Hospital medical staffs: the future is now.
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Contract Services↗
Biomedical subjects
T M Daniel
Publications and source records attributed to T M Daniel.
Evaluation of mycobacterial antigens in an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of tuberculosis.
Five mycobacterial antigens were compared in an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of tuberculosis. The antigens studied were an unheated sterile culture filtrate of Mycobacterium tuberculosis, tuberculin purified protein derivative (PPD) from M. tuberculosis (PPDa), purified cytoplasmic protein antigens 5 and 6 from M. tuberculosis, and a PPD prepared from M. kansasi (PPDk). Multivariate analysis of variance showed that geometric mean titres obtained with each of the antigens in ELISA were significantly different in tuberculosis patients and in control groups. The covariation of the ELISA results with the five antigens was highly interdependent. Analysis of receiver operating characteristics revealed that the most accurate test was obtained with antigen 5. M. tuberculosis PPD, M. tuberculosis antigen 6, and M. tuberculosis culture filtrate were, in descending order, less accurate.
Antibodies, Bacterial↗
Specificity of Mycobacterium tuberculosis antigen 5 determined with mouse monoclonal antibodies.
Four mouse monoclonal antibodies have been developed which react with Mycobacterium tuberculosis antigen 5. Each of these monoclonal antibodies has been used to prepare immunoabsorbents, and antigen has been isolated from unheated M. tuberculosis H37Ra culture filtrate by affinity chromatography with these absorbents. Antigen thus obtained was found to be similar in tuberculin reactivity and in enzyme-linked immunosorbent assays to antigen 5 isolated from polyclonal immunoabsorbents, and the protein yields from the monoclonal immunoabsorbents were similar to those from polyclonal absorbents. Antigen recovered from monoclonal absorbents cross-reacted with antigens of Mycobacterium kansasii in delayed skin tests. Immunoelectrophoresis demonstrated that the major component of the antigens eluted from the monoclonal immunoabsorbents was mycobacterial arabinomannan. Antigen 5 was not identified in the eluates by immunoelectrophoresis. These observations support the hypothesis that antigen 5 and M. tuberculosis arabinomannan contain a single major shared epitope.
Animals↗
Serodiagnosis of pulmonary tuberculosis in Argentina by enzyme-linked immunosorbent assay (ELISA) of IgG antibody to Mycobacterium tuberculosis antigen 5 and tuberculin purified protein derivative.
IgG antibody to Mycobacterium tuberculosis antigen 5 and tuberculin purified protein derivative (PPD) was measured, by enzyme-linked immunosorbent assay (ELISA), in serum samples from 86 patients with active pulmonary tuberculosis and 91 non-tuberculous control subjects from Santa Fé, Argentina. The geometric mean titre for the tuberculosis patients was 74.6 with antigen 5 and 99.5 with PPD. In 91 control subjects the geometric mean titres were 3.6 and 15.6 respectively. Titres were not related to tuberculin reactor status or prior BCG vaccination. At a serum dilution end-point of 1:40, ELISA with antigen 5 had a sensitivity of 81.4% and a specificity of 93.4% for tuberculosis. At 1:40, ELISA with PPD showed a sensitivity of 82.6% and a specificity of 54.9% for tuberculosis. Applied at a serum dilution of 1:40 to a hypothetical model population with a tuberculosis prevalence of 2%, ELISA using antigen 5 would correctly classify 93.2% of persons and ELISA with PPD, 55.5%. At a dilution of 1:80, accuracy is increased to 99.3% with antigen 5 and 83.3% with PPD, but sensitivity decreases to 64.0% with antigen 5 and 72.1% with PPD. Thus, antigen 5 is more accurate than PPD for the diagnosis of tuberculosis using ELISA.
Antigens, Bacterial↗
Immunochemical analysis of tuberculin purified protein derivative with special reference to United States-Japan antigen 7.
Immunochemical analysis of tuberculin purified protein derivative (PPD) by crossed immunoelectrophoresis and other agar gel techniques revealed substantial variation in antigen levels among PPDs and significant differences that were dependent upon the antiserum used. D-Arabino-D-mannan was identified as a major polysaccharide constituent of PPD; D-arabino-D-galactan and United States-Japan antigen 6 were also present. United States-Japan antigen 7 was found to be a major anodal constituent of all preparations of PPD. This antigen was purified and demonstrated to be a molecularly heterogenous small protein or large peptide with an average molecular weight of approximately 8,000. Antigen 7 possessed tuberculin activity with about one fourth the potency of PPD. When studied in animals sensitized with Mycobacterium tuberculosis and Mycobacterium kansasii, antigen 7 displayed no more tuberculin specificity than PPD.
Chromatography, Affinity↗
Isolation, characterization, and specificity of a glycoprotein antigen from Mycobacterium kansasii.
