Lacrimal gland epithelial cells stimulate proliferation in autologous lymphocyte preparations

Autoimmune dacryoadenitis is a frequent cause of lacrimal insufficiency. In order to test hypotheses regarding mechanisms that can trigger this syndrome, we developed a method to obtain a preparation of rabbit lacrimal gland epithelial cells essentially free of immune-system cells. The method relies on controlled digestion to disperse lacrimal acini, and recovers acini by filtration through various sizes of nylon mesh. Purity and integrity of the preparation were established qualitatively using light and electron microscopy. Contamination by immune-system cells was quantitated by immunohistochemistry using anti-CD18, and -RTLA (rabbit thymic lymphocyte antigen) antibodies. The novel method produced preparations of highly-purified lacrimal gland epithelial cells (pLGEC) with expected morphological characteristics with less than 1.5% of the cells staining for CD18 or RTLA. The method also yielded preparations of lacrimal gland interstitial cells (LGIC) enriched for lymphocytes; in these preparations either CD18 or RTLA were detected on nearly 10% of the cells. pLGEC promoted proliferation in preparations of autologous splenic lymphocytes (SPL) that was blocked by anti-MHC class II but not anti-MHC class I antibodies. This observation, combined with the apparent requirement that pLGEC must contact the autologous lymphocyte preparation to promote proliferation, supports the hypothesis the proliferation arises from antigen-presentation via MHC class II by pLGEC.