Streptococcus pneumoniae

Growth and preparation of Streptococcus pneumoniae

Intranasal colonization of Streptococcus pneumoniae

  • This is a detailed protocol for preparing Streptococcus pneumoniae and intranasal colonization of mice.

CFSE labelling of S pneumoniae

  • This protocol uses CFSE to label S. pneumoniae for flow cytometry or microscopy experiments.

NF-KB Luciferase Assay using HEK293T cells

  • This protocol can be used to test whether bacterial ligands induce activation of TLRs and NLRs.

Macrophage Killing Assay version 1

Macrophage Killing Assay version 2

  • How do you measure how well macrophages kill bacteria (or whether your treatment alters killing)? This is a standard kill curve protocol and a workhorse of the Bowdish lab. Specifically written to measure S. pneumoniae killing, can be adapted for any bacteria and any type of macrophage (e.g. human, murine, alveolar, bone marrow,etc).

Pneumococcal Cell Wall Purification

  • This protocol describes how to purify the pneumococcal cell wall. The purified preparation will not contain proteins but will contain the peptidoglycan layer and teichoic acids. It is updated from the original.

Phenol-sulfuric acid Mannose detection assay.

  • This assay is sensitive to mannose from 1 – 100 𝜇g but will also detect other carbohydrates. We use this assay to quantify of complex carbohydrates in the bacterial cell wall
 Nasopharyngeal colonization, and measurement of RNA and leukocyte recruitment.
For a detailed protocol, see our paper here…

Puchta A, Verschoor CP, Thurn T, Bowdish DM. Characterization of inflammatory responses during intranasal colonization with Streptococcus pneumoniae. J Vis Exp. 2014 Jan 17;(83):e50490. doi: 10.3791/50490.

Leave a Reply