This video showcases a cell-based therapy for autoimmune arthritis in mice. iPSC-derived T regulatory cells expressing ovalbumin-specific receptors are injected with methylated bovine serum albumin (mBSA) emulsified in an adjuvant. Murine antigen-presenting cells (APCs) process and present the mBSA to T cells, prompting T cell activation and migration to the knee joint's synovial membrane. Subsequent injections involve mBSA alone in the left knee and a combination of mBSA and ovalbumin in the right knee. In the left knee, the immune response to mBSA triggers arthritis, while in the right knee, T-regulatory cells interacting with ovalbumin mitigate inflammation, improving arthritis symptoms.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. In Vivo Maturation and Suppression of Autoimmune Arthritis Generate the constructs to be used in retroviral transduction by sub-cloning the OT-II T cell receptor (TCR) genes and forkhead box protein 3 (FoxP3) into the MiDR plasmid linked with self-cleaving peptide 2A to make the MiDR-TCRα-2A…
Representative Results
Figure 1: Adoptive Transfer of Ag-specific iPSC-Tregs Ameliorates AIA in Mice. Murine iPSCs were transduced with the retroviral construct MiDR, MiDR-FoxP3, or MiDR-TCR-FoxP3 and were co-cultured on the OP9-DL1/DL4/I-Ab cells. On day 7, the gene-transduced cells (3 × 106/mouse) were adoptively transferred into female C57BL/6 mice induced with AIA two wee…