1887

Abstract

Summary

Experiments were performed to determine the effects of products of bacterial growth (including endotoxin) on phagocytosis and intracellular killing by polymorphonuclear leucocytes (PMNL) in urine. Bacteriologically filtered supernates of two strains of grown in urine were added in varying amounts to mixtures of PMNL and , also in urine. Phagocytosis of the two strains was reduced from > 90% in controls to 66% and 48%, respectively, in the presence of undiluted culture filtrate (containing endotoxin 2—2.5 μg/ml). Intracellular killing was also decreased and was abolished by dilutions corresponding to endotoxin concentrations of 0.6 and 0.75 μg/ml. When PMNL exposed to these inhibitory dilutions were resuspended in fresh urine, their phagocytic ability was fully restored and 13—24% of their killing activity was regained. A minimum concentration of commercially purified endotoxin of 200 μg/ml was required to abolished PMNL killing, with phagocytosis uninhibited. The results strongly suggest that bacterial growth metabolites, not endotoxin, are responsible for the depression of phagocytosis and intracellular killing in infected urine. A moderate dilution of the bacterial products in urine permits good PMNL function. Extrapolating this to the clinical situation, diluting the urine by water loading (as recommended for patients with urinary infections) should ensure efficient activity of PMNL under in-vivo conditions providing urinary pH and osmolality are not adversely affected.

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1993-07-01
2022-05-26
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References

  1. Chemew I, Braude AI. Depression of phagocytosis by solutes in concentrations found in the kidney and urine. J Clin Invest 1962; 41:1945–1953
    [Google Scholar]
  2. Suzuki Y, Toyota S, Magima K, Fukushi Y, Orikasa S. Studies of anti-bacterial defense mechanism of urinary bladder. 1) Activity of phagocytosis of PMN in urine. 2) Viability, cell population and activity of phagocytosis of urinary PMN. Nippon Hinyokika Gakkai Zasshi 1986; 77:574–580 (English abstract)
    [Google Scholar]
  3. Bryant RE, Sutcliffe MC, McGee ZA. Human poly morphonuclear leukocyte function in urine. Yale J Biol Med 1973; 46:113–124
    [Google Scholar]
  4. Gargan RA, Hamilton-Miller JMT, Brumfitt W. Effect of pH and osmolality on in vitro phagocytosis and killing by neutrophils in urine. Infect Immun 1993; 61:8–12
    [Google Scholar]
  5. Wilson ME. Effects of bacterial endotoxins on neutrophil function. Rev Infect Dis 1985; 7:404–418
    [Google Scholar]
  6. Aida Y, Pabst MJ. Neutrophil responses to lipopolysaccharide. Effect of adherence on triggering and priming of the respiratory burst. J Immunol 1991; 146:1271–1276
    [Google Scholar]
  7. Guthrie LA, McPhail LC, Henson PM, Johnston RB. Priming of neutrophils for enhanced release of oxygen metabolites by bacterial lipopolysaccharide. Evidence for increased activity of the superoxide-producing enzyme. J Exp Med 1984; 160:1656–1671
    [Google Scholar]
  8. Cohn ZA, Morse SI. Functional and metabolic properties of polymorphonuclear leucocytes II. The influence of a lipopolysaccharide endotoxin. J Exp Med 1960; 111:689–704
    [Google Scholar]
  9. Henricks PAJ, Van Der Tol ME, Thyssen RMWM, Van Asbeck S, Verhoef J. Escherichia coli lipopolysaccharides diminish and enhance cell function of human polymorphonuclear leukocytes. Infect Immun 1983; 41:294–301
    [Google Scholar]
  10. Proctor RA. Endotoxin in vitro interactions with human neutrophils: depression of chemiluminescence, oxygen consumption, superoxide production, and killing. Infect Immun 1979; 25:912–921
    [Google Scholar]
  11. Maeda S, Deguchi T, Kanimoto Y, Kuriyama M, Kawada Y, Nishiura T. Studies on the phagocytic function of urinary leukocytes. J Urol 1983; 129:427–429
    [Google Scholar]
  12. Jorgensen JH, Jones PM. Comparative evaluation of the Limulus assay and the direct gram stain for detection of significant bacteriuria. Am J Clin Pathol 1975; 63:142–148
    [Google Scholar]
  13. Eggleton P, Gargan R, Fisher D. Rapid method for the isolation of neutrophils in high yield without the use of dextran or density gradient polymers. J Immunol Methods 1989; 121:105–113
    [Google Scholar]
  14. Ford WL. The preparation and labelling of lymphocytes. In: Weir DM. (ed) Handbook of experimental immunology, vol 2 Oxford: Blackwell Scientific Publications; 1978231–23
    [Google Scholar]
  15. Gargan RA, Brumfitt W, Hamilton-Miller JMT. Pre-opsonisation of Escherichia coli induces resistance to neutrophil killing in serum and urine: relationship to growth phase. J Med Microbiol 1991; 35:12–17
    [Google Scholar]
  16. Gargan RA, Brumfitt W, Hamilton-Miller JMT. Failure of water to lyse polymorphonuclear neutrophils completely: role of pH and implications for assessment of bacterial killing. J Immunol Methods 1989; 124:289–291
    [Google Scholar]
  17. Horwitz MA, Silverstein SC. Influence of the Escherichia coli capsule on complement fixation and on phagocytosis and killing by human phagocytes. J Clin Invest 1980; 65:82–94
    [Google Scholar]
  18. Creamer HR, Hunter N, Bullock WW, Gabler WL. Con current lipopolysaccharide enhances chemotactic response of human polymorphonuclear leukocytes to bacterial chemotaxin. Inflammation 1991; 15:201–211
    [Google Scholar]
  19. Bishop MB, Baltch AL, Hill LA, Smith RP, Lutz F, Pollack M. The effect of Pseudomonas aeruginosa cytotoxin and toxin A on human polymorphonuclear leukocytes. J Med Microbiol 1987; 24:315–324
    [Google Scholar]
  20. Rotstein OD, Wells CL, Pruett TL, Sorenson JJ, Simmons RL. Succinic acid production by Bacteroides fragilis. A potential bacterial virulence factor. Arch Surg 1987; 122:93–98
    [Google Scholar]
  21. Bortolussi R, Vandenbroucke-Grauls CMJE, Van Asbeck BS, Verhoef J. Relationship of bacterial growth phase to killing of Listeria monocytogenes by oxidative agents generated by neutrophils and enzyme systems. Infect Immun 1987; 55:3197–3203
    [Google Scholar]
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