We have isolated in highly purified form and characterized a glycoprotein antigen from culture filtrates of Mycobacterium kansasii. Immunoelectrophoretic studies demonstrated that this antigen is present only in M. kansasii among 14 species of mycobacteria studied. It is a potent tuberculin skin test antigen, but skin test reactions to it in sensitized guinea pigs display only limited specificity. Enzyme-linked immunoabsorbent antibody assays of serum samples from patients suffering from diseases caused by M. kansasii, M. intracellulare, and M. tuberculosis display no specificity with this antigen. These findings are best explained by hypothesizing a single highly antigenic determinant that is present as a moiety on many mycobacterial antigens and is widely shared among mycobacteria.
Animals↗
Serologic diagnosis of bone and joint tuberculosis by an enzyme-linked immunosorbent assay.
Sera from patients receiving treatment for active bone and joint tuberculosis and sera from patients with inactive bone and joint tuberculosis were examined by an enzyme-linked immunosorbent assay for antibody to antigen 6, a homogeneous protein prepared from Mycobacterium tuberculosis strain H37Ra by immunosorbent affinity chromatography. Sera from 21 control subjects had a geometric mean titer of 1:6 with no difference between tuberculin purified protein derivative-positive and -negative patients. Sera from 20 patients with inactive disease had a geometric mean titer of 1:19. Fifteen patients receiving treatment for M. tuberculosis infection had a geometric mean titer of 1:179, which is significantly different from the geometric mean titers of both of the patients with inactive tuberculosis (P less than 0.001) and the control subjects (P less than 0.001). At a cut-off titer of 1:32, the sensitivity of the assay is 94% and the specificity for the control subjects and patients with inactive disease was 100%.
Adolescent↗
Robert Koch, tuberculosis, and the subsequent history of medicine.
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Bacteriology↗
The tuberculin specificity in humans of Mycobacterium tuberculosis antigen 5.
Mycobacterium tuberculosis antigen 5 is a protein antigen limited in distribution to M. tuberculosis and M. bovis and capable of eliciting typical delayed tuberculin skin test reactions in humans. A single large batch of this antigen was purified by immunoabsorbent affinity chromatography and used to skin test patients with tuberculosis and other mycobacterial infections and healthy persons in general populations in geographic areas where nonspecific tuberculin reactivity is frequently encountered. Antigen 5 was found to be no more specific as a tuberculin antigen than PPD. If the available data are accepted, then either a disparity in antigen recognition by antibody and T lymphocytes may exist or the widely accepted hypothesis attributing nonspecific tuberculin reactivity to antigenic cross reactivity with other mycobacteria may be incorrect.
Adult↗
Serodiagnosis of tuberculosis using the enzyme-linked immunoabsorbent assay (ELISA) of antibody to Mycobacterium tuberculosis antigen 5.
The serologic response to purified mycobacterial antigen 5 was examined using an enzyme-linked immunoabsorbent assay in 75 patients with pulmonary tuberculosis and 150 control subjects. Two patient and 6 control groups were studied. The serums from the patient groups had significantly higher IgG antibody concentrations than infected control subjects with good specificity. In patients living in low prevalence areas where the results of skin tests in the majority of the population are negative, an antibody titer greater than or equal to 1:40 would have a 95.8% specificity for active disease. For patients living in a high prevalence area where the results of skin tests in the majority of the population are positive, then an antibody titer greater than or equal to 1:40 would have a 79.9% specificity for active disease. Serums from patients with active nontuberculous mycobacterial infection had intermediate mean titers.
Antibodies, Bacterial↗
Antigranulocyte antibodies and deranged immune function associated with phenytoin-induced serum sickness.
A phenytoin-associated serum-sickness-like syndrome was followed by severe pancytopenia in a 17-year-old female. Serum obtained during the acute phase of her illness contained antibodies against homologous eosinophils and neutrophils, antibodies against autologous neutrophils, and antibodies to phenytoin. The serum also inhibited bone marrow granulocyte colon formation in vitro. Lymphocyte transformation by pokeweed mitogen was impaired and, after recovery, the patient's lymphocytes were transformed in vitro by the addition of phenytoin. These data demonstrate that profound derangements of immunity can be associated with a phenytoin-induced serum-sickness-like illness.
Adolescent↗
Persistence of protein, carbohydrate and was components of tubercle bacilli in dermal BCG lesions.
Dermal tuberculous lesions were produced in rabbits with Bacille Calmette Guèrin and biopsied on days 4, 14, 21, 35, and 56. Frozen sections prepared from the biopsy specimens were stained by the peroxidase-antiperoxidase immunocytochemical technique with antisera against whole tubercle bacilli and antisera against the protein B, polysaccharide I, and wax D fractions of tubercle bacilli. In the primary lesions, protein B was often undetectable at 21 days, and polysaccharide I was often undetectable at 35 days. Wax D disappeared more slowly than the other bacillary components evaluated, and some wax D was still found at 56 days (if the lesions had not completely healed). Antisera against intact tubercle bacilli produced results similar to those produced by antiserum against wax D. These studies suggest that the chronicity of tuberculous lesions is due, at least in part, to the persistence of the wax D-like component of tubercle bacilli.
