Promotion of opsonization by antibodies and phagocytosis of Gram-positive bacteria by a bifunctional polyacrylamide

Biomaterials. 2006 Jul;27(19):3663-74. doi: 10.1016/j.biomaterials.2006.02.006. Epub 2006 Mar 9.

Abstract

This paper describes the application of a bifunctional polyacrylamide (pA-V-F) presenting both vancomycin and fluorescein groups, to modify the surfaces of multiple species of Gram-positive bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, and Enterococcus faecalis) to control molecular recognition of these surfaces. The vancomycin groups allowed the specific recognition of a structural component of the bacterial cell wall: peptides terminated in D-Ala-D-Ala. The fluorescein groups allowed the imaging of binding of polymer to the surfaces of bacteria by fluorescence, and are representative, low molecular weight haptens; their recognition by anti-fluorescein antibodies provides proof-of-principle that bifunctional polymers can be used to introduce haptens onto the surface of the bacteria. Flow cytometry revealed that polymer-labeled S. aureus and S. pneumoniae were opsonized by anti-fluorescein antibodies approximately 20-fold more than were untreated bacteria; nearly all ( approximately 92%) polymer-labeled S. aureus, and a large (76%) fraction of polymer-labeled S. pneumoniae were opsonized. The bound antibodies then promoted phagocytosis of the bacteria by cultured J774 macrophage-like cells. Flow cytometry revealed that macrophages ingested S. aureus decorated with the polymer-antibody complexes approximately 2-fold more efficiently than S. aureus in control groups, in spite of the high background (caused by efficient antibody-independent ingestion of S. aureus by macrophages). This paper, thus, demonstrates the ability of a bifunctional polymer to carry out three distinct functions based on polyvalent molecular recognition: (i) recognition of the surface of Gram-positive bacteria, (ii) modification of this surface to generate specific binding sites recognized by an antibody, and (iii) promotion of phagocytosis of the opsonized bacteria.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acrylic Resins / chemistry
  • Acrylic Resins / pharmacology*
  • Animals
  • Antibodies, Bacterial / metabolism*
  • Biocompatible Materials / chemistry
  • Biocompatible Materials / pharmacology*
  • Cell Line
  • Enterococcus faecalis / drug effects
  • Enterococcus faecalis / immunology
  • Gram-Positive Bacteria / drug effects*
  • Gram-Positive Bacteria / immunology*
  • In Vitro Techniques
  • Macrophages / drug effects
  • Macrophages / immunology
  • Materials Testing
  • Mice
  • Microscopy, Fluorescence
  • Opsonin Proteins / metabolism*
  • Phagocytosis / drug effects
  • Staphylococcus aureus / drug effects
  • Staphylococcus aureus / immunology
  • Staphylococcus epidermidis / drug effects
  • Staphylococcus epidermidis / immunology

Substances

  • Acrylic Resins
  • Antibodies, Bacterial
  • Biocompatible Materials
  • Opsonin Proteins
  • polyacrylamide