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. 1986 Mar;51(3):936–941. doi: 10.1128/iai.51.3.936-941.1986

Neutrophil activity in abscess-bearing mice: comparative studies with neutrophils isolated from peripheral blood, elicited peritoneal exudates, and abscesses.

P H Hart, L K Spencer, M F Nulsen, P J McDonald, J J Finlay-Jones
PMCID: PMC260989  PMID: 3512441

Abstract

Intraabdominal abscesses were induced in mice by intraperitoneal inoculation of Bacteroides fragilis and Escherichia coli plus bran as the abscess-potentiating agent. Six- or seven-day-old abscesses were mechanically disaggregated in buffer, and the cells obtained were fractionated on discontinuous Percoll density gradients. Neutrophil populations of different density, each approximately 90% pure, were isolated. When the abscess-derived neutrophils were subsequently incubated with normal serum in vitro under aerobic conditions, the viability of the gram-negative bacteria that had been phagocytosed within the abscess did not change significantly. This anergy to intracellular bacteria (on subsequent incubation in vitro under optimal conditions for phagocytic killing) was also found for neutrophils that had been obtained from abscesses induced by a mixture that included Proteus mirabilis plus B. fragilis and from those induced by E. coli plus P. mirabilis. While unable to significantly kill intracellular organisms that had been phagocytosed in vivo, the abscess-derived neutrophils could engulf and kill organisms to which they were exposed in vitro. Neutrophils from abscesses induced by P. mirabilis only plus bran killed that organism introduced in vitro significantly more effectively than the organisms that had been engulfed in vivo. In contrast, neutrophils from abscesses induced by the gram-positive organism Staphylococcus aureus plus bran were able to kill their intracellular organisms on subsequent incubation in vitro as effectively as they could kill added S. aureus. Neutrophils isolated from the peripheral blood and from induced peritoneal exudates of abscess-bearing mice were able to phagocytose and kill organisms in vitro with greater efficiency than abscess-derived neutrophils. The mechanism whereby neutrophils from abscesses induced by the gram-positive organism S. aureus can kill the organisms phagocytosed in vivo on subsequent in vitro incubation, in contrast to the relative anergy to their intracellular organisms displayed by neutrophils derived from abscesses induced by combinations of gram-negative bacteria, is not known.

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Selected References

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  1. Barbour A. G., Allred C. D., Solberg C. O., Hill H. R. Chemiluminescence by polymorphonuclear leukocytes from patients with active bacterial infection. J Infect Dis. 1980 Jan;141(1):14–26. doi: 10.1093/infdis/141.1.14. [DOI] [PubMed] [Google Scholar]
  2. Dooley D. C., Simpson J. F., Meryman H. T. Isolation of large numbers of fully viable human neutrophils: a preparative technique using percoll density gradient centrifugation. Exp Hematol. 1982 Aug;10(7):591–599. [PubMed] [Google Scholar]
  3. Finlay-Jones J. J., Hill N. L., Nulsen M. F., Pruul H., McDonald P. J. Opsonins in normal mouse serum for the phagocytic killing of Proteus mirabilis by murine neutrophils. Br J Exp Pathol. 1984 Dec;65(6):711–718. [PMC free article] [PubMed] [Google Scholar]
  4. Hart P. H., Spencer L. K., McDonald P. J., Finlay-Jones J. J. Evaluation of intracellular killing of bacteria by enriched populations of mouse peritoneal exudate neutrophils. Aust J Exp Biol Med Sci. 1985 Aug;63(Pt 4):361–370. doi: 10.1038/icb.1985.42. [DOI] [PubMed] [Google Scholar]
  5. Ingham H. R., Sisson P. R., Middleton R. L., Narang H. K., Codd A. A., Selkon J. B. Phagocytosis and killing of bacteria in aerobic and anaerobic conditions. J Med Microbiol. 1981 Nov;14(4):391–399. doi: 10.1099/00222615-14-4-391. [DOI] [PubMed] [Google Scholar]
  6. Jones G. R., Gemmell C. G. Impairment by Bacteroides species of opsonisation and phagocytosis of enterobacteria. J Med Microbiol. 1982 Aug;15(3):351–361. doi: 10.1099/00222615-15-3-351. [DOI] [PubMed] [Google Scholar]
  7. Leijh P. C., van den Barselaar M. T., Daha M. R., van Furth R. Participation of immunoglobulins and complement components in the intracellular killing of Staphylococcus aureus and Escherichia coli by human granulocytes. Infect Immun. 1981 Sep;33(3):714–724. doi: 10.1128/iai.33.3.714-724.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Namavar F., Verweij A. M., Bal M., van Steenbergen T. J., de Graaff J., MacLaren D. M. Effect of anaerobic bacteria on killing of Proteus mirabilis by human polymorphonuclear leukocytes. Infect Immun. 1983 Jun;40(3):930–935. doi: 10.1128/iai.40.3.930-935.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nulsen M. F., Finlay-Jones J. J., Skinner J. M., McDonald P. J. Intra-abdominal abscess formation in mice: quantitative studies on bacteria and abscess-potentiating agents. Br J Exp Pathol. 1983 Aug;64(4):345–353. [PMC free article] [PubMed] [Google Scholar]
  10. Onderdonk A. B., Bartlett J. G., Louie T., Sullivan-Seigler N., Gorbach S. L. Microbial synergy in experimental intra-abdominal abscess. Infect Immun. 1976 Jan;13(1):22–26. doi: 10.1128/iai.13.1.22-26.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pember S. O., Barnes K. C., Brandt S. J., Kinkade J. M., Jr Density heterogeneity of neutrophilic polymorphonuclear leukocytes: gradient fractionation and relationship to chemotactic stimulation. Blood. 1983 Jun;61(6):1105–1115. [PubMed] [Google Scholar]
  12. Sheridan J. W., Finlay-Jones J. J. Studies on a fractionated murine fibrosarcoma: a reproducible method for the cautious and a caution for the unwary. J Cell Physiol. 1977 Mar;90(3):535–552. doi: 10.1002/jcp.1040900316. [DOI] [PubMed] [Google Scholar]
  13. Takamori K., Yamashita T. Biochemical properties of polymorphonuclear neutrophils from venous blood and peritoneal exudates of rabbits. Infect Immun. 1980 Aug;29(2):395–400. doi: 10.1128/iai.29.2.395-400.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Tofte R. W., Peterson P. K., Schmeling D., Bracke J., Kim Y., Quie P. G. Opsonization of four Bacteroides species: role of the classical complement pathway and immunoglobulin. Infect Immun. 1980 Mar;27(3):784–792. doi: 10.1128/iai.27.3.784-792.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ulmer A. J., Flad H. D. Discontinuous density gradient separation of human mononuclear leucocytes using Percoll as gradient medium. J Immunol Methods. 1979;30(1):1–10. doi: 10.1016/0022-1759(79)90268-0. [DOI] [PubMed] [Google Scholar]
  16. Wade B. H., Kasper D. L., Mandell G. L. Interactions of Bacteroides fragilis and phagocytes: studies with whole organisms, purified capsular polysaccharide and clindamycin-treated bacteria. J Antimicrob Chemother. 1983 Oct;12 (Suppl 100):51–62. doi: 10.1093/jac/12.suppl_c.51. [DOI] [PubMed] [Google Scholar]
  17. Zimmerli W., Lew P. D., Cohen H. J., Waldvogel F. A. Comparative superoxide-generating system of granulocytes from blood and peritoneal exudates. Infect Immun. 1984 Dec;46(3):625–630. doi: 10.1128/iai.46.3.625-630.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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