Abstract
BackgroundUtilizing mouse models provides excellent immunological and experimental tools to study oral immune responses. However for functional assays, isolating T lymphocytes from the oral tissues has proved to be challenging due to the absence of reliable methods that yield viable cells with consistency. To study adaptive immune cell interactions in the oral mucosal tissues, it is necessary to isolate T cells with a good viability and study them at the single cell level.FindingsWe have established an improved method to isolate immune cells, including Tregs and Th17 cells from intra-epithelial niches and lamina propria of the tongue, gingival and palatal tissues in the oral mucosa of mice.ConclusionThis new method of isolating immune cells from oral tissues will enable us to further our understanding of oral tissue immune cells and their role during oral infections and oral inflammation.
Highlights
Utilizing mouse models provides excellent immunological and experimental tools to study oral immune responses
Oral immunity and inflammation are associated with a wide array of human health conditions including, but not limited to, cancer, cardiovascular disease, graft versus host disease (GVHD), and infectious diseases such as AIDS [1,2,3,4,5]
Studies have focused on T cells which are present in intra-epithelial compartments; i.e., oral lymphoid foci, and lamina propria [8,9,10,17], and determined that these cells play significant roles in oral immunity and inflammation [7,11,12]
Summary
Utilizing mouse models provides excellent immunological and experimental tools to study oral immune responses. T cells have been studied in oral tissues by flow cytometry [13], the cells isolated using such crude methods frequently show poor viability. The epithelial cells and intra-epithelial lymphocytes within the sedimented tissues are further removed by intense vortexing in 5 ml of RPMI-1640 containing antibiotics, 2 mM EDTA and 20 mM HEPES.
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