Project description:To investigate the molecular pathways altered during aging of the lacrimal gland in C57BL/6J females, we compared the LG transcriptome at different time points.
Project description:Aging is associated with a massive infiltration of T lymphocytes in the lacrimal gland. . Among the non-Tregs, we also found a significant increase in the levels of EOMESmed/high, TbetnegIFN- + and CD62L+CD44neg CD4+ T cells with aging, which are associated with cell exhaustion, immunopathology and the generation of tertiary lymphoid tissue. More importantly, human lacrimal glands (age range 55-81 years) also showed presence of CD4+Foxp3+ cells. Further studies are needed to propose potential molecular targets to avoid immune-mediated lacrimal gland dysfunction with aging. Here we aimed to characterize the immune phenotype of aged CD4+ T cells in this tissue as compared with lymphoid organs. To perform this, we sorted regulatory T cells (Tregs, CD4+CD25+GITR+) and non-Tregs (CD4+CD25negGITRneg) cells and subjected these cells to an immunology NanoStringⓇ panel in lymphoid organs. These results were confirmed by flow cytometry, live imaging and tissue immunostaining in the lacrimal gland. Importantly, effector T helper 1 (Th1) genes were highly upregulated on aged Tregs, including the master regulator Tbx21
Project description:The purpose of the present studies was to use CyTOF and RNA-Seq technologies to identify cells and genes involved in lacrimal gland repair that could be targeted to treat diseases of lacrimal gland dysfunction. Lacrimal glands of female BALB/c mice were experimentally injured by intra-glandular injection of interleukin 1 alpha (IL-1α). The lacrimal glands were harvested at various time points following injury (1 to 14 days) and used to either prepare single cell suspensions for CyTOF immuno-phenotyping analyses or to extract RNA for gene expression studies using RNA-Seq. CyTOF immuno-phenotyping identified monocytes and neutrophils as the major infiltrating populations 1 and 2 days post injury. Clustering of significantly differentially expressed genes identified 13 distinct molecular signatures: 3 associated with immune/inflammatory processes included genes up-regulated at days 1-2 and 3 associated with reparative processes with genes up-regulated primarily between days 4 and 5. Finally, clustering identified 65 genes which were specifically up-regulated 2 days post injury which was enriched for muscle specific genes. The expression of select muscle-related proteins was confirmed by immunohistochemistry which identified a subset of cells expressing these proteins. Double staining experiments showed that these cells are distinct from the myoepithelial cells. We conclude that experimentally induced injury to the lacrimal gland leads to massive infiltration by neutrophils and monocytes which resolved after 3 days. RNAseq and immunohistochemistry identified a group of cells, other than myoepithelial cells, that express muscle-related proteins that could play an important role in lacrimal gland repair.
Project description:Our objective was to determine the nature and extent of androgen regulation of gene expression in the female lacrimal, meibomian,and submandibular glands, and to explore the degree to which this control is the same as in male glands. Keywords: Hormone treatment
Project description:Series includes pooled (n = 5 mice per biological replicate) samples from lacrimal, meibomian, and submandibular glands of male palcebo- and testosterone-treated BALB/c mice. All experiments were run in triplicate (pooled biological replicates) on CodeLink Mouse Uniset I Microarrays. Keywords: repeat sample
Project description:Prior to the onset of autoimmune destruction, type 1 diabetic patients and an animal model thereof, the nonobese diabetic (NOD) mouse, show morphological and functional abnormalities in target organs, which may act as inciting events for leukocyte infiltration. To better understand these abnormalities, but without the complications associated with inflammatory infiltrates, we examined genes expressed in autoimmune target tissues (pancreas, submandibular glands, and lacrimal glands) of NOD/scid mice and of autoimmune-resistant C57BL6/scid mice. Keywords: tissue expression, disease prone versus resistant strain comparison
Project description:Series includes pooled (n = 5 mice per biological replicate) samples from submandibular, sublingual, parotid, lacrimal, and meibomian glands of BALB/c mice. Both male and female samples were analyzed on CodeLink Mouse Uniset I Microarrays. Keywords: repeat sample
Project description:NOD mice were injected once a week with LTBR-Ig to block the LTBR-pathway, or with control monoclonal antibody MOPC from age 8 to 16 weeks old. Extraorbital lacrimal glands or submaxillary glands were dissected and total mRNA prepared. Each sample was either the combined lacrimals (2) from each mouse or individual salivary glands. There were 4 mice in each treatment group. Total mRNA was isolated and the quality was assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA). Reverse transcription to prepare cDNA was performed using Invitrogen M-MLV system. The purpose was to determine changes in gene expression in glands due to blockade of the LTBR-pathway. Differential Gene Expression in NOD mouse lacrimal and salivary glands after LTBR-Ig treatment
Project description:We used freshly isolated lacrimal glands isolated from wild type and Aire-/- mice on a Balbc background. Mice were 5 weeks of age, with an n=4 for wild type and n=3 for Aire-deficient.