Establishment of Orthotopic Lacrimal Gland B-cell Lymphoma Model in Mice

Abstract

Purpose : To develop and characterize a lacrimal gland B-cell lymphoma model in mice.

Methods : Either 1x10⁵, or 1x10⁶ syngeneic lymphomatous cell line (A20 cell line) suspended in 20ul phosphate buffer solution were injected into intraorbital and extraorbital lacrimal glands. Lacrimal glands were dissected and the volume was measured at 1, 2, 3, and 4 weeks after injection. Infiltration of tumor cells and damage to the lacrimal glands were evaluated by histopathology. Also, the cellular components of tumor microenvironment were studied by immunofluorescence staining.

Results : The volume of intraorbital lacrimal glands was significantly larger in 1x10⁶ A20 group at 1 week after injection (p=0.0117 vs no injection group, p=0.0081 vs 1x10⁵ A20 group). The volume of extraorbital lacrimal was also significantly increased in 1x10⁶ A20 group at 1 week (p=0.009 vs no injection group, p=0.0040 vs 1x10⁵ A20 group) and 2 week (p=0.0044 vs no injection group, p=0.0283 vs 1x10⁵ A20 group) after injection. The volume of lacrimal glands was not significantly different among three groups 3 weeks and 4 weeks after injection. Tumor cells intensively invaded and diffusely observed in whole intraorbital and extraorbital lacrimal gland specimens obtained at 1 and 2 weeks after injection. Lacrimal glands at 3 weeks and 4 weeks showed only focal tumor cell infiltration. CD19+ B cells were abundantly infiltrated in lacrimal glands at 1 week and 2 weeks. CD3+ T cells and CD11b+ myeloid-derived suppressor cells were accompanied with tumor.

Conclusions : We established lacrimal gland B-cell lymphoma model in mice by orthotopic injection of A20 cells. The tumorigenesis effect was greater in high dose injection group, and the time course peaked 1 week after injection in intraorbital glands, and 2 weeks in extraorbital glands.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

 

The volume of intraorbital lacrimal glands was significantly higher in 1x10⁶ A20 injection group at 1week after injection. In case of extraorbital lacrimal glands, the volume was greater in 1x10⁶ A20 grou at 1 week and 2 weeks after injection.