with a 2-year delayed complication of BCG immunization. When immunized the man was healthy, with an unknown HIV status, but 2 years later he was diagnosed with AIDS because of a Pneumocystis carinii pneumonia. He was successfully treated and discharged in a state of good health. A few months later he presented with an enlarged lymph node and Mycobacterium bovis, BCG strain, was cultured. No sign of dissemination was found. We discuss the indications for BCG vaccination in adults, especially in areas and in populations with a high prevalence of HIV.
Acquired Immunodeficiency Syndrome↗
Proteins released from Mycobacterium tuberculosis during growth.
Proteins secreted from Mycobacterium tuberculosis during growth are believed to be important for protective immunity against tuberculosis. We have investigated the growth of M. tuberculosis in an enriched liquid medium. The release of isocitrate dehydrogenase from the bacilli served as a marker of autolysis and was observed during the late logarithmic growth phase. The release of proteins during the culture period was investigated by enzyme-linked immunosorbent assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Three major groups of proteins, which differed markedly with respect to profile of release and location in intact bacilli, were defined. A short-term filtrate devoid of autolytic products was defined and found to be composed of 33 major components. Five proteins were identified by monoclonal antibodies. Pronounced superoxide dismutase activity was detected in the filtrate. The enzyme was purified and identified as a dominating component of short-term filtrate.
Animals↗
Quantitative and qualitative studies on the major extracellular antigen of Mycobacterium tuberculosis H37Rv and Mycobacterium bovis BCG.
The Mycobacterium tuberculosis antigen 85 is a biologically important antigen. Tuberculosis patients may have strong antibodies against it, and their peripheral blood mononuclear cells respond to it with gamma-interferon production and lymphocyte proliferation. Antigen 85 is actively secreted into the culture medium during culture in vitro and is known to bind human fibronectin. A double-antibody enzyme-linked immunosorbent assay (ELISA) for quantification of antigen 85 is described. A mouse monoclonal antibody, HYT27, was used as capture antibody in the assay. HYT27 was characterized in crossed immunoelectrophoresis and found to bind all three components of the antigen 85 complex. By radioimmunoassay, HYT27 was found to bind equally well to antigens 85A and 85B. In the ELISA assay, a rabbit anti-antigen 85 antiserum was used in the second antibody layer. The specificity of the assay was tested using several different antigen preparations. The purified BCG 85A and 85B components were compared, and there was a 10 times lower sensitivity for antigen 85A due to weaker rabbit antibodies toward this component. The purified components MPT44 and MPT59 from M. tuberculosis H37Rv were compared with the components of BCG and found to correspond to BCG 85A and 85B, respectively. Mycobacterium kansasii and Mycobacterium avium both contained partially identical antigens. Small amounts of antigen 85 were detected in Mycobacterium leprae sonicates. Detecting antigen 85 by sensitive methods may be of great value in the early diagnosis of mycobacterial disease.
Antibodies, Monoclonal↗
Affinity purification, biological characterization and serological evaluation of defined antigens from Mycobacterium tuberculosis.
Nine mycobacterial antigens have been purified by affinity chromatography using monoclonal antibodies (m. abs) as immunosorbent. The importance of the individual antigens have been evaluated by different immunological methods.
Animals↗
Interspecies reactivity of five monoclonal antibodies to Mycobacterium tuberculosis as examined by immunoblotting and enzyme-linked immunosorbent assay.
Five different murine monoclonal antibodies (MAbs) to Mycobacterium tuberculosis were examined for degree of cross-reactivity with other mycobacterial species by enzyme-linked immunosorbent assay and immunoblotting. One MAb reacted solely with M. tuberculosis and M. bovis BCG. Two of the MAbs reacted with all mycobacterial species examined, whereas two MAbs demonstrated a limited reactivity pattern. The epitopes are located on molecules susceptible to protease treatment, and two of these molecules possess concanavalin A-binding moieties. Two of the antigens defined by these five MAbs are present in tuberculin purified protein derivative.
Animals↗
Activation of macrophages assessed by in vivo and in vitro tests.
