A Murine Model of Primed Mycobacterial Uveitis to Study Post-Infectious Uveitis
Published: September 29, 2023
Abstract
Source: John, S., et al. Primed Mycobacterial Uveitis (PMU) as a Model for Post-Infectious Uveitis. J. Vis. Exp. (2021).
This video demonstrates a method to generate a mouse model of primed mycobacterial uveitis (PMU) for studying post-infectious uveitis. Upon induction of PMU by intravitreal injection of heat-killed Mycobacterium tuberculosis in a mouse previously immunized with the same immunogen, the progression of chronic inflammation in the eye is analyzed using imaging and histopathological examination
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). Th…
Representative Results
Figure 1: Mouse intravitreal injection setup. The intravitreal injection is performed on the mouse eye using (A) a micro syringe pump controller connected to the (B) Micropump and (C) an injection syringe. The syringe is loaded and mounted on the (D) Micropump. The mouse head is positioned using (E) ear bars to e…
Disclosures
The authors have nothing to disclose.
Materials
| AK-FLUOR | Akorn Pharmaceuticals, IL, USA | 10% Fluorescein sodium 100 mg/mL in 5 mL vial | |
| AnaSed | Akorn Animal Health, IL, USA | NDC 59399-110-20 | Xylazine 20 mg/mL |
| Betadine 5% Sterile Ophthalmic Prep Solution | Alcon, TX, USA | 8007-1 | |
| B-D Precision Glide Needles -25 G | Becton, Dickinson and Company, NJ, USA | 305122 | |
| B-D Precision Glide needle -30-G | Becton, Dickinson and Company, NJ, USA | 305106 | |
| Bond MAX, Bond Rx | Leica Biosystems, IL,USA | Automated IHC staining system | |
| Chloramphenicol ointment | Martindale Pharma, Romford, UK | 1% w/w Chloramphenicol | |
| EG1150H | Leica Biosystems, IL,USA | Tissue Embedding | |
| Envisu R2300 | Bioptigen/Leica | OCT Machine | |
| Freund's Incomplete Adjuvant | BD Difco, NJ, USA | 263910 | |
| GenTeal lubricant eye ointment | Alcon, TX, USA | — | |
| GenTeal lubricant eye gel | Alcon, TX, USA | — | |
| H37Ra lyophilized Mycobacteria extract | BD Difco, NJ, USA | 231141 | |
| Hamilton RN Needle (33/12/2)S | Hamilton, Reno, NV | 7803-05(33/12/2) | 33 G |
| Hamilton syringe | Hamilton, Reno, NV | CAL7633-01 | 5 µL |
| Insulin needle | Exel International, USA | 26029 | 1 mL |
| Isoflurane | |||
| Ketaset | Zoetis, USA | 377341 | Ketamine HCL 100 mg/mL |
| Microinjection Syringe Pump and Micro4Controller | World Precision Instruments, FL, USA | UMP3 | |
| Micron IV | Phoenix Research Laboratories, Pleasanton, CA | Alternative Imaging/OCT Machine | |
| Nanofil 10 µL syringe | World Precision Instruments, FL, USA | NANOFIL | |
| Nanofil Intraocular Injection Kit | World Precision Instruments, FL, USA | IO-KIT | |
| Olympus SZX10 | Olympus | Dissection scope | |
| PBS | Gibco | 14190 | |
| Phenylephrine Hydrochloride Ophthalmic Solution USP 2.5% Sterile 15 mL | Akorn Pharmaceuticals, IL, USA | 17478020115 | |
| RM2255 | Leica Biosystems, IL,USA | Tissue Sectioning | |
| TB Syringe | Becton, Dickinson and Company, NJ, USA | 309602 | 1 mL |
| Tetracaine 0.5% | Alcon, TX, USA | 1041544 | |
| Tissue Tek VIP series | Sakura Finetek USA, Inc.,CA. | Histology Tissue Processing | |
| Tropicamide 1% | Chauvin Pharmaceuticals, Romford, UK | Minims | |
| Tylenol | Johnson & Johnson Consumer Inc, PA, USA | NDC 50580-614-01 | Acetaminophen |
| Viscotears | Novartis Pharmaceuticals, Camberley, UK | Carbomer eye gel 0.2% w/w |
Tags
Cite This Article
A Murine Model of Primed Mycobacterial Uveitis to Study Post-Infectious Uveitis. J. Vis. Exp. (Pending Publication), e21704, doi: (2023).