Testing Anti-Pneumococcal Antibody Function Using Bacteria and Primary Neutrophils

Methods Mol Biol. 2021:2183:559-574. doi: 10.1007/978-1-0716-0795-4_33.

Abstract

Antibodies against Streptococcus pneumoniae (pneumococcus) following vaccination are crucial for host protection against invasive pneumococcal infections. The antibodies induced by pneumococcal vaccines act as opsonins to mediate bacterial uptake and killing by host phagocytic cells, especially polymorphonuclear leukocytes (PMNs) also called neutrophils. Therefore, it is important to measure not only the levels of antibodies induced by a pneumococcal vaccine candidate but their actual functional capacity in mediating bacterial opsonization and killing by PMNs. Here, we describe a protocol to demonstrate effective deposition of vaccine-induced antibodies on the surface of S. pneumoniae by flow cytometry and subsequent opsonophagocytic killing (OPH) by murine bone-marrow derived PMNs.

Keywords: Antibodies; Flow cytometry; Neutrophils; Opsonophagocytic killing; Streptococcus pneumoniae.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Bacterial / immunology*
  • Biomarkers
  • Bone Marrow Cells / immunology
  • Bone Marrow Cells / metabolism
  • Flow Cytometry
  • Immune Sera / immunology
  • Mice
  • Neutrophils / immunology*
  • Neutrophils / metabolism
  • Phagocytes / immunology
  • Phagocytes / metabolism
  • Pneumococcal Infections / immunology*
  • Pneumococcal Infections / metabolism
  • Pneumococcal Infections / microbiology
  • Pneumococcal Vaccines / immunology*
  • Streptococcus pneumoniae / immunology*

Substances

  • Antibodies, Bacterial
  • Biomarkers
  • Immune Sera
  • Pneumococcal Vaccines