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Biomedical subjects

G Kronvall

Publications and source records attributed to G Kronvall.

At least 127 records · Page 7Linked to original sources

Comparative studies of antigen 21 in Mycobacterium and Nocardia species: possible taxonomic relationships with Mycobacterium leprae.

Studies of Mycobacterium leprae, Mycobacterium tuberculosis and Nocardia caviae in comparison with each other and with other Mycobacterium and Nocardia species were performed on the basis of antigen 21 intramolecular heterogeneity. Three different antisera were used: rabbit anti-Mycobacterium smegmatis antiserum, rabbit anti-Nocardia asteroides antiserum, and a lepromatous serum pool. With reference to each of the three antiserum sources used the strains were ranked in an order of relatedness or sharing of determinants. The three antisera showed distinctly different antigen 21 antibody specificities reflecting the species origin of the immunogen. The present investigations confirmed that antigen 21 of N. caviae shares determinants with antigens from Mycobacterium strains which were not present in corresponding antigens of all other Nocardia strains tested. M. tuberculosis, as judged by antigen 21 analysis, occupies a position separate from both the slow-growing and the fast-growing mycobacterial clusters in accordance with accepted taxonomic relationships. An interesting possibility of establishing a position for M. leprae in relation to other mycobacterial species was apparent. The order of relatedness among the strains studied went from M. leprae to M. tuberculosis to N. caviae to Mycobacterium avium to Mycobacterium fortuitum, the last two being representatives of the slow-growing and fast-growing mycobacteria. It can therefore be concluded that evidence from antigen 21 analysis indicates that M. leprae is more closely related to M. tuberculosis than to the other strains investigated.

Antigens, Bacterial↗

Detection of antigen in Nocardia caviae cross-reacting with mycobacterial antigen No. 21 in M. leprae using a lepromatous leprosy serum pool as antibody reagent.

Eighteen strains of nocardia, representing five different species, were studied in crossed immunoelectrophoresis against a lepromatous leprosy serum pool for the presence of antigen No. 21. This mycobacterial antigen shows antigenic heterogeneity with species specific antigenic determinants defined in Mycobacterium leprae. All four strains of N. caviae were found to share antigen No. 21 determinants with mucobacteria. All other strains of nocardia were negative in these direct immunoprecipitation tests. When compared with M. leprae antigen No. 21, the N. caviae antigen gave a reaction of partial identify in the same way as all strains of other mycobacteria tested previously, ii.e., with spurring by the M. leprae antigen. There was a reaction of complete identity between N. caviae and M. avium-intracellulare and M. smegmatis, respectively, with the lepromatous leprosy serum pool used as the antibody source. The results suggest that N. caviae antigen No. 21 is more closely related to the corresponding antigen of the genus Mycobacterium than to the antigen No. 21 equivalent of other nocardia species tested.

Antigens, Bacterial↗

Cross-reactions between mycobacteria. II. Crossed immunoelectrophoretic analysis of soluble antigens of BCG and comparison with other mycobacteria.

Cross-reactions between Mycobacterium bovis BCG and various other mycobacteria, Nocardia asteroides, Corynebacterium pyogenes and Listeria monocytogenes were studied by incorporating antibodies against these bacteria in the intermediate gel of a crossed immunoelectrophoretic system with BCG antigen and anti-BCG antibodies. In the BCG reference system forty-four distinct antigenic components were recorded, of which thiryt-three cross-reacted with Mycobacterium tuberculosis, twenty-five with M. avium, twenty-one with M. suvalii, eighteen with M. smegmatis, Fifteen with M. nonchromogenicum, twelve with M. phlei, eight with N. anteroides and two with C. pyogenes, whereas no cross-reaction was detected with L. monocytogenes. The value of the method for characterization of mycobacterial antigens is discussed. A taxonomic system based on this method appears particularly valuable for studies of non-cultivativable mycobacteria such as M. leprae. A majority of twenty-one patients with lepromatous leprosy had anti-BCG antibodies of restricted specificity, affecting only four or five BCG antigens, although one patient had twelve anti-BCG specificities. Most of these antibodies reacted with those BCG antigens that cross-react extensively with other mycobacteria.

