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. 1993 Jan;61(1):8–12. doi: 10.1128/iai.61.1.8-12.1993

Effect of pH and osmolality on in vitro phagocytosis and killing by neutrophils in urine.

R A Gargan 1, J M Hamilton-Miller 1, W Brumfitt 1
PMCID: PMC302681  PMID: 8418067

Abstract

Phagocytosis and intracellular killing of two strains of Escherichia coli and a Staphylococcus saprophyticus by polymorphonuclear neutrophils (PMN) in pooled sterile urine at three osmolalities (800, 485, and 200 mosM/kg of H2O) between pHs 5 and 8 was investigated. Urine at 800 mosM virtually abolished phagocytosis of both E. coli strains, regardless of pH, and reduced the phagocytosis of S. saprophyticus to 30%; no killing of any organisms took place at this osmolality. On the other hand, phagocytosis was a good in urine as in Hanks balanced salt solution at both 485 and 200 mosM between pHs 6 and 8. Phagocytosis of all three strains was virtually abolished at pH 5. Killing of the strains by PMN was optimal between pHs 6.5 and 7.5 in urine at 485 mosM (being at least 90% of the control values in Hanks balanced salt solution), whereas at 200 mosM killing was reduced to 50 to 70% of these values. Reduced killing of all three strains occurred at pH 8, whereas at pH 6 only S. saprophyticus was killed. Thus, the bactericidal activity of PMN in urine was more sensitive than phagocytic function to alterations in pH. The dominant modulating factor affecting PMN function in urine of 500 mosM or less was pH, but osmolality had a greater influence at 800 mosM. Thus, raising the pH of urine and reducing the osmolality may increase the ability of natural defense mechanisms to eliminate infecting organisms.

