Subcutaneous Injection of Bacterial Antigen to Induce Systemic Inflammation in a Murine Model

Abstract

Source: John, S. et al., Primed Mycobacterial Uveitis (PMU) as a Model for Post-Infectious Uveitis. J. Vis. Exp. (2021)

This video demonstrates the induction of systemic inflammation in an anesthetized mouse. The injection of mycobacterial antigen-adjuvant depots triggers cytokine release and localized inflammation. Lymphatic vessels open, enabling immune cell entry into lymph nodes and activation of T cells, which release pro-inflammatory cytokines, leading to widespread inflammation.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Antigen preparation for subcutaneous injection Perform all procedures in this section inside a chemical fume hood to prevent inhalation or skin contact with the Mycobacterium tuberculosis, Mtb H37Ra powder. Handle according to your institutional policies for Complete Freund's Adjuvant (typically BSL-1). Thi…

Materials

Freund's Incomplete Adjuvant	BD Difco, NJ, USA	263910
H37Ra lyophilized Mycobacteria extract	BD Difco, NJ, USA	231141
Insulin needle	Exel International, USA	26029	1 mL
Isoflurane
PBS	Gibco	14190
RM2255	Leica Biosystems, IL,USA		Tissue Sectioning
TB Syringe	Becton, Dickinson and Company, NJ, USA	309602	1 mL
Tylenol	Johnson & Johnson Consumer Inc, PA, USA	NDC 50580-614-01	Acetaminophen