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Animals↗
Correlation between in vivo and in vitro functional tests for activated macrophages.
Mice undergoing graft-versus-host reaction, skin grafting, and inoculation with tumor cells were tested for nonspecific resistance by intravenous challenge with Listeria monocytogenes. Peritoneal exudate macrophages from mice treated in a similar manner were tested in vitro for increased degradation of [1-14C]glucose, ability to degrade antigen/antibody complexes, ability to inhibit intracellular growth of listeria, and staining for beta-galactosidase. There was good correlation between in vivo resistance towards L. monocytogenes and in vitro inhibition of intracellular growth. There was also good correlation between increase in beta-galactosidase and in vivo resistance in mice undergoing a graft-versus-host-reaction.
Animals↗
In vitro effect of mitogen-activated speen cells and their culture supernatants on mouse peritoneal macrophages as assayed by intracellular killing of Listeria monocytogenes and content of beta-galactosidase.
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Animals↗
Macrophage activation by lymphokines and after direct contact with sensitized lymphocytes: histocompatibility requirements and the effect of inhibitors.
A comparison was made between two procedures which give rise to in vitro activation of normal mouse peritoneal macrophages: (1) normal macrophages were incubated for 22 h with sensitized lymphocytes and antigen (assay A), and (2) normal macrophages were incubated for 70 h with supernatants from sensitized lymphocytes and antigen (assay B). The activation of macrophages was measured as an increase in 1-14 C glucose oxidation. Combinations of lymphocytes and macrophages is probably necessary in assay A.s from different mouse strains demonstrated that activation of macrophages in assay A, but not in assay B, required cells which were derived from strains of mice sharing identical H-2 antigens. Treatment of lymphocytes with mitomycin C blocked the activation of macrophages in both assays. Addition of alpha-L-fucose (0.1 M) during the incubation period blocked the activation of macrophages in assay B, but not in assay A. It is concluded that there is a qualitative difference between the two methods for activating macrophages and that direct contact between lymphocytes and macrophages is probably necessary in assay A.
Animals↗
In vitro studies on normal, stimulated and immunologically activated mouse macrophages. I. Oxidation of 1-14C glucose by macrophages in monolayer cultures.
Oxidation of 1-14C glucose of BCG/listeria activated, proteose-peptone stimulated and typhoid-parathypoid vaccine (T.A.B.) stimulated peritoneal and spleen macrophages from C3H, CBA and C57Bl mice was determined at various times after treatment. The study showed no major differences between the oxidation of stimulated and activated macrophages, but a variation was seen in C3H mice in the kinetics of glucose oxidation of BCG/listeria activated and proteose-peptone stimulated peritoneal macrophages. It was also observed, in these mice, that the response appeared and disappeared earlier in the spleen than in the peritoneal macrophages.
Animals↗
In vitro studies on normal, stimulated and immunologically activated mouse macrophages. II. Degradation of radioactive antigen/antibody complexes.
Radioactive antigen/antibody complexes are degraded to a greater extent by PE macrophages from C3H, CBA and C57Bl mice injected intraperitoneally with proteose-peptone (stimulated macrophages) and T.A.B. vaccine than by PE macrophages from normal mice and mice immunized with BCG (activated macrophages). On the other hand, spleen macrophages from mice immunized with BCG had a greater digestive capacity than macrophages from mice injected with proteose-peptone or T.A.B. vaccine or from normal mice. Thus, activated macrophages are capable of degrading antigen/antibody complexes, provided that the cells are harvested from one of the foci of BCG infection.
Animals↗
In vitro studies on normal, stimulated and immunologically activated mouse macrophages. III. Intracellular multiplication of Listeria monocytogenes.
The intracellular generation time of Listeria monocytogenes is increased in monolayers of PE macrophages from mice immunized with BCG, listeria and T.A.B. vaccine as compared with monolayers of PE macrophages from normal mice stimulated intraperitoneally with proteosepeptone. Thus non-specifically stimulated and immunologically activated macrophages, although sharing several other characteristics, differ in their capacity to control intracellular bacterial infection.
Animals↗