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Selected References

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  1. Akerlund A. S., Ahlstedt S., Hanson L. A., Jodal U. Antibody responses in urine and serum against Escherichia coli O antigen in childhood urinary tract infection. Acta Pathol Microbiol Scand C. 1979 Feb;87C(1):29–36. [PubMed] [Google Scholar]
  2. Asscher A. W., Sussman M., Waters W. E., Davis R. H., Chick S. Urine as a medium for bacterial growth. Lancet. 1966 Nov 12;2(7472):1037–1041. doi: 10.1016/s0140-6736(66)92023-x. [DOI] [PubMed] [Google Scholar]
  3. Bryant R. E., Sutcliffe M. C., McGee Z. A. Human polymorphonuclear leukocyte function in urine. Yale J Biol Med. 1973 Apr;46(2):113–124. [PMC free article] [PubMed] [Google Scholar]
  4. Burdof D. W. Quantitative studies on urinary immunoglobulins in hospital patients, including patients with urinary tract infection. Clin Exp Immunol. 1970 Feb;6(2):189–196. [PMC free article] [PubMed] [Google Scholar]
  5. CHERNEW I., BRAUDE A. I. Depression of phagocytosis by solutes in concentrations found in the kidney and urine. J Clin Invest. 1962 Oct;41:1945–1953. doi: 10.1172/JCI104652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. COX C. E., HINMAN F., Jr Experiments with induced bacteriuria, vesical emptying and bacterial growth on the mechanism of bladder defense to infection. J Urol. 1961 Dec;86:739–748. doi: 10.1016/S0022-5347(17)65257-1. [DOI] [PubMed] [Google Scholar]
  7. Cattell W. R., Fry I. K., Spiro F. I., Sardeson J. M., Sutcliffe M. B., O'Grady F. Effect of diuresis and frequent micturition on the bacterial content of infected urine: a measure of competence of intrinsic hydrokinetic clearance mechanisms. Br J Urol. 1970 Jun;42(3):290–295. doi: 10.1111/j.1464-410x.1970.tb11922.x. [DOI] [PubMed] [Google Scholar]
  8. Dooley D. C., Takahashi T. The effect of osmotic stress on the function of the human granulocyte. Exp Hematol. 1981 Aug;9(7):731–741. [PubMed] [Google Scholar]
  9. 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 Jul 6;121(1):105–113. doi: 10.1016/0022-1759(89)90425-0. [DOI] [PubMed] [Google Scholar]
  10. Fukushi Y., Orikasa S. The role of intravesical polymorphonuclear leukocytes in experimental cystitis. Invest Urol. 1981 May;18(8):471–474. [PubMed] [Google Scholar]
  11. Gabig T. G., Bearman S. I., Babior B. M. Effects of oxygen tension and pH on the respiratory burst of human neutrophils. Blood. 1979 Jun;53(6):1133–1139. [PubMed] [Google Scholar]
  12. Gargan R. A., Brumfitt W., Hamilton-Miller J. M. Failure of water to lyse polymorphonuclear neutrophils completely. Role of pH and implications for assessment of bacterial killing. J Immunol Methods. 1989 Nov 30;124(2):289–291. doi: 10.1016/0022-1759(89)90368-2. [DOI] [PubMed] [Google Scholar]
  13. Gargan R. A., Brumfitt W., Hamilton-Miller J. M. Pre-opsonisation of Escherichia coli induces resistance to neutrophil killing in serum and urine: relationship to growth phase. J Med Microbiol. 1991 Jul;35(1):12–17. doi: 10.1099/00222615-35-1-12. [DOI] [PubMed] [Google Scholar]
  14. Gillon G., Small M., Medalia O., Aronson M. Sequential study of bacterial clearance in experimental cystitis. J Med Microbiol. 1984 Dec;18(3):319–326. doi: 10.1099/00222615-18-3-319. [DOI] [PubMed] [Google Scholar]
  15. Horwitz M. A., Silverstein S. C. Influence of the Escherichia coli capsule on complement fixation and on phagocytosis and killing by human phagocytes. J Clin Invest. 1980 Jan;65(1):82–94. doi: 10.1172/JCI109663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kaye D. Antibacterial activity of human urine. J Clin Invest. 1968 Oct;47(10):2374–2390. doi: 10.1172/JCI105921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lancaster M. G., Allison F., Jr Studies on the pathogenesis of acute inflammation. VII. The influence of osmolality upon the phagocytic and clumping activity by human leukocytes. Am J Pathol. 1966 Dec;49(6):1185–1200. [PMC free article] [PubMed] [Google Scholar]
  18. Maeda S., Deguchi T., Kanimoto Y., Kuriyama M., Kawada Y., Nishiura T. Studies on the phagocytic function of urinary leukocytes. J Urol. 1983 Feb;129(2):427–429. doi: 10.1016/s0022-5347(17)52131-x. [DOI] [PubMed] [Google Scholar]
  19. Matsumoto T., Kumazawa J., van der Auwera P. Suppression of leukocyte function and intracellular content of ATP in hyperosmotic condition comparable to the renal medulla. J Urol. 1989 Aug;142(2 Pt 1):399–402. doi: 10.1016/s0022-5347(17)38772-4. [DOI] [PubMed] [Google Scholar]
  20. Matsumoto T., van der Auwera P., Watanabe Y., Tanaka M., Ogata N., Naito S., Kumazawa J. Neutrophil function in hyperosmotic NaCl is preserved by phosphoenol pyruvate. Urol Res. 1991;19(4):223–227. doi: 10.1007/BF00305299. [DOI] [PubMed] [Google Scholar]
  21. Suzuki Y., Fukushi Y., Orikasa S., Kumagai K. Opsonic effect of normal and infected human urine on phagocytosis of Escherichia coli and yeasts by neutrophils. J Urol. 1982 Feb;127(2):356–360. doi: 10.1016/s0022-5347(17)53781-7. [DOI] [PubMed] [Google Scholar]
  22. 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]. Nihon Hinyokika Gakkai Zasshi. 1986 Apr;77(4):574–580. doi: 10.5980/jpnjurol1928.77.4_574. [DOI] [PubMed] [Google Scholar]
  23. Thomas V. L., Forland M., Shelokov A. Immunoglobulin levels and antibody-coated bacteria in urines from patients with urinary tract infections. Proc Soc Exp Biol Med. 1975 Apr;148(4):1198–1201. doi: 10.3181/00379727-148-38716. [DOI] [PubMed] [Google Scholar]

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