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Proliferation of MOG^35-55-specific T cell receptor lymphocytes from transgenic mice in response to autoantigen in vivo and in vitro
Location: 93
Mentor: Dr. Robert Levy
Multiple sclerosis (MS) is a chronic neurodegenerative autoimmune disease characterized by central nervous system inflammation and demyelination. Experimental autoimmune encephalomyelitis (EAE) is an experimental animal model of MS, induced by exposing the recipient immune system to myelin. A TCR transgenic mouse (2D2) expressing CD4+ T-lymphocytes with V𝛼3.2+/V𝛽11+ T cell receptors specific for myelin oligodendrocyte glycoprotein (MOG35-55) provides a valuable tool to study the underlying mechanisms controlling EAE progression. The primary objective here was to demonstrate the specificity of the 2D2 TCR for MOG35-55 autoantigen to assess antigen specific proliferation kinetics in response to peptide stimulation in vivo and in vitro. Lymphocytes were plated with varying doses of MOG35-55 peptide to identify optimal concentrations. Proliferation was assessed via cell counts (hemocytometer), and phenotypic analysis was performed via microscopy. The strongest response regarding cell proliferation following MOG35-55 stimulation in vitro was found at 96 hours. This assay confirmed specificity of the 2D2 TCR for the myelin peptide and defined proliferation kinetics in response to autoantigen. Following adoptive transfer of 2D2 T cells to normal mice and EAE induction, 2D2 T cell proliferation was assessed by Ki67 staining utilizing flow cytometry. A shift to effector phenotype demonstrated functional responsiveness against myelin in vivo, and we found a decline in proliferative capacity of 2D2 T cells in the blood over time. We posit this correlates with their presence in the spinal cord of animals with disease. Further investigations of 2D2 T cells driving EAE pathophysiology in the central nervous system are ongoing.
