Source: Haque, M. et al., Development of Stem Cell-derived Antigen-specific Regulatory T Cells Against Autoimmunity. J. Vis. Exp. (2016)
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.
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
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 weeks after the cell transfer. On the day following arthritis induction, the arthritis severity was monitored by measurement of the knee diameter. (A-C) Percent increase in knee diameter. An increase in knee diameter was calculated based on preinjection knee diameter for each mouse before injection on day 0. Arthritis score was evaluated by examining both knees in a blinded manner; each knee was assigned a score (0: no visible swelling or discoloration; 1: visible swelling with or without discoloration; 2: moderate swelling with discoloration; 3: severe swelling with discoloration). In each group, five mice were used, and the data are representative of three independent experiments. Data are represented as the mean ± SD. (D) The mean scoring on day 7 for both knees was from five mice. Data are represented as the mean ± SD from three independent experiments (** p< 0.01, *** p< 0.001, two-way ANOVA).
The authors have nothing to disclose.
C57BL/6j mice | Jackson Laboratory | 664 | |
α-MEM | Invitrogen | A10490-01 | |
FBS | Hyclone | SH3007.01 | |
Polybrene | Sigma | 107689 | |
Genejammer | Integrated science | 204130 | |
mFlt-3L | peprotech | 250-31L | |
mIL-7 | peprotech | 217-17 | |
mBSA | Sigma | A7906 | |
Ova albumin | Avantor | 0440-01 | |
CFA | Difco | 2017014 | |
Tailveiner restrainer | Braintree scientific | RTV 150-STD | |
Paraformaldehyde | Sigma | P6148-500G | Caution: Allergenic, Carcenogenic, Toxic |
.