Animals↗
Initial clinical trial of Mycobacterium tuberculosis antigen 5 in tuberculin-positive human subjects.
A single lot of Mycobacterium tuberculosis antigen 5 was used in graded doses to skin test 92 persons in a geographic area where nonspecific tuberculin reactivity is uncommon. In 47 tuberculin-positive subjects, antigen 5 was found to have a potency of 2.7 tuberculin units/micrograms. An intradermal dose of 0.50 microgram was found to give typical delayed skin test reactions in all of 15 tuberculin-positive subjects, with a mean reaction size of 15.17 mm. Some preparations of antigen 5 were found to be unstable during lyophilization and storage; solutions of antigen 5 in diluent containing Tween 80 were stable. Mycobacterium tuberculosis antigen 5 is a potent tuberculin skin test antigen deserving of further clinical evaluation.
Antigens, Bacterial↗
The immune spectrum in patients with pulmonary tuberculosis.
The immune response to mycobacterial antigens of 65 patients with pulmonary tuberculosis was studied using delayed skin test reactions and enzyme-linked immunosorbent antibody assays. Evidence for a spectrum of immune response was found. Six of 35 patients studied during the first month of therapy had tuberculin skin test anergy. Anergy was not related to state of nutrition or extent of disease, but it may have been associated with radiographically acute disease. The 6 anergic patients had somewhat higher antibody titers to mycobacterial protein, but not polysaccharide antigens, than did nonanergic patients.
Acute Disease↗
Suppression of lymphocyte responses by tuberculous plasma and mycobacterial arabinogalactan. Monocyte dependence and indomethacin reversibility.
During tuberculosis, exposure of monocytes to circulating factors may induce the suppressor activity observed in some anergic patients. To explore this possibility, we examined the effects of plasma pooled from 28 untreated tuberculosis (TB) patients and the mycobacterial cell wall polysaccharide D-arabino-D-galactan (AG) on the in vitro function of peripheral blood mononuclear cells (PBMC) from healthy donors. In the [3H] thymidine incorporation assay, stimulated responses of PBMC incubated in culture medium supplemented with TB plasma or co-cultured with 3.0 microgram/ml AG were depressed significantly when compared with control responses. Cytotoxicity and altered kinetics of stimulated DNA synthesis did not contribute to the observed suppression. TB plasma and AG-induced suppression of the PBMC response to purified protein derivative was monocyte dependent and indomethacin reversible. In addition, TB plasma and AG directly inhibited the phytohemagglutinin-stimulated responses of T lymphocytes. In a quantitative assay of monocyte attachment to plastic, both TB plasma and AG significantly increased monocyte adherence from basal levels. These effects on monocyte adherence were reversed with indomethacin or antibody to mycobacterial polysaccharide. In addition, TB plasma passed over an immunoabsorbent column of Sepharose-linked antibody to mycobacterial polysaccharide was depleted of the suppressive and monocyte-adherence augmenting factors. 3.0 microgram/ml AG stimulated a fivefold increase in prostaglandin E2 production by cultured mononuclear cells. Our data suggest that AG circulating alone or bound in immune complexes may account for the observed effects of TB plasma. Similar in vivo exposure may contribute to the cell-mediated suppression of lymphocyte responses in tuberculosis.
Cell Adhesion↗
Recognizing and managing acute lower extremity thrombosis.
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Adult↗
Identification and preliminary purification of a protein antigen apparently specific to mycobacterium kansasii.
A highly anodal protein antigen of molecular weight approximately 10,000 daltons has been identified in culture filtrates of Mycobacterium kansasii and not in culture filtrates of 9 other mycobacterial species. Using immunoabsorbent affinity chromatography, this antigen has been purified and used to skin test guinea pigs sensitized with Mycobacterium kansasii and Mycobacterium tuberculosis. In these animals it was found to be tuberculin active and to have partial species specificity.
Animals↗
In vitro and in vivo reactivity and specificity of fractions from sonicates of Mycobacterium tuberculosis separated by gradient acrylamide gel electrophoresis.
Fractions of cell sonicates from the H37Rv strain of Mycobacterium tuberculosis prepared by gradient acrylamide gel electrophoresis (GAGE) were examined by immunoelectrophoretic analysis and tested for biologic reactivity and antigenic specificity. Each was tested for the ability to elicit delayed type hypersensitivity reactions in the skin of guinea pigs sensitized with M. tuberculosis, M. kansasii, M. intracellulare (Battey), M. scrofulaceum (Gause), and M. fortuitum. Peripheral lymphocytes from persons infected with mycobacteria were also exposed to fractions, and the increase in [3H]thymidine incorporation into DNA was used as an index of stimulation (lymphocyte transformation). Only 4 fractions (6, 11, 12, and 16) were found to be significantly more specific than the unfractionated sonicate, and none was monospecific for M. tuberculosis. The results of fused rocket immunoelectrophoresis suggested that incomplete separation of antigens and/or shared group-specific multideterminant antigens may have accounted for the observed cross-reactions.
Animals↗