Antibodies, Bacterial↗

Immunoglobulin-binding structure on bovine group G streptococci different from type III Fc receptors on human group G streptococci.

The immunoglobulin G (IgG)-binding capacity of 54 group G streptococci of human and bovine origin was investigated. Of 20 human strains, 17 carried a surface component which could combine with human IgG and bovine IgG1 and IgG2. Inhibition experiments with unlabeled human IgG and with a panel of animal sera revealed that the same surface component was involved in the binding of human as well as bovine immunoglobulins. Of 16 beta-hemolytic bovine group G streptococci, 13 reacted with human IgG but not with bovine IgG1 or IgG2. This binding structure was different from the type III Fc reactivity found in human group G streptococci. All human strains, including the three IgG Fc-nonreactive strains, fermented trehalose, in contrast to all bovine beta-hemolytic strains, which were negative. Immunoglobulin Fc reactivity is thus a feature not only of human strains but also of some bovine strains.

Animals↗

Specific absorption of human serum albumin, immunoglobulin A, and immunoglobulin G with selected strains of group A and G streptococci.

Five gram-positive bacterial strains were selected for absorption studies of human serum samples. Strain AR1 (group A, M-type 1) and G148 (group G), with strong immunoglobulin G (IgG) binding capacities, and strain AW43 (group A, M-type 60), binding both IgA1 and IgA2, were compared with Staphylococcus aureus Cowan I and with Staphylococcus epidermidis L603. Both AR1 and G148 were capable of completely absorbing out serum IgG. In contrast, S. aureus Cowan I left a fraction unabsorbed, as expected from its known lack of IgG3 binding. Strain AW43 absorbed out all serum IgA, using a 10-microliter bacterial pellet for 20 microliter of serum. Serum IgM levels were slightly reduced by S. aureus Cowan I absorption. On the basis of the experiments, a bacterial mixture was designed consisting of S. aureus Cowan I and group A streptococcus strains AR1 and AW43, with absorption characteristics suitable for use in discriminating between early IgM and late IgG and IgA immune responses in routine serological work. A new type of bacteria-mammalian protein binding was discovered. Human serum albumin was completely absorbed out by strain G148 and to a lesser extent by strain AR1 and AW43. S. aureus Cowan I and S. epidermidis were negative. The binding capacity of G148 for albumin equalled that of Cowan I for IgG. The binding pattern of albumin to the strains was different from those of IgG, IgA, IgM, fibrinogen, haptoglobin, or aggregated beta 2-microglobulin and therefore seems to represent another type of bacterial-mammalian interaction with a specific albumin receptor on the surface of streptococci.

Absorption↗

Interactions between human serum proteins and oral streptococci reveal occurrence of receptors for aggregated beta 2-microglobulin.

A total of 31 strains of oral streptococci representing Streptococcus mutans, Streptococcus sanguis, Streptococcus mitior, Streptococcus salivarius, and Streptococcus milleri were tested for possible binding of human immunoglobulins G, G1, G2, G3, G4, A1, A2, M1, and M2 and haptoglobin, hemoglobin, fibrinogen, and aggregated beta 2-microglobulin. Radiolabeled beta 2-microglobulin in aggregated form showed affinity for 20 of the 31 strains tested. Binding activity for the protein was found in strains belonging to all five species. The bacterial receptor was resistant to trypsin. Monomeric, unlabeled beta 2-microglobulin did not interfere with the binding of the aggregated form. Of the other proteins tested, only the immunoglobulin A1 protein showed positive binding, and that was only with a single strain of S. milleri. beta 2-Microglobulin is present on all nucleated cell membranes in vivo. The reaction between aggregated beta 2-microglobulin and oral streptococci is a new type of human-bacterium interaction which should be considered in studies of bacterial adherence.

