Precision targeting of autoantigen-specific B cells in muscle-specific tyrosine kinase myasthenia gravis with chimeric autoantibody receptor T cells - Nature Biotechnology

. pMSGV1.MuSK-CAAR was generated by replacing the 1D3-28Z.1-3 mut insert with the MuSK-CAAR sequence. Then 30 μg of each plasmid was transfected into the Plat-E packaging cell line to produce retroviruses.MG3, a 57-year-old female, chronic active disease. MG5, a 34-year-old female, chronic active disease. Venipuncture was performed under a protocol approved by the University of Pennsylvania Institutional Review Board.

Bulk primary human T cells ) were cultured in human T cell culture media supplemented with 100 IU mlrhIL-2 plus 5% human AB serum or Roswell Park Memorial Institute media supplemented with 10% FBS, 10 mM HEPES, 1% penicillin/streptomycin and 1% GlutaMax). T cells were activated/selected with anti-CD3/CD28-coated paramagnetic beads at a 3/1 bead/cell ratio.

Donor-matched MuSK-CAART and NTD-T were incubated with BCR-negative Nalm-6 control cells or mixture of Nalm-6 MuSK target cells at E:T ratio of either 1:1 or 10:1 for 24 h. Purified IgG from two patients with MuSK MG was added at a final concentration of 10 mg mlIgG before coincubation. MuSK-CAART cytotoxicity was evaluated at 8 and 24 h using luciferase-based killing assay.

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PennMedicine CabalettaBio UCDavisHealth NeurologyYale Targeting autoimmune B-cells with chimeric autoantibody receptors (CAARs): behindthepaper with co-authors Silvio Manfredo-Vieira and Aimee S. Payne

PennMedicine CabalettaBio UCDavisHealth NeurologyYale CAR T-like treatment for rare form of autoimmune disease NBTintheNews via PennMedicine

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