Antibiotic — Neutrophil Interactions Studied by Phase Partitioning

  • Paul Eggleton
  • Derek Fisher
  • Neville Crawford


In an investigation of phase-forming polymers concerning their stimulatory effect on neutrophils (PMNLs), a number of dextrans were found to stimulate neutrophil oxidative metabolism as detected by nitroblue tetrazolium (NBT) reduction [1]. This effect was removed by addition of polymyxin B sulphate — a cyclic polypeptide antibiotic which binds to lipid A [2] and neutralises many of the biological effects of bacterial endotoxin [3,4]. This indicated that contaminating bacterial lipopolysaccharide (LPS) is present in some batches of dextran, but can be removed by addition of polymyxin B. Unfortunately, we observed that polymyxin B also enhanced the phagocytic capacity of the neutrophil. Consequently its incorporation into the phase reagents as a means of neutralising the effects of LPS is precluded. These observations prompted us to examine the surface properties of neutrophils in the presence and absence of polymyxin B by single step partitioning in charge-sensitive and non-charge-sensitive phase systems and the contribution of these surface properties to the observed increase in neutrophil phagocytosis.


Partition Coefficient Acridine Orange Bacterial Lipopolysaccharide Phagocytic Capacity Phase Partitioning 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    P. Eggleton, D. Fisher and N. Crawford, Problems of white cell separation techniques in relation to phase partitioning of neutrophils, in: “Separations Using Aqueous Phase Systems: Applications in Cell Biology and Biotechnology,” D. Fisher and I.A. Sutherland, eds Plenum, New York (1989)Google Scholar
  2. 2.
    D.C. Morrison and D.M. Jacobs, Binding of polymyxin B to the lipid A portion of bacterial lipopolysaccharides, Immunochemistry 13:813 (1976)PubMedCrossRefGoogle Scholar
  3. 3.
    J.J. Corrigan and B.M. Bell, Endotoxin induced intravascular coagulation; prevention with polymyxin B sulphate, J. Lab. Clin. Med 77:802 (1971)PubMedGoogle Scholar
  4. 4.
    J.J. Corrigan, O.F. Sieber, H. Ratajczak, and B.B. Bennet, Modification of human neutrophil response to endotoxin with polymyxin B sulphate, J. Infect. Dis. 130:384 (1974)PubMedCrossRefGoogle Scholar
  5. 5.
    C.M. Kunin, Binding of antibodies to tissue homogenates, J. Infect. Dis. 121:55 (1970)PubMedCrossRefGoogle Scholar
  6. 6.
    P.H. Naccache, M.M. Molski and R.I. Sha’afi, Polymyxin B inhibits phorbol 12-myristate 13-ocetate, but not chemotactic factor induced effects in rabbit neutrophils, FEBS 193:227 (1985)CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Paul Eggleton
    • 1
  • Derek Fisher
    • 1
  • Neville Crawford
    • 2
  1. 1.Biochemistry Department, Royal Free Hospital School of MedicineUniversity of LondonLondonUK
  2. 2.Biochemistry DepartmentRoyal College of Surgeons of EnglandLondonUK

Personalised recommendations