Beta-Globulins↗

Lymphocyte transformation test in healthy contacts of patients with leprosy. I. Influence of exposure to leprosy within a household.

Fifty-three household contacts of lepromatous patients, 37 household contacts of tuberculoid patients, and 91 control persons were examined with the lymphocyte transformation test (LTT) for their responses to whole and sonicated antigen preparations from M. leprae, to BCG, M. avium, M. gordonae, and phytohemagglutinin (PHA). The study was carried out in the Gurage area of Ethiopia in 15 households with a leprosy patient and 15 matched control households. Household contacts of lepromatous patients showed significantly greater LTT responses to antigens from M. leprae than the controls, whereas household contacts of tuberculoid patients did not respond differently from controls. Household contacts of lepromatous patients had significantly greater responses to M. leprae antigens when the index patients were "active," i.e., highly bacilliferous, than when they were "inactive," i.e., having a low bacillary load. The degree of sensitization, as indicated by the LTT response, in different exposure groups paralleled the degree of probable infectivity of the index patient. A preparation of antigen from whole M. leprae proved to be more sensitive and more specific in the LTT than did a sonicated preparation. A significant degree of cross-reactivity was found among the various mycobacteria in their LTT responses.

Adolescent↗

Lymphocyte transformation test in healthy contacts of patients with leprosy. II. Influence of consanguinity with the patient, sex, and age.

The study was carried out in the Gurage area of Ethiopia, where 53 household contacts of lepromatous patients, 37 household contacts of tuberculoid patients, and 91 control persons were examined with the lymphocyte transformation test (LTT) for their responses to whole and sonicated antigen preparation from M. leprae to BCG, M. avium, M. gordonae and phytohemagglutinin. The potential influence of host factors, namely the state of consanguinity with the leprosy patient, sex and age on the LTT responses was evaluated. In the 35 household contacts of "active," i.e., highly bacilliferous, lepromatous patients, consanguinity with a lepromatous patient was not associated with a significant depression of the LTT responses to M. leprae antigens. Male household contacts of active lepromatous patients showed significantly greater LTT responses to M. leprae antigens than female household contacts. Possible confounding factors for this finding are discussed. Sensitization of M. leprae antigens was present already in a high proportion of the 6 to 14 year old household contacts of active lepromatous patients, which was the youngest age group examined in our study. No significant results were found in any of the other patient contact groups with regard to the host factors examined.

Adolescent↗

Fibrinogen binding structures in beta-hemolytic streptococci group A, C, and G. Comparisons with receptors for IgG and aggregated beta 2-microglobulin.

Binding of radiolabelled fibrinogen was measured to 197 strains of 16 different bacterial species. All streptococcal strains belonging to groups A, C, and G isolated from human sources were strongly positive. S. aureus strains showed low binding values. Occasional group B streptococci were positive. Reactive strains were also noted among group C streptococci of animal origin, Streptococcus zooepidemicus and Str. equii, and bovine beta-hemolytic group G streptococci. Bovine alpha-hemolytic group G strains as well as the remaining seven species of human origin were all negative. Inhibition experiments and correlation studies indicated that the streptococcal receptor for fibrinogen was different from immunoglobulin Fc binding reactivity. Comparisons with the newly discovered beta 2-microglobulin binding factor showed that trypsin concentrations which destroyed this receptor left the fibrinogen receptor intact. Although the two receptors correlate in strain population studies and show competition for binding the difference in trypsin sensitivity indicates that they represent two different structural entities. Both receptors might serve as basic markers for M-protein like surface components of Gram positive cocci.

Beta-Globulins↗

Mycobacterium leprae specific antibodies detected by radioimmunoassay.

