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    Neutrophil function studies with respect to antibiotic tolerant staphylococcus aureus and patients with recurring infection

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    Authors
    Raynor, Robert H
    Issue Date
    1980-08
    URI

    http://hdl.handle.net/10675.2/622506
    
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    Abstract
    Polymorphonuclear neutrophils (PMNs) from s~ven patients who experien<;: ed recurring staphylococcal infections, were tested in order to establish if measurements of phagocytic and bactericidal capacity would reveal a functional cause for the repeated episodes of disease. Using a microprocedure developed for this 'purpose, the results for six of these patients were found to be comparable to normal values. PMNs from one patient, however, showed a deficiency in their ability to both phagocytize and kill s. aureus. This defect could not be attributed to serum deficiencies or ascribed to any previously defined class of nel.ltrophil function. In other studies, six clinical isolates of Staphylococcus aureus were compared for their relative susceptibilities to the killing effects of oxacillin. Three of the strains had minimum bactericidal concentrations which were >10 times the minimum bacteriostatic concentration for this antibiotic and were designated tolerant (Tol+). The other st;r-ains had minimum bactericidal concentrations which were comparable to the minimum bacteriostatic concentration (Tol-). Lysis curves of these strains revealed that the Tal+ strains exhibited a diminished rate of lysis when inhibited by oxacillin. This reduced rate of lysis was reflected also in a reduced rate of viability loss when the cells were exposed to oxacillin. Cells of each phenotype, previously labeled with [ 14C]Glycerol, secreted radioactivity when inhibited by oxacillin. However,. the Tal+ strains rele.ased over twice as much labe 1 as the Talstrains. The behavior of 60 to 65%_ of the labeled material released by. inhibited cells during both sodium dodecyl sulfate gel electrophoresis and Sepharose 6B chromatography corresponded to that of lipoteichoic acid. When the major component of secreted material was added to oxacillin-inhibited Tol- strains, an inhibition of the lytic response was observed. These results suggest that oxacillin tolerance in S. aureus could be related to the enhanced secretion of an autolysin inhibitor, such as lipoteichoic acid. Several investigators have recently reported an increase in the severity of infections due to Tol+ strains. Since the enhanced'excretion of LTA by Tol+ staphylococci is the only knownphenotypJ.c difference between these two strains, the effect of this molecule on neutrophil function was measured using the microprocedure. The addition of LTA to the incubation mix·ture res1..1lted in a dose dependent inhibition of phagocytosis •. The decrease~ uptake of. S. aureus in the presence of LTA was accompanied by a corresponding increase in the number of organisms surviving in the presence of the phagocytes. Additional studies suggested that LTA interferes with the process' of opsonization, prior to engulfment.
    Affiliation
    Depatment of Cell and Molecular Biology
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