A Non-Human Primate Model of Acute Group A Streptococcus Pharyngitis - Bacterial Pathogenesis: Methods and Protocols - page 242

This chapter describes a non-human primate model of GAS pharyngitis that
we have used successfully to gain insight into the transcriptional adaptation of
GAS during upper respiratory tract infection (21,22) , to determine the contri-
bution of specific GAS virulence factors to infection (23,24) , and to investigate
possible vaccine candidates (unpublished data). The following protocol outlines
a comparison study where wild-type and isogenic mutant strains of GAS are used
to determine the contribution of a putative virulence gene to GAS pharyngitis.
2. Materials
2.1. Preparation of GAS inoculum
1. Wild-type and mutant GAS strains.
2. Todd-Hewitt broth (THY broth) with 0.2% yeast extract.
3. 80% glycerol, sterile.
4. THY agar plates (THY broth plus 1.5% agar).
5. Phosphate-buffered saline (PBS), sterile.
6. 50-mL falcon tubes, sterile.
7. 15-mL screw cap tubes, sterile.
8. Liquid nitrogen.
2.2. Animal Grouping
1. Purified streptolysin-O-protein (SLO, Sigma cat no. S5265).
2. Bovine serum albumin (BSA).
3. PBS with 0.1% Tween 20 (PBST).
4. Goat-anti-monkey IgG-HRP.
5. 3,3´,5,5´-tetramethylbenzidine solution (TMB).
6. 0.18 M sulfuric acid (stop solution).
7. 96-well clear bottom ELISA plates.
2.3. Non-human Primate Infections
1. Eight cynomolgus macaques ( Macaca fascicularis ).
2. 1-mL syringes.
3. PBS, sterile.
2.4. Disease Development
1. Laryngoscope (reusable handle with disposable blades).
2.5. Blood Draws
1. Eight Vacutainer™ systems (VWR).
2. Red-capped Vacutainer™ plasma tubes (VWR).
3. 2-mL red-capped tubes, sterile.