A radioimmunoassay was developed for demonstration of antibodies against M. leprae specific antigenic determinants. The specificity of the assay was tested with hyperimmune rabbit antisera against other mycobacteria and shown to be very high. The titre of M. leprae specific antibodies in a lepromatous serum pool was 10(5). Sixty-one of sixty-two lepromatous sera, all of twelve borderline sera and twenty of forty-eight tuberculoid sera were positive in the assay, whereas all of thirty-eight control sera from tuberculin positive individuals from a leprosy non-endemic area were negative. Aplication as a diagnostic test for subclinical infection with M. leprae is discussed. The principle of the test appears promising for serological distinction between pulmonary infection with M. tuberculosis and other mycobacteria.

Antibodies, Bacterial↗

Binding of aggregated human beta2-microglobulin to surface protein structure in group A, C, and G streptococci.

A novel mammalian-microbial "short circuit" has been demonstrated between aggregated human beta2-microglobulin and group A, C, and G streptococci. Bacteria belonging to nine gram-positive and three gram-negative species were tested for binding of radiolabeled beta2-microglobulin. All 10 individual strains of group A streptococci showed a high degree of reactivity with aggregated human beta2-microglobulin. Among 27 group C and 28 group G streptococci, 9 and 6 strains, respectively, were highly reactive, whereas the remaining strains showed a lower, but definite level of beta2-microglobulin binding. Of 11 group B streptococci, 4 were slightly positive. All strains among the other eight species were completely negative. Simultaneous testing of A, C, and G streptococci for immunoglobulin binding showed a lack of correlation between type II and III Fc reactivity and beta2-microglobulin binding. There was no inhibition of uptake of aggregated beta2-microglobulin to reactive strains when excess amounts of human immunoglobulin were added. The beta2-microglobulin-binding surface structure was found to be markedly sensitive to trypsin digestion. The relative trypsin resistance of the immunoglobulin-binding protein in the digestion experiments further demonstrated the dissociation between these two reactivities.

Bacterial Proteins↗

Non-equilibrium enzyme immunoassay of gentamicin.

We describe a fast, accurate non-equilibrium enzyme immunoassay for serum gentamicin. With use of isokinetic conditions an assay takes a total of 12 min, including regeneration of the immunosorbent. The coefficient of correlation between the present method and conventional microbiological assays was 0.98.

Catalase↗

Discrepancy between Clq deviation and Raji cell tests in detection of circulating immune complexes in patients with leprosy.

Samples of serum from 45 patients with different clinical forms of leprosy and from 17 patients with systemic lupus erythematosus were studied in parallel for circulating immune complexes with use of two different in vitro tests adjusted to the same degree of sensitivity. The Clq deviation test relied upon the reaction of the complement component Clq with immune complexes. The Raji cell test detected complement-fixed immune complexes that bound to the complement receptors on cultured, bone marrow-derived lymphocyte-like Raji cells. Thirty (67%) of 45 patients with leprosy showed immune complexes according to the Clq deviation test; however, only two (7%) of the 30 samples of sera with positive Clq test results were positive by the Raji cell test. In contrast, 54% of 13 samples of sera from patients with systemic lupus erythematosus positive by the Clq test were positive according to the Raji cell test. Since Clq is known to react with DNA as well as with bacterial antigens, the Clq reaction may in fact be detecting antigenemia in many instances. Considerable caution is warranted in application of sensitive screening tests for assay of circulating immune complexes in various states of infectious diseases.

Antibodies↗

Differential staining of bacteria in clinical specimens using acridine orange buffered at low pH.

