Class
Article
College
College of Agriculture and Applied Sciences
Department
Animal, Dairy, and Veterinary Sciences Department
Presentation Type
Oral Presentation
Abstract
Embryonic loss is an important problem in the cattle industry. Therefore, an understanding of mechanisms that regulate placental and embryonic development, such as the immune regulatory interactions that exist between placental cells and immune cells during pregnancy, is relevant for this industry. Our hypothesis is that trophoblast cells from functional placentas secrete extracellular-vesicles (EV) that can alter the gene expression profile of immune cells as a mechanism of immune regulation. To test this conjecture, bovine placental cells were isolated and cultured for 48h, conditioned media was harvested, and 50-1000 nm EVs were isolated by a differential ultracentrifugation method. Cells from ten animals were used to prepare the mix of EVs. Different immune cell populations (CD4+ CD25+ T lymphocytes, CD4+ CD25- T lymphocytes, CD8+ T lymphocytes, γ/δ T cells, and monocytes) were sorted by flow cytometry. These cells were either cultured alone (control group) or supplemented with trophoblast-derived EVs for 48 hours. Total cell RNA was isolated for gene expression analysis by microfluidic chip, real-time RT-PCR (96.96 Dynamic Array; Fluidigm). Gene expression analysis was conducted for the following cytokines, transcription factors and CD markers: IL1, IL2, IL4, IL5, IL6, IL8, IL10, IL12B, IL13, IL15, IL17, IL18, IL23, IFNG, TNF, TGFB1, CSF2, FoxP3, TBX21, GATA3, CD25, IL2RA, and CTLA4. A strong effect of trophoblast cell derived EV on the gene expression profile of immune cells was detected. The different cell populations analyzed responded differently to trophoblast derived EVs. Transcription factors GATA3 and FoxP3 were downregulated in both CD4+ populations, while they were upregulated in CD8+ cells. IL2 and IL17 were downregulated in CD4+CD25+ cells and upregulated in CD8+ cells treated with trophoblast-derived EVs. Both CD4+ cell populations downregulated TGFB1 expression when treated with EVs. The overall gene expression profile indicates that trophoblast-derived EVs regulate immune cell activity.
Location
Room 101
Start Date
4-11-2019 9:00 AM
End Date
4-11-2019 10:15 AM
Included in
Gene Expression Profiles of Immune Cells Under the Influence of Bovine Trophoblast Cell Derived Extracellular Vesicles
Room 101
Embryonic loss is an important problem in the cattle industry. Therefore, an understanding of mechanisms that regulate placental and embryonic development, such as the immune regulatory interactions that exist between placental cells and immune cells during pregnancy, is relevant for this industry. Our hypothesis is that trophoblast cells from functional placentas secrete extracellular-vesicles (EV) that can alter the gene expression profile of immune cells as a mechanism of immune regulation. To test this conjecture, bovine placental cells were isolated and cultured for 48h, conditioned media was harvested, and 50-1000 nm EVs were isolated by a differential ultracentrifugation method. Cells from ten animals were used to prepare the mix of EVs. Different immune cell populations (CD4+ CD25+ T lymphocytes, CD4+ CD25- T lymphocytes, CD8+ T lymphocytes, γ/δ T cells, and monocytes) were sorted by flow cytometry. These cells were either cultured alone (control group) or supplemented with trophoblast-derived EVs for 48 hours. Total cell RNA was isolated for gene expression analysis by microfluidic chip, real-time RT-PCR (96.96 Dynamic Array; Fluidigm). Gene expression analysis was conducted for the following cytokines, transcription factors and CD markers: IL1, IL2, IL4, IL5, IL6, IL8, IL10, IL12B, IL13, IL15, IL17, IL18, IL23, IFNG, TNF, TGFB1, CSF2, FoxP3, TBX21, GATA3, CD25, IL2RA, and CTLA4. A strong effect of trophoblast cell derived EV on the gene expression profile of immune cells was detected. The different cell populations analyzed responded differently to trophoblast derived EVs. Transcription factors GATA3 and FoxP3 were downregulated in both CD4+ populations, while they were upregulated in CD8+ cells. IL2 and IL17 were downregulated in CD4+CD25+ cells and upregulated in CD8+ cells treated with trophoblast-derived EVs. Both CD4+ cell populations downregulated TGFB1 expression when treated with EVs. The overall gene expression profile indicates that trophoblast-derived EVs regulate immune cell activity.