Determination of Active Phagocytosis of Unopsonized Porphyromonas gingivalis by Macrophages and Neutrophils Using the pH-Sensitive Fluorescent Dye pHrodo

Infect Immun. 2016 May 24;84(6):1753-1760. doi: 10.1128/IAI.01482-15. Print 2016 Jun.

Abstract

Phagocytosis of pathogens is an important component of the innate immune system that is responsible for the removal and degradation of bacteria as well as their presentation via the major histocompatibility complexes to the adaptive immune system. The periodontal pathogen Porphyromonas gingivalis exhibits strain heterogeneity, which may affect a phagocyte's ability to recognize and phagocytose the bacterium. In addition, P. gingivalis is reported to avoid phagocytosis by antibody and complement degradation and by invading phagocytic cells. Previous studies examining phagocytosis have been confounded by both the techniques employed and the potential of the bacteria to invade the cells. In this study, we used a novel, pH-sensitive dye, pHrodo, to label live P. gingivalis strains and examine unopsonized phagocytosis by murine macrophages and neutrophils and human monocytic cells. All host cells examined were able to recognize and phagocytose unopsonized P. gingivalis strains. Macrophages had a preference to phagocytose P. gingivalis strain ATCC 33277 over other strains and clinical isolates in the study, whereas neutrophils favored P. gingivalis W50, ATCC 33277, and one clinical isolate over the other strains. This study revealed that all P. gingivalis strains were capable of being phagocytosed without prior opsonization with antibody or complement.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial
  • Cell Line
  • Fluorescent Dyes / chemistry*
  • Host Specificity
  • Humans
  • Hydrogen-Ion Concentration
  • Immunity, Innate
  • Macrophages / immunology*
  • Macrophages / microbiology
  • Mice
  • Neutrophils / immunology*
  • Neutrophils / microbiology
  • Opsonin Proteins
  • Phagocytosis*
  • Porphyromonas gingivalis / growth & development
  • Porphyromonas gingivalis / immunology*
  • Spectrometry, Fluorescence
  • Staining and Labeling / methods

Substances

  • Antibodies, Bacterial
  • Fluorescent Dyes
  • Opsonin Proteins

Grants and funding

This work was funded by the Australian Government Department of Industry, Innovation and Science (Cooperative Research Centre grant 20080108) and a National Health and Medical Research Council project grant (APP1029878).