Optimal conditions for acridine orange staining of air dried and methanol fixed bacteria on glass slides were studied. The pH of the staining buffer did not influence the fluorescence of an S. aureus and an E. coli strain at dye concentrations of 25-50 mg per litre. 81 bacterial strains representing 15 different species were stained with acridine orange under standard conditions, all strains showing orange fluorescence. The pH of the buffer influenced markedly the staining patterns of human cells and tissue materials, as represented by smears of peripheral blood, buccal scrapings, urethral secretions and tracheal exudates. The fluorescence obtained ranged from low intensity green at low pH values to bright orange at neutral and alkaline pH. This variability indicated a possibility of designing conditions for a differential staining method for the detection of bacteria in clinical specimens. The differential staining effect with a low pH in the buffer was confirmed on smears of buccal scrapings, cerebrospinal fluid samples and urethral secretions, showing orange fluorescence of the bacteria present and green-to-yellow fluorescence of background material, cells and tissue debris.

Acridines↗

Common antigen of Mycobacterium leprae, M. lepraemurium, M. avium, and M. fortuitum in comparative studies using two different types of antisera.

No. 21 mycobacterial antigens of Mycobacterium lepraemurium, M. avium, M. fortuitum, and M. leprae were compared in crossed immunoelectrophoresis using two different antibody sources, a serum pool from lepromatous leprosy patients (LSII) and a rabbit anti-M. smegmatis antiserum. M. lepraemurium, like M. avium, was found to contain the 21 A and 21 C determinants. M. fortuitum contained in addition a new type of determinant, 21 D.M. leprae antigen no. 21 carried the A as well as the B dertminants, the latter found so far only in the leprosy bacillus. The separate taxonomic position of M. leprae, suggested by earlier studies of the no. 21 antigen, is further supported by the present results, which also demonstrate the potential use of submolecular heterogeneity for such investigations.

Animals↗

Heterogeneity of nonimmune immunoglobulin Fc reactivity among gram-positive cocci: description of three major types of receptors for human immunoglobulin G.

Two hundred and thirty strains of various gram-positive cocci were tested for quantitative, nonimmune binding of radiolabeled human polyclonal immunoglobulin G (IgG). The majority of coagulase-positive staphylococci and streptococci belonging to serogroups C and G showed a high uptake of IgG. The binding of immunoglobulin to group A streptococci was considerably less, with a number of strains completely negative. None of the pneumococcal or the group B or D streptococcal strains displayed any binding capacity. Heterogeneity of the IgG reactivity of various reactive strains was studied in an inhibition assay using 10 different animal serum pools. Three different inhibition patterns were seen, each of them revealing a striking degree of homogeneity within single bacterial species. Staphylococcus aureus and group A streptococci, respectively, constituted two homogeneous groups which differed markedly from each other and from C and G streptococci. No differences were observed between group C and G streptococci. Based on the profound differences between these homogeneous groups, three major types of Fc receptors could be defined. Type I and II Fc receptors were found on S. aureus and on group A streptococci, respectively. Fc receptor type III represented the immunoglobulin-binding structure of both group C and G streptococci.

Animals↗

Antibody response in rabbits to immunization with Mycobacterium leprae.

Mycobacterium leprae purified from liver tissue of an infected armadillo (the A/10 preparation) was tested for antigenic composition by immunization of rabbits and characterization of the antibody response by crossed immunoelectrophoresis. The rabbit antisera detected seven distinct components in the M. leprae preparation. This number is far lower than in similar experiments with other mycobacteria. The M. leprae sonic extract gave far fewer lines after polyacrylamide gel electrophoresis and staining with Coomassie brillant blue than sonic extracts prepared from BCG, M. smegmatis, and M. phlei adjusted to the same protein concentration based on the Folin assay. The seven components detected in M. leprae cross-reacted extensively with M. avium, BCG, M. lepraemurium, M. smegmatis, and Nocardia asteroides. The seven components are involved in immune reactions in leprosy; antibodies against all of them were demonstrated in sera from patients with lepromatous leprosy, but the specificity of the antibodies varied from patient to patient. The reason for the demonstration of so few antigenic components and some of the implications of these findings for the use of armadillo-grown M. leprae to develop specific skin test reagents and in other aspects of leprosy research are discussed.

Animals↗