Home › Immunology › A Chronic Autoimmune Dry Eye Rat Model with Increase in Effector Memory T Cells in Eyeball Tissue
Immunology JoVE (Open Access) Citable · DOI

A Chronic Autoimmune Dry Eye Rat Model with Increase in Effector Memory T Cells in Eyeball Tissue

DOI: 10.3791/55592-v
What you'll learn
  • ✓ Prepare lacrimal gland extract and complete Freund's adjuvant emulsion for immunization
  • ✓ Induce chronic autoimmune dry eye via immunization and antigen injection in Lewis rats
  • ✓ Assess dry eye features including tear production and ocular surface damage
Protocol

Biopharma Insights This report describes a method to induce chronic experimental autoimmune dry eye in Lewis rats through immunization with an emulsion of rat lacrimal gland extract, ovalbumin, and complete Freund's adjuvant, followed by the injection of lacrimal gland extract and ovalbumin into the forniceal subconjunctiva and lacrimal glands six weeks later.

Difficulty
advanced
Total time
~8 weeks (immunization + 6-week latency + assessment period)
Model organism
Rat Lewis
Biosafety
BSL-1

Steps

1
Prepare lacrimal gland extract from donor rats

Extract lacrimal glands from donor Lewis rats and process tissue into a usable antigen extract. This extract serves as the primary immunogen for inducing autoimmune dry eye.

▶ 01:18
2
Prepare immunization emulsion with complete Freund's adjuvant

Combine lacrimal gland extract, ovalbumin, and complete Freund's adjuvant to create an emulsion suitable for subcutaneous immunization of Lewis rats.

▶ 02:32
3
Immunize rats and inject antigen into ocular tissues

Perform initial subcutaneous immunization with the emulsion, then inject lacrimal gland extract and ovalbumin directly into the forniceal subconjunctiva and lacrimal glands six weeks later to elicit chronic autoimmune dry eye.

▶ 03:30
4
Assess tear production and ocular surface damage

Perform clinical and histological assessments to measure dry eye features, including tear secretion, corneal erosion, and immune cell infiltration in eyeball tissue.

▶ 04:53
5
Analyze effector memory T cell populations in eyes

Characterize effector memory T cells and other immune markers in lacrimal gland and ocular tissues to confirm the autoimmune etiology of the dry eye model.

▶ 07:19
💬